TECHNICAL FIELD
[0001] Generally, the present invention relates to the technical field of sample analysis,
such as the analysis of biological samples, and further to the technical field of
high throughput analysis of biological samples.
[0002] In particular, the present invention is directed to a device for thermocycling biological
samples, to an instrument for simultaneously monitoring multiple nucleic acid amplification
reactions during thermocycling biological samples by means of such device, and also
to a method for thermocycling biological samples using such device for thermocycling
biological samples.
[0003] In other words, the present invention relates to a thermocycling structure or device,
also referred to as thermocycler or thermal cycler, for performing chemical and/or
biological reactions, such as, for example, Polymerase Chain Reactions (PCR), wherein
such thermocycling device can be provided as internal part of a laboratory instrument,
and wherein such thermocycling device usually includes at least a sample mount, a
heat pump with a heat sink, as well as a control unit for controlling the heating
and cooling of the heat pump during thermocycling. The present invention also relates
to an instrument for simultaneously monitoring multiple nucleic acid amplification
reactions during thermocycling biological samples by means of such thermocycling device,
wherein the monitoring instrument further comprises an excitation light source for
applying excitation energy to the nucleic acid amplification reactions and a sensor
for simultaneously detecting light emitted from the multiple nucleic acid amplifications,
and moreover to a respective thermocycling method using such a thermocycling device.
BACKGROUND
[0004] Biological samples are usually taken from patients by medical personnel in hospitals
or in private practice, for laboratory analysis, e.g. for determining concentration
levels of different components within the taken samples. Accordingly, the terms "sample"
and "biological sample" refer to material(s) that may potentially contain an analyte
of interest, wherein the biological sample can be derived from any biological source,
such as a physiological fluid, including blood, saliva, ocular lens fluid, cerebrospinal
fluid, sweat, urine, stool, semen, milk, ascites fluid, mucous, synovial fluid, peritoneal
fluid, amniotic fluid, tissue, cultured cells, or the like, and wherein the sample
can be suspected to contain a certain antigen or nucleic acid.
[0005] For many biological, biochemical, diagnostic or therapeutic applications, it is essential
to be able to accurately determine the amount or concentration of a certain substance
or compound in a biological sample contained in a reaction mixture, such as a certain
antigen or nucleic acid as mentioned above. In order to be able to achieve this goal
accurately, methods have been developed over the years, such as the widely known Polymerase
Chain Reaction (PCR), for example in the form of a real-time PCR, digital PCR (dPCR)
or multiplex PCR, which enable the in vitro synthesis of nucleic acids in a biological
sample, through which a DNA segment can be specifically replicated, i.e. a cost-effective
way to copy or amplify small segments of DNA or RNA in the sample. The development
of these methods for amplifying DNA or RNA segments has generated enormous benefits
in gene analysis as well as the diagnosis of many genetic diseases, or also in the
detection of viral load. Usually, thermal cycling, also referred to as thermocycling,
can be utilized to provide heating and cooling of reactants in a sample provided inside
a reaction vessel for amplifying such DNA or RNA segments, wherein laboratory instruments
including thermocyclers are commonly used in order to achieve an automatic procedure
of diagnostic assays based on PCR, in which, during a PCR conduct, the liquid PCR-samples
have to be heated and cooled to differing temperature levels repeatedly and have to
be maintained for a certain amount of time at different temperature plateaus. In order
to be able to accurately maintain such temperature plateaus during thermal cycling,
thermal uniformity throughout a thermal block of a thermal cycler should be maintained,
so that different sample wells can be heated and cooled uniformly to obtain uniform
sample yields between samples wells.
[0006] In the course of a typical PCR conduct, a specific target nucleic acid is amplified
by a series of reiterations of a cycle of steps in which nucleic acids present in
the reaction mixture are (a) denatured at relatively high temperatures, for example
at a denaturation temperature of more than 90° C, usually about 94°-95°C, for separation
of the double-stranded DNA, then (b) the reaction mixture is cooled down to a temperature
at which short oligonucleotide primers bind to the single stranded target nucleic
acid, for example at an annealing temperature of about 52°-56° C for primer binding
at the separated DNA strands in order to provide templates (annealing), and, thereafter,
(c) the primers are extended/elongated using a polymerase enzyme, for example at an
extension temperature at about 72°C for creation of new DNA strands, so that the original
nucleic acid sequence has been replicated. Repeated cycles of denaturation, annealing
and extension, usually about 25 to 30 repeated cycles, result in the exponential increase
in the amount of target nucleic acid present in the sample, wherein the time for heating
and cooling the samples has a significant influence on the overall process time. Accordingly,
less time spent at non-optimum temperatures results in better or more precise chemical
outcomes. In particular, a specific minimum time for holding any reaction mixture
at each of the temperature plateaus is required after reaching the same, wherein such
minimum holding times the minimum time it takes to complete one thermal cycle. Any
time in transition between PCR temperature plateaus is time added to this minimum
cycle time. Therefore, since the number of thermal cycles can be large, such additional
time unnecessarily heightens the total time needed to complete PCR conduct. Thus,
a decrease in heating and cooling time is essential for an efficient and cost effective
process and an increase in throughput of a thermocycling device for PCR. Accordingly,
there is a need to make diagnostic assays faster, cheaper and simpler to perform while
achieving precision as well as efficiency.
[0007] Commonly known thermocycling devices for amplifying DNA segments by means of PCR
basically consist of a mount for receiving the samples, often also referred to as
a sample tempering mount, and a heat pump attached to the mount, wherein the combination
of mount and heat pump can also be referred to as heat block or thermal block. The
heat pump, often provided in the form of a thermoelectric device or thermoelectric
cooler (TEC), for example in the form of a Peltier element, is usually used for active
heating and cooling of the mount and, thus, for actively controlling the temperature
provided to the samples. TECs are solid-state heat pumps usually made from semiconductor
materials sandwiched between ceramic plates, wherein an amount of heat pumped is proportional
to the amount of current flowing through the TEC, resulting in increased temperature
control, wherein, by reversing the current, TECs can function as heaters or coolers,
which is highly useful for thermocycling at different temperatures. However, due to
the relatively large amount of heat being pumped over a small area, TECs in general
require some kind of heat dissipation means, such as a heat sink thermally coupled
to the TEC, to dissipate the heat away from the TEC and, for example, into the ambient
environment, which heat sinks are often additionally provided with a fan used as air
cooling means in order to facilitate heat dissipation of the heat sink to the ambient
air.
[0008] However, since the use of heat sinks combined with fans can causes significant noise
emission, and since such combinations for heat dissipation are usually heavy in weight,
alternative solutions have been developed in the recent past, for example the heating/cooling
of mounts by means of fluid carrying systems. As an example of such known prior art,
WO 2006/105919 A1 discloses a device for the simultaneous thermocycling of multiple samples, which
device basically comprises a mount for receiving the samples, a heat pump attached
to the mount, and a heat sink, wherein a so-called thermal base in the form of a vapor
chamber component using Vapor Chamber Technology ("VC-Tech") is provided in between
the heat pump and the heat sink in thermal contact therewith. Such thermal base provides
an improved spreading of heat across its entire cross section area in order to provide
a high thermal conductivity between the heat pump and the heat sink. However, even
though the transfer of heat between the heat pump and the heat sink can be improved
by the provision of such thermal base, the heat dissipation capabilities of the device
are limited by the heat transfer properties of the heat sink. As already mentioned
above, for this purpose, fans can be provided in order to improve the heat transfer
abilities of the heat sink. However, , heat sink designs which are usually cooled
by air with a fan are not only noisy, large and heavy but also mandatorily determine
a direction of the cooling air flow in close proximity to the mount and, thus, in
close proximity to the sample, which is not desired and, thus, require counter measures
to direct the air flow further away from the sample.
[0009] Also, the mounts of thermocycler are usually made of a solid material which requires
a smart overall thermal design in order to achieve a superior thermal homogeneity
of the mount due to the fact that already small temperature differences on the cooling
side of the mount result in thermal inhomogeneity on the sample receiving side of
the mount. In general, current thermocyclers use mostly forced air, extruded heat
sinks and sometimes also heat spreaders between the heat sink and the bottom side
of the heat pump, for example in the form of thermal interface plates, thermal interface
grease, or the above mentioned vapor chamber technology, wherein such heat sinks can
only provide limited heat transfer from the heat sink's fin surfaces into the cooling
air, wherein a maximizing of heat transfer rates generally results in large scale
heat sinks and, thus, in an increased size and weight of the thermocycler. Furthermore,
such a heat exchange setup is rather stiff and inflexible, since such large-sized
heat sink needs to be arranged directly underneath the heat pump and in almost direct
connection with its bottom side. In further detail, since mount heating/cooling is
based on thermal conductance, a homogeneous and/or at least symmetric placement of
thermal heating and cooling elements, such as the generally used TECs, is usually
required when implementing the solutions of the prior art. Due to these and other
problems and disadvantages, the prior art suggestions as presented above can not fulfill
the needs of users nowadays and, thus, do not provide satisfying solutions. Therefore,
the general need exists these days in the present technical field to provide a device
for simultaneous thermocycling biological samples with a further improved heating/cooling
performance in order to make diagnostic assays a lot faster and, thus, cheaper to
perform while maintaining or even improving precision as well as efficiency.
SUMMARY OF THE INVENTION
[0010] The present invention addresses the above described problems of improving the thermocycling
of biological samples by means of improved heating/cooling performance of a mount
of a thermocycling device. According to a first aspect of the present invention, a
device for thermocycling biological samples is provided, which device can be a thermocycler
or thermocycler unit for the simultaneous thermocycling of multiple samples to perform
multiple nucleic acid amplification reactions, wherein such device can be an internal
or integral part of a laboratory instrument, such as an analytical instrument or the
like, and more particularly wherein such device can be provided inside a housing of
such instrument.
[0011] The inventive device comprises a mount for receiving the biological samples, a heat
pump for heating and cooling the mount, the heat pump being thermally coupled to the
mount, a heat sink, and a control unit for controlling the thermocycling of the biological
samples. In general, the combination of mount, heat pump and heat sink can also be
referred to as thermal block unit of the inventive device. The heat sink of the device
comprises a primary heat exchanger with an inner space perfused by a heat exchanging
fluid, and a secondary heat exchanger, wherein the primary heat exchanger is thermally
coupled to the heat pump, and wherein the secondary heat exchanger is thermally coupled
to the heat pump through the primary heat exchanger. Here, the heat exchanging fluid
can be a heat exchanging liquid, such as a water-based cooling liquid medium, for
example a mixture of water and ethanol, wherein the inner space of the primary heat
exchanger provides a cavity where the liquid cooling medium can flow through for heat
transfer to the secondary heat exchanger. Accordingly, contrary to the commonly known
heat sinks, the heat sink of the present invention, which is used for heat dissipation
of the heat pump, is not implemented in the form of a commonly known heat sink with
fins, but is implemented in the form of a combination of primary heat exchanger provided
at the heat pump, i.e. the bottom side of the heat pump, and a secondary heat exchanger
which is provided at a different location, for example spaced apart from the heat
pump as well as from the primary heat exchanger, but is thermally coupled to the heat
pump via the primary heat exchanger. Thereby, by means of the heat exchanging fluid,
heat can be transported away from the primary heat exchanger and can be transferred
into the secondary heat exchanger for dissipation to ambient, or to laboratory infrastructure.
Thus, it becomes possible to rapidly transport heat away from the primary heat exchanger.
By the described heat exchanging fluid flow, the mount tempering can be operated at
optimum conditions, e.g. by bringing well cooled fluid into the primary heat exchanger
during cooling of the mount, by bringing warm fluid into the primary heat exchanger
during heating cycles of the mount, and/or by bringing adequate tempered fluid into
the primary heat exchanger during plateau phases to reduce any temperature differences
and, therefore, reduce thermal losses. Moreover, since the secondary heat exchanger
can be arranged well away from the PCR application site and in various orientations,
the secondary heat exchanger can be arranged in a flexible manner in numerous locations
within, at or outside of a thermal cycler, as desired.
[0012] In accordance with the present invention, the mount for receiving the biological
samples can be implemented as a mount comprising at least one flat surface or, alternatively,
a surface geometry with a plurality of wells, i.e. with a structured upper side comprising
recesses in order to provide a certain well geometry for receiving reaction vessels,
i.e. for receiving either one or a plurality of single reaction vessels or a plurality
of reaction vessels combined into a multi-well plate or the like. Also, the surface
of the mount can have a curved shape, and is preferably suitable for use with thermal
interface material. In case of the use of a multi-well plate, the same can be received
in the well geometry of the mount, wherein the mount as well as the respective multi-well
plate can provide, for example, 6, 12, 24, 48, 96, 384 or 1536 sample wells, or even
more. This means that the biological samples can be received in an array of reaction
vessels of a multi-well plate mounted in a respective recess structure of the mount
or on top of the mount. Alternatively, the mount can be a mount with a flat upper
side, i.e. the mount can substantially be a plate for receiving, on its surface, a
cartridge, a slide or any other kind of consumable comprising the biological samples,
or a microfluidic device comprising one or more flow channels connected to an array
of sample wells for receiving the biological samples. Moreover, the mount can comprise
either a flat lower side, or alternatively a structured lower side, wherein the lower
side of the mount can be integrated with a heat exchanger or with an additional heating/cooling
element. Furthermore, the mount of the present invention can use Vapor Chamber Technology
("VC-Tech"), for example in the form of a compact liquid heat exchanger provided within
the mount, such as a vapor chamber, i.e. a heat pipe for heat spreading and isothermalizing
in order to be able to absorb or provide required thermal energy, wherein a vapor
chamber is generally able to transport heat from a heat source to a heat sink with
a very small temperature gradient. Accordingly, the use of VC-Tech in the mount of
the present invention can achieve superior thermal homogeneity, even in case of undesired
uneven thermal heating and/or cooling. Also, in case of using fins within the vapor
chamber, a more effective heat transfer from liquid to the solid can be achieved,
and the high thermal capacity of water based cooling medium fins allows for a smaller
heat transfer surface and, therefore, miniaturized vapor chamber design.
[0013] With the implementation of vapor chamber technology inside the mount, the mount is
able to distribute any applied temperature much more evenly and rapidly to or from
the reaction vessels, i.e. the samples undergoing PCR. Thus, a superior vessel-to-vessel
uniformity can be provided, which can ensure consistent results over all reaction
vessels, wherein a fast transient response of all vessels on the same temperature
level right after reaching the target plateau temperature allows a reduction of the
plateau time and speeding up the whole PCR process.
[0014] Furthermore, in accordance with the present invention, a heat pump is generally to
be understood as a component that moves heat from one location, i.e. the heat source,
to another location, usually in the form of a sink or heat sink. Thus a heat pump
may be thought of as a "heater" if the objective is to warm the mount, or as a "cooler"
if the objective is to cool the mount. Usually, thermoelectric heating or cooling
by means of the Peltier effect is used to create a heat flux. As an example, a thermoelectric
cooler "TEC", preferably in the form of a Peltier element, is used as a solid-state
active heat pump with consumption of electrical energy, depending on the direction
of the current, which heat pump transfers heat from its one side to the other, for
example in order to provide heat to the mount during heating phases, such as during
PCR denaturation, or to retract heat from the mount during cooling phases, such as
during PCR annealing, and provide the heat to a sink or heat sink.
[0015] A heat sink in the sense of the present invention is to be understood as a passive
heat exchanging structure that transfers heat or cold generated by an electronic or
a mechanical component, such as the above described heat pump, to a fluid medium,
often air or a liquid coolant, where it is dissipated away from the heat pump, thereby
allowing regulation of the heat pump's temperature at optimal levels. Heat sinks are
often used with components where the heat dissipation ability of the component itself
is insufficient to moderate its temperature, or at least insufficient to moderate
its temperature within the desired time frame, as is often the case with heat pumps
in PCR applications. A heat sink in the classical sense is designed to maximize its
surface area in contact with the cooling medium surrounding it, such as air. Air velocity,
choice of material, protrusion design and surface treatment are factors that affect
the performance of a heat sink. The heat sink in accordance with the present invention,
however, comprises a plurality of heat exchangers, i.e. at least a proximate or primary
heat exchanger and a remote or secondary heat exchanger, with both heat exchangers
being able to transfer heat or cold to a fluid medium, such as a heat exchanging fluid,
for example in the form of a liquid coolant, to dissipate the heat or cold away from
the source, i.e. the heat pump. Here, the heat exchanging fluid perfuses an inside
or inner space of the primary heat exchanger, wherein the "inner space" defines a
space internal to the primary heat exchanger in or through which the heat exchanging
fluid can flow/stream. Accordingly, the primary heat exchanger is based on a fluid
heating and/or cooling system, such as a liquid cooling system, wherein the fluid
heating and/or cooling system enables a low noise design of the heat sink, which is
light in weight and comprises a compact design, but still allows for efficient heating/cooling.
Thus, the thermal capacitance of the used fluid and/or the design of the primary heat
exchanger achieve an optimized heat transfer to and from the heat pump and, thus,
to and from the mount.
[0016] According to a specific embodiment of the present invention, the mount of the device
for thermocycling biological samples is connected to the heat pump and/or the heat
pump is connected to the primary heat exchanger, for example by means of a releasable
force-fit connection, such as clamping. Here, according to a further specific embodiment,
the mount, the heat pump and the primary heat exchanger are clamped together in order
to form a tight connection therebetween, thereby constituting a thermal block unit
as already mentioned above. By means of the clamped combination of components, homogenous
contact pressure can be achieved between these components, whereby homogenous thermal
contact can be established between the mount, the heat pump and the primary heat exchanger,
i.e. between the lower side of the mount and the upper side of the heat pump as well
as between the lower side of the heat pump and the upper side of the primary heat
exchanger. For the clamping of the mentioned components, a clamping mechanism can
be provided as part of the inventive device, which can be implemented in the form
of a threaded connection, wherein, for example, threaded bolts or screws are guided
through through-holes provided inside the primary heat exchanger and through-holes
or gaps within the heat pump and are screwed into respective counterparts, such as
threaded holes provided in the mount, wherein the screwing connection between bolts/screws
and mount can be further fixed by means of adhesive or the like. Additionally, on
the side of the primary heat exchanger facing away from the mount, the bolts/screws
can be further provided with disc springs or the like, fixed by means of screw heads
or additional screw nuts, in order to provide a distinctive clamping pressure on the
thus connected components of the inventive device by means of a distinct torque to
be applied onto the screw nuts.
[0017] According to a further specific embodiment of the present invention, the secondary
heat exchanger is thermally connected to the primary heat exchanger by means of a
heat exchanging fluid circulation system for circulating the heat exchanging fluid
between the primary heat exchanger and the secondary heat exchanger. Thereby, it becomes
possible to stream hot or cold heat exchanging fluid away from the primary heat exchanger,
heat or cool the same by means of the secondary heat exchanger, and recirculate the
heated or cooled heat exchanging fluid back to the primary heat exchanger. Thus, it
becomes possible to adjust the temperature level of the heat exchanging fluid inside
the inner space of the primary heat exchanger by means of a component provided or
arranged away from the heat source, i.e. the heat pump, and, thus, to react fast to
undesired temperature levels at the primary heat exchanger. As an alternative to the
connection between the primary heat exchanger and the secondary heat exchanger by
means of the above described heat exchanging fluid circulation system, the secondary
heat exchanger can alternatively or additionally be thermally connected to the primary
heat exchanger by means of a heat-transfer component such as a heat pipe or the like,
which can further improve the function of the heat sink in regard to a fast control
of the temperature of the heat pump. Here, the inner space or cavity of the primary
heat exchanger can provide a location where the heat is collected and transferred
to the heat pipe, to be transported to the secondary heat exchanger. In line with
a further preferred embodiment of the present invention, the circulation system can
comprise a tubing connection between the primary heat exchanger and the secondary
heat exchanger for circulatory transfer of the heat exchanging fluid between the primary
heat exchanger and the secondary heat exchanger. Here, the tubing connection is established
by means of one or several tubes, supporting the arrangement of the secondary heat
exchanger spaced apart from the primary heat exchanger, wherein the tubing connection
can be implemented in the form of a circular tubing connection in order to be able
to discharge the heat exchanging fluid out of the inner space of the primary heat
exchanger and towards the secondary heat exchanger, pass the heat exchanging fluid
along the secondary heat exchanger, or also into an inner space provided inside the
secondary heat exchanger, and recirculate the same to and into the primary heat exchanger.
[0018] Accordingly, the heat exchanging fluid, for example a liquid cooling medium, can
transport heat away from the primary heat exchanger and transfers it into the secondary
heat exchanger for dissipation of the heat to ambient conditions at the location of
the secondary heat exchanger which can be arranged outside the thermocycler, or for
dissipation of the heat to adjacent laboratory infrastructure, for example a ventilation
system, a cooling system or the like.
[0019] Now, the fluid volume already provided inside the tubing system may already be regarded
as an initial fluid reservoir. However, and in accordance with a further specified
embodiment of the circulation system of the present invention, at least one (additional)
fluid reservoir accommodating heat exchanging fluid of a predetermined temperature
can be provided as part of the circulation system, i.e. can be fluid-connected with
the circulation system, or better with the tubing connection, which can be used, on
demand, in case heat exchanging fluid of a certain temperature as stored inside the
fluid reservoir can be used to heat or cool the heat exchanging fluid streaming inside
the circulation system. Here, the circulation system can comprise a plurality of fluid
reservoirs accommodating fluid of different predetermined temperatures. Accordingly,
any such reservoir can be built as an extra volume in the liquid path provided by
the circulation system. For example, one fluid reservoir can store heated heat exchanging
fluid, and another fluid reservoir can store cooled heat exchanging fluid, and each
fluid reservoir can add stored heat exchanging fluid into the circulation system on
demand. According to a specific embodiment of the mentioned tubing connection, the
used tubes can be made of an inner layer of ethylene propylene diene monomer (EPDM)
rubber and an outer layer of nitrile butadiene (NBR) rubber, preferably enforced with
a synthetic mesh, or can be generally made of EPDM, NBR, fluorinated ethylenepropylene
polymer (FEP), Polytetrafluoroethylene (PTFE), Polyvinyl chloride (PVC), polyethersulfone
(PES), fluoroelastomer (FKM), silicone, or can also generally made of metal tubes
or the like, or can additionally be jacketed by means of a heat isolating material.
[0020] Moreover, according to a further specific embodiment of the tubing connection, the
tubing connection can be established by flexible tubing, i.e. one or several flexible
tubes or flexible pipes, in order for the secondary heat exchanger to be variably
positioned in relation to the primary heat exchanger, for example far away from the
primary heat exchanger and, thus, far away from the heat pump. Accordingly, since
the secondary heat exchanger can be provided within a housing of the instrument accommodating
the thermocycler, such as an analytical instrument, and can be placed therein independently
of the primary heat exchanger, it becomes possible to position the secondary heat
exchanger, for example, near a front part of the inner space of the housing. Such
positioning makes it possible to unobstructedly suck cool ambient air, for example
by means of a fan or the like, via a ventilation opening, a ventilation grille or
the like, which is provided on the front side of the housing. Furthermore, the described
variable positionability of the secondary heat exchanger independently from the primary
heat exchanger results in the fact that the risk of warm air generated by neighboring
laboratory appliances being sucked into the housing can be avoided due to the possibility
of arranging the secondary heat exchanger in a location away from the warm air source,
and the cooling efficiency of the thermal mount unit can be significantly improved.
As a particular example, the presently described improved cooling concept based on
the possibility of a forced airflow input from the front side of the housing can be
implemented by means of a forced input of air entering through a ventilation opening
provided at the top and/or bottom of a loading flap of the housing, and is further
supported by a hot air outlet, e.g. another ventilation opening, provided at the back
side of the thermocycler in a respective suitable position. Accordingly, (cool) ambient
air can be introduced through a top and/or a bottom of the loading flap at a front
side of the housing, and a hot air outlet provided at the back side of the housing
can discharge the hot air to be removed from the thermocycler unit. Thus, any kind
of cooling fan provided for the heat sink can now be integrated directly with the
thermocycler unit, and does not necessarily have to be provided integrally with the
housing or the loading flap anymore, resulting in a fan integration within the housing's
inner space, and directly at the thermocycler unit, achieving a significantly reduced
noise level for the benefit of a user as well as a design advantage since the ventilation
openings, i.e. the air supply, can be hidden anywhere in the housing.
[0021] Basically, the presently described heating/cooling concept of a thermocycler unit
within the housing of an analytical instrument allows a free arrangement of the second
heat exchanger as well as its respective fan, if any, around the thermocycler unit
inside the housing. For example, the fan can sit directly at or on the secondary heat
exchanger, making it an integral part of the thermocycler unit and, thus, resulting
in the fact that the entire thermocycler unit is a secluded and, thus, interchangeable
unit within the analytical instrument. Moreover, due to the fact that the presently
described solution achieves sufficient heating/cooling performance in order to omit
the provision of any kind of extruded heat sink element arranged directly underneath
the TECs, sufficient air flow areas exists inside the housing, so that the fan does
not have to arranged directly at the ventilation openings but can be arranged in a
distance away from any ventilation opening. This is advantageous since such further
distance from the housing, preferably supported by a certain deflection of air flow
within the housing, helps to further reduce the mentioned noise level.
[0022] According to a further specific embodiment of the present invention, at least one
fluid pump, such as a liquid pump, is connected to the primary heat exchanger and
the secondary heat exchanger for pumping the heat exchanging fluid and controlling
the flow speed of heat exchanging fluid in the heat sink, i.e. in the previously described
circulation system between the primary heat exchanger and the secondary heat exchanger.
The fluid pump of the device can be used to achieve fluid flow, and can be controllable,
e.g. the pump performance as well as the direction of flow can be controlled. Furthermore,
the fluid pump can be arranged between the primary heat exchanger and the secondary
heat exchanger, and can be connected within the circulation system, such as a connecting
part within the tubing connection as described above, in order to be providing the
function of pumping heat exchanging fluid from the primary heat exchanger to the secondary
heat exchanger, and/or vice versa. By the regulation of the heat exchanging fluid
pump, a heating or cooling capacity within the primary heat exchanger can be well
controlled, wherein the addition of different temperature level fluid reservoirs as
described above can further improve the controllability of the heating or cooling
capacity of the heat sink, since the contents of such fluid reservoirs are very fast
available and can be transported by means of the fluid pump into the primary heat
exchanger.
[0023] According to a further specific embodiment of the present invention, a thermal coupling
between the mount and the heat pump and/or between the heat pump and the primary heat
exchanger is established by means of a thermal interface material, wherein any such
thermal interface material can comprise at least one of a carbon based material, such
as graphite, for example pure pyrolytic graphite, or graphene, a silicone compound,
a phase change material and thermal grease, or a combination thereof. In general,
any thermal coupling connection between components of the presently described device
can be provided with such thermal interface material, in order to ensure a sufficient
thermal conductivity at the contact surfaces of such components. Accordingly, any
heating or cooling component is thermally connected to its adjacent component or components
by means of using thermal interface material as mentioned before, in order to enhance
the thermal coupling between any of those coupled components.
[0024] Furthermore, in accordance with the present invention, the primary heat exchanger
can be a so called solid-to-liquid heat exchanger, i.e. a heat exchanger in which
heat is transferred between a solid object, such as the outer hull of the primary
heat exchanger comprising an inner space, and a liquid such as one example of the
heat exchanging fluid provided inside the inner space of the primary heat exchanger,
wherein the primary heat exchanger in the form of a solid-to-liquid heat exchanger
can be a compact liquid heat exchanger providing a compact design. In this regard,
the inner space of the primary heat exchanger can comprise one or more projections
for surface enlargement, such as fins, provided on the inner surface of the inner
space of the primary heat exchanger and surrounded by the heat exchanging fluid, wherein
the fins can be pin fins or swage fins, or any other suitable kind of respective surface
enlarging projection. Here, the secondary heat exchanger can comprise a similar structure
as the primary heat exchanger. Additionally, the secondary heat exchanger may also
be provided with external fins, such as known from usual heat sinks. Moreover, the
primary heat exchanger can consist of one or more parts made from copper or other
suitable thermally conducting materials, such as aluminum or silver, wherein the solid
parts of the primary heat exchanger constitute the parts made from thermally conducting
material. Here, the part of the primary heat exchanger which is not directly connected
to any heat producing components and/or the mount can consist of a thermally conducting
material, but does not have to consist of such a thermally conducting material, which
can improve the design and weight of the primary heat exchanger.
[0025] Moreover, the primary heat exchanger can be designed with different layers of material,
and the layers can be made of different material, for example one material for an
upper layer or layers, and another material for lower layers. Also, the primary heat
exchanger's side without contact to the mount or the heat pump can be made out of
a plastic or polymer material without any thermal relevance. Also, a thickness of
an interface side of the primary heat exchanger can be used as a thermal reservoir.
Additionally or alternatively, the primary heat exchanger can also comprise at least
one through hole provided for allowing an assembly of the device in the manner of
a thermally sandwiched structure, meaning that all components of the inventive device
can be assembled to form a thermally sandwiched structure by means of respective through
holes and respective connectors, such as screws or the like, in order to force the
components of the device to be sandwiched together.
[0026] According to a further specific embodiment of the present invention, with particular
view on the secondary heat exchanger, the same can comprise a temperature sensor for
controlling its heat exchange rate, wherein the detection values of the temperature
sensor can be used by a control unit or the like to counteract any undesired measured
heat exchange rate values. Furthermore, the secondary heat exchanger can comprise
an inner space containing heat exchanging fluid pumped through its inner space, similar
to the primary heat exchanger, wherein a heat exchange rate of the secondary heat
exchanger can be controlled by means of a pump speed sensor which controls the speed
of the fluid pumped through the inner space of the secondary heat exchanger, and/or
a volume flow sensor, also referred to as fluid flow sensor, which controls the flow
volume flowing through the secondary heat exchanger. In accordance with the necessity
to change the heat exchange rate of the secondary heat exchanger based on the measured
pump speed sensor values and/or volume flow sensor values, the fluid pump pumping
the heat exchanging fluid through the secondary heat exchanger can be controlled to
pump more or less fluid therethrough, in order to change the heat exchange rate to
the desired level controlled by the pump speed sensor and/or the volume flow sensor.
Moreover, the secondary heat exchanger can also interact with a working fluid for
cooling the secondary heat exchanger, wherein the heat exchange rate of the secondary
heat exchanger is controlled by means of a working fluid pump, meaning that the secondary
heat exchanger can be connected to a working fluid by means of an interior channel,
a working fluid pipe or the like, wherein the secondary heat exchanger can be heated
and/or cooled not only indirectly by the heat exchanging fluid received from the primary
heat exchanger or the fluid reservoirs within the circulation system, but can also
be actively heated or cooled by means of the working fluid, which can be a cooling
fluid or the like.
[0027] Alternatively or additionally, the secondary heat exchanger can interact with a fan
and/or a radiator, in particular in case the secondary heat exchanger is provided
with external fins or the like in order to enlarge the overall surface area of the
secondary heat exchanger, wherein the heat exchange rate of the secondary heat exchanger
is controlled by means of a fan speed sensor and/or a radiator temperature sensor.
Alternatively or additionally, another kind of fluid cooling structure or mechanism
can also be provided for the secondary heat exchanger, or also another element with
a different temperature in thermal connection with the secondary heat exchanger, for
dissipation of heat. Accordingly, in case the secondary heat exchanger dissipates
heat to ambient air by e.g. a radiator or a fan, or to laboratory infrastructure,
such as a cooled water circuit or a different cold/heat reservoir, its heat exchange
rate is monitored and can be affected by means of a respective control based on the
monitoring results, such as provided directly from the secondary heat exchanger temperature
sensor, or indirectly from the pump speed sensor, the volume flow sensor, the fan
speed sensor and/or the radiator temperature sensor. In general, all components of
the inventive device can be equipped with sensors, such as temperature sensors, flow
sensors, vibration sensors, pressure sensors, etc.. Also, the ambient air temperature
can be monitored, for example by means of an ambient sensor monitoring, for example
humidity and/or ambient temperature, wherein a temperature difference between ambient
temperature and the temperature of the heat exchanging fluid can be monitored, in
order to control heat transfer in an accurate manner.
[0028] According to a further specific embodiment of the present invention, with particular
view on the mount of the inventive device, the mount can be made of solid material,
such as silver or aluminum, solid ceramic, a carbon based material or any combination
thereof, and can function as a heat conductor for achieving thermal homogeneity, as
also already described further above. Alternatively, or also in part additionally,
provided that the mount can consist of several parts and materials, the mount can
be made of solid thermal conductive material, such as copper, and can comprise a vapor
chamber (VCM; 3D heat pipe) for transporting and distributing heat between the heat
pump and the biological samples in a uniform manner. Here, in case the mount provides
for a vapor chamber, then the heat pump can be placed at almost any location of the
mount, not necessarily between mount and primary heat exchanger, due to the excellent
thermal conductivity of a vapor chamber. One example of such a vapor chamber mount
is described in
WO 2013/075839 A2.
[0029] According to a further specific embodiment of the present invention, with particular
view on the heat pump, the same can be one of a thermoelectric device, a resistive
heater, or a combination thereof, wherein the thermoelectric device can be a Peltier
element. Alternatively or additionally, the resistive heater can be one of a ceramic
heater, a heating foil, a heating cartridge or a wire wound heater. Moreover, the
heat pump can be arranged between the mount and the primary heat exchanger, for example
in a sandwiched manner, as already indicated above in relation to another specific
embodiment of the present invention. Furthermore, the heat pump can be arranged at
least in part inside a recess provided in the mount, in more detail in a recess provided
in the side of the mount opposite of the side receiving the biological samples.
[0030] In general, a perimeter heater or edge heater can also be provided at the mount in
order to minimize any thermal non-uniformity across the samples or any kind of sample
holder. Such perimeter heater can be positioned at the side of or on top of the mount
and around a sample holder, such as a multi-well plate, to counter any heat loss at
its edges. Additionally, a heated cover can be provided, designed to keep the reaction
vessels closed during thermal cycling and to heat the upper portion of such vessels
to prevent condensation and the like.
[0031] According to a further specific embodiment of the present invention, the inventive
device can further comprise at least one sensor for controlling the temperature at
the biological samples received in the mount, wherein such sensor can be a temperature
sensor, a fan speed sensor or a fluid flow sensor, and wherein such sensor can be
provided at the mount, at the heat pump, at the primary heat exchanger, and/or at
the secondary heat exchanger. Thus, by providing a respective sensor at any of the
components of the inventive device, the temperature of the biological samples during
PCR can be monitored closely, and the heating/cooling function of the device can be
controlled based on the measured temperature values of the samples, in order to regulate
the different temperature plateaus of the PCR accurately and efficiently. Accordingly,
such sensor(s), for example temperature sensors, enable a precise control of the mount
temperature by a control algorithm or the like, wherein sensors in the heat exchangers,
such as fan speed sensors, temperature sensors, and other sensors are used to control
their respective heating/cooling rate, wherein the heating/cooling power can be significantly
varied by changing fan speed and/or fluid pump speed. For example, a heat sink temperature
measurement can be carried out by the mentioned control algorithm, for linearizing
thermal output power from the heat pump.
[0032] In other words, when summarizing the above, a device for PCR application for one
or more samples is suggested, wherein the samples are placed on a mount with or without
well geometry, wherein the device includes the mount, one or more heater elements
at any position of the mount, a primary heat exchanger, a secondary heat exchanger
for exchanging heat to ambient air or laboratory infrastructure, and a connecting
element between the primary and the secondary heat exchangers, for example in the
form of cooling liquid tubes or the like, or, alternatively or additionally, a heat
pipe, wherein the thermal layers of the device, i.e. its components, can be thermally
connected via thermal interface material, and wherein the thermal components can be
equipped with sensors, such as temperature sensors, fan speed sensors, fluid flow
sensors, volume flow sensors or the like, to control the temperature at the samples
in or on the mount.
[0033] According to another aspect of the present invention, an instrument for simultaneously
monitoring multiple nucleic acid amplification reactions during thermocycling biological
samples is provided, wherein the instrument comprises a device for thermocycling the
biological samples as described previously, and wherein the instrument further comprises
one or several excitation light sources for applying excitation energy to the nucleic
acid amplification reactions occurring inside one or several reaction vessels received
by the mount, and one or more sensors for simultaneously detecting light emitted from
the multiple nucleic acid amplifications. Such an instrument can be structured similar
to the already known Roche LightCycler® instruments which provide high-performance,
and high-throughput PCR platforms which can be used, for example, for gene detection,
gene expression analysis, genetic variation analysis, and array data validation. Such
benchtop instrument solutions comprise, inter alia, a plate-based real-time PCR device
as a robotically controllable, automated high-throughput solution for general laboratory
use.
[0034] Usually, an automated processing system, such as an analytical, pre-analytical or
post-analytical processing system, which is commonly employed in state-of-the-art
laboratories for automatically processing biological sample, can comprise the previously
described device, or even the entire previously described instrument. Here, the term
"laboratory instrument" or "instrument" of the laboratory encompasses any apparatus
or apparatus component operable to execute one or more processing steps / workflow
steps on one or more biological samples, and covers analytical instruments, pre-analytical
instruments, and also post-analytical instruments. The expression "processing steps"
thereby refers to physically executed processing steps, such as conducting the particular
steps of a PCR conduct. The term "analytical" as used herein encompasses any process
step carried out by one or more laboratory devices or operative units which are operable
to execute an analytical test on one or more biological samples. In the context of
biomedical research, analytical processing is a technical procedure to characterize
the parameters of a biological sample or of an analyte. Such characterization of parameter
comprises, for example, the determination of the concentration of particular proteins,
nucleic acids, metabolites, ions or molecules of various sizes in biological samples
derived from humans or laboratory animals, or the like. The gathered information can
be used to evaluate e.g. the impact of the administration of drugs on the organism
or on particular tissues. Further analyses may determine optical, electrochemical
or other parameters of the samples or the analytes comprised in a sample.
[0035] According to another aspect of the present invention, a method for thermocycling
biological samples using a device for thermocycling the biological samples as described
previously is provided with the present invention, wherein the inventive method comprises,
inter alia, the step of monitoring the temperature at the biological samples, for
example by means of the sensors as described further above, and the step of controlling
the output of the heat pump and/or the heat exchange rate of the primary heat exchanger
and/or the output of any further heater, such as an edge heater, wherein the controlling
step is carried out based on the detected and monitored temperature at the biological
samples resulting from the monitoring step. With such method, a thermal performance
of the inventive device can be actively influenced by controlling the heat pump or
any additional heaters, by controlling the performance of the heat sink, i.e. the
primary heat exchanger and/or the secondary heat exchanger, and/or by monitoring and
exploiting further environmental parameters or design parameters as mentioned above.
[0036] According to a specific embodiment of the above described method, the above described
step of controlling the heat exchange rate of the primary heat exchanger can be executed
by the control unit of the described device, wherein the controlling step can comprise
a control of the heat exchange rate of the second heat exchanger and/or a stop or
reverse of a flow of heat exchanging fluid through the inner space of the primary
heat exchanger. Moreover, according to a further specific embodiment of the present
invention, the step of controlling the heat exchange rate of the primary heat exchanger
can also include the control of fluid speed to cool the heat exchange fluid in the
heat sink above ambient temperature and approximate to the predetermined lowest temperature
of the thermocycles controlled by the control unit. Accordingly, a thermocycler specific
method feature for the controlling step is, for example, the stop of a flow of heating
or cooling liquid in the heat sink at certain PCR protocol steps, which can minimize
thermal losses during the PCT temperature plateau phases. Moreover, the active control
of the temperature of the heat exchanging fluid, such as cooling liquid flowing through
the heat sink, can be used to control the overall thermal performance of the device.
As a further example, reversing the fluid flow can be used to fill the primary heat
exchanger and/or the secondary heat exchanger with a heat exchanging fluid with either
a higher or lower temperature compared to the fluid currently in place inside the
primary heat exchanger and/or the secondary heat exchanger. Additionally or alternatively,
the inside of the already described fluid reservoirs can be refilled or replenished
with fluid of a certain (high or low) temperature. Also, additionally or alternatively,
the flow path as defined by the circulation system, i.e. the tubing system, optionally
together with the fluid reservoirs and pumps, if any, can be modified in a certain
way, for example by inclusion of additional tubing, reservoirs and pumps, in order
to achieve a certain control effect, for example by prolonging the flow path, increasing
the flow speed, and/or enlarging the amount of fluid provided inside the flow path
of the heat exchanging fluid.
[0037] The above described method steps can be controlled by the control unit of the described
device, which can also control any kind of actuation or monitoring of the above described
device and its components, wherein the term "control unit" as used herein encompasses
any physical or virtual processing device, such as a CPU or the like, which can also
control the entire instrument or even an entire workstation comprising one or more
laboratory instruments in a way that workflow(s) and workflow step(s) are conducted.
The control unit may, for example, carry different kinds of application software and
instruct the automated processing system or a specific instrument or device thereof
to conduct pre-analytical, post analytical and analytical workflow(s)/ workflow step(s).
The control unit may receive information from a data management unit regarding which
steps need to be performed with a certain sample. Further, the control unit might
be integral with a data management unit, may be comprised by a server computer and/or
be part of one instrument or even distributed across multiple instruments of the automated
processing system. The control unit may, for instance, be embodied as a programmable
logic controller running a computer-readable program provided with instructions to
perform operations. Here, in order to receive such instructions by a user, a user
interface can additionally be provided, wherein the term "user interface" as used
herein encompasses any suitable piece of application software and/or hardware for
interactions between an operator and a machine, including but not limited to a graphical
user interface for receiving as input a command from an operator and also to provide
feedback and convey information thereto. Also, a system / device may expose several
user interfaces to serve different kinds of users / operators.
[0038] As used herein and also in the appended claims, the singular forms "a", "an", and
"the" include plural reference unless the context clearly dictates otherwise. Similarly,
the words "comprise", "contain" and "encompass" are to be interpreted inclusively
rather than exclusively; that is to say, in the sense of "including, but not limited
to". Similarly, the word "or" is intended to include "and" unless the context clearly
indicates otherwise. The terms "plurality", "multiple" or "multitude" refer to two
or more, i.e. 2 or >2, with integer multiples, wherein the terms "single" or "sole"
refer to one, i.e. =1. Furthermore, the term "at least one" is to be understood as
one or more, i.e. 1 or >1, also with integer multiples. Accordingly, words using the
singular or plural number also include the plural and singular number, respectively.
Additionally, the words "herein," "above,", "previously" and "below" and words of
similar import, when used in this application, shall refer to this application as
a whole and not to any particular portions of the application.
[0039] The description of specific embodiments of the disclosure is not intended to be exhaustive
or to limit the disclosure to the precise form disclosed. While the specific embodiments
of, and examples for, the disclosure are described herein for illustrative purposes,
various equivalent modifications are possible within the scope of the disclosure,
as those skilled in the relevant art will recognize. Specific elements of any foregoing
embodiments can be combined or substituted for elements in other embodiments. Also,
in drawings, same reference numerals denote same elements to avoid repetition, and
parts readily implemented by one of ordinary skill in the art may be omitted. Furthermore,
while advantages associated with certain embodiments of the disclosure have been described
in the context of these embodiments, other embodiments may also exhibit such advantages,
and not all embodiments need necessarily exhibit such advantages to fall within the
scope of the disclosure as defined by the appended claims.
[0040] The following examples are intended to illustrate various specific embodiments of
the present invention. As such, the specific modifications as discussed hereinafter
are not to be construed as limitations on the scope of the present invention. It will
be apparent to the person skilled in the art that various equivalents, changes, and
modifications may be made without departing from the scope of the present invention,
and it is thus to be understood that such equivalent embodiments are to be included
herein. Further aspects and advantages of the present invention will become apparent
from the following description of particular embodiments illustrated in the figures.
BRIEF DESCRIPTION OF THE DRAWINGS
[0041]
- Figure 1
- is a conceptual illustration of a device for thermocycling biological samples according
to an embodiment of the present invention;
- Figure 2
- is a schematic structural illustration of the device for thermocycling biological
samples according to the embodiment as shown in fig. 1;
- Figure 3
- is a schematic functional illustration of a device for thermocycling biological samples
according to another embodiment of the present invention;
- Figure 4
- is a schematic functional illustration of a modified device for thermocycling biological
samples according to the embodiment as shown in fig. 3; and
- Figure 5
- is a flowchart of a method using the device according to an embodiment of the present
invention.
LIST OF REFERENCE NUMERALS
[0042]
- 1
- device for thermocycling biological samples
- 1'
- device for thermocycling biological samples
- 2
- mount / sample tempering mount
- 21
- mount's upper flat surface
- 2'
- mount / sample tempering mount
- 22'
- indentation / well
- 3
- heat pump / TEC / heater
- 3'
- heater
- 31
- thermal interface material TIM
- 4
- heat sink (including primary and secondary heat exchanger)
- 4'
- heat sink (including primary and secondary heat exchanger)
- 41
- heat exchanging fluid / heating/cooling liquid
- 5
- primary heat exchanger
- 5'
- primary heat exchanger
- 51
- primary heat exchanger's inner space
- 51'
- primary heat exchanger's inner space
- 511'
- fins
- 52'
- screw
- 53'
- through hole in primary heat exchanger
- 6
- secondary heat exchanger
- 6'
- secondary heat exchanger
- 61'
- fins
- 62'
- fan
- 7
- control unit
- 8
- heat exchanging fluid circulation system
- 81
- fluid reservoir
- 82
- fluid pump
- 9
- sensor
- 91
- temperature sensor
- 92
- fluid flow sensor
- 100
- monitoring step
- 200
- controlling step
- 300
- heat exchanging fluid flow controlling step
- 400
- fluid reservoir controlling step
DETAILED DESCRIPTION
[0043] Fig.1 shows a functional concept of a device 1 for thermocycling biological samples
according to an embodiment of the present invention, wherein the device 1 is intended
for PCR applications, for example as one main component of an analysis instrument.
As can be gathered from the schematic illustration of fig. 1, the device 1 comprises
a sample tempering mount 2 and a heat pump 3, which are illustrated in fig. 1 as one
functional unit for the sake of simplicity, wherein the mount 2 can also be implemented
as a mount including a heat pipe in the form of a vapor chamber or the like. Here,
the mount 2 can comprise a structured upper side with recesses in order to provide
a certain well geometry for receiving reaction vessels, or with a flat upper side
for receiving a cartridge, a slide or any other kind of consumable bearing the biological
samples to be thermocycled, or a microfluidic device comprising one or more flow channels
connected to an array of sample wells for receiving the biological samples. Moreover,
the device 1 comprises a heat sink 4 consisting, in a functional sense, of a primary
heat exchanger 5 and a secondary heat exchanger 6, wherein the mount 2, the heat pump
3 and the primary heat exchanger 5 constitute a functional unit which can be sandwiched
together, for example by means of a clamping mechanism for clamping these components
together in order to achieve a tight thermally-connected unit. Furthermore, the mount
2, the heat pump 3 and the primary heat exchanger 5 can comprise a thermal interface
material 5 thereinbetween, wherein, in use of the device 1, thermal interface material
31 is only depicted in fig. 1 between the lower side of the mount-heat pump-combination
2, 3, i.e. the lower side of the heat pump 3, and the upper side of the primary heat
exchanger 5, in order to improve the thermal conductivity between these components.
[0044] As can also be gathered from fig. 1, the primary heat exchanger 5 is connected to
the secondary heat exchanger by a heat exchanging fluid circulation system 8, wherein,
in fig. 1, only a single line stands for the circulation system 8 circulating heat
exchanging fluid between the two heat exchangers 5, 6, i.e. within the heat sink 4.
In particular, the secondary heat exchanger 6 can be arranged away from the primary
heat exchanger 5 without disrupting the functional unit of the heat sink 4, since
a thermal connection between the primary heat exchanger 5 and the secondary heat exchanger
6 is established by a connecting element, such as fluid-bearing tubes or a heat pipe,
constituting a circulation system 8 for heat exchanging fluid. Moreover, as depicted
in fig. 1, the device 1 comprises a particular monitoring system based on a plurality
of sensors 9, which are for control and prediction of the overall tempering of the
device 1. Here, a temperature sensor 91, or alternatively a heat flux sensor or the
like, is provided at the unit of mount 2 and heat pump 3 for measuring the temperature
of mount 2 and heat pump 3, another temperature sensor 91 is provided on the thermal
interface material 31 for measuring its temperature, a further temperature sensor
is provided at the primary heat exchanger 5 for measuring its temperature, a further
temperature sensor 91 is provided at the secondary heat exchanger 6 for measuring
its temperature, and a fluid flow sensor or volume flow sensor 92 is provided at the
heat exchanging fluid circulation system 8 for measuring the fluid flow, i.e. the
flow velocity of the heat exchanging fluid circulating inside the heat exchanging
fluid circulation system 8. Here, the sensor 92 might also be able to measure the
temperature of the heat exchanging fluid, or the pump speed of a pump (not shown)
provided in the fluid circulation system 8, in order to improve the monitoring capability
of the device 1. All sensors 9 are connected to a control element or control unit
7, for example in the form of a CPU 7, which is able to receive the measuring signals
from the sensors 9 and process the same, in order to monitor the temperatures at different
locations of the device 1, and to control and actively influence the temperature of
the samples at the mount 2 in regard to the PCR thermocycling requirements.
[0045] As a more concrete illustration, fig. 2 depicts the concept of the device 1 of fig.
1 in a structural manner. Accordingly, it can be gathered from fig. 2 that the device
1 comprises the mount 2 with a flat surface 21, wherein several heating/cooling elements,
depicted in form of heat pumps or TECs 3, such as Peltier elements, are provided in
between the mount 2 and the primary heat exchanger 5 in the form of a solid-to-liquid
heat exchanger. Reference sign 3 in fig. 2 can also illustrate the position of a heating
element, as an alternative to a Peltier element. Furthermore, additional heating/cooling
elements are depicted in fig. 2 on a side surface of the mount 2, or also on the flat
surface 21 of the mount 2, which elements 3' are preferably implemented as heaters,
such as multi-well plate edge heaters or the like. Thereby, a comprehensive heating/cooling
of the mount 2 can be achieved, wherein any thermal connection between the heat pumps
3 and the mount 2, as well as a thermal connection between the heaters 3' and the
mount 2 can be enforced by the use of thermal interface material 31. Also, a connection
surface between the heat pumps 3 arranged in between the mount 2 and the primary heat
exchanger 5 are provided with thermal interface material 31, in order to enhance any
temperature conductivity between the mount 2, the heat pumps 3 and the heat sink 4,
constituting a thermal block unit, wherein the components of the thermal block unit,
i.e. the mount 2, the primary heat exchanger 5 and the heat pumps 3 arranged therebetween
can be clamped together in order to establish a tight thermally conducting assembly.
A respective clamping mechanism is described further below in regard to figs. 3 and
4. From fig. 2, similar to the concept as shown in fig. 1, it can be clearly taken
that the remaining part of the heat sink 4, i.e. the secondary heat exchanger 6, can
be arranged in a spaced-away manner distant from the primary heat exchanger 5, wherein
the secondary heat exchanger 6 used to support a temperature compensation function
of the primary heat exchanger 5 in regard to the mount 2 can be arranged far away
from the thermal block unit in order to be thermally independent therefrom.
[0046] As can also be gathered from fig. 2, the primary heat exchanger 5 comprises an inner
space 51 filled with heat exchanging fluid 41, such as a cooling liquid or the like,
and the primary heat exchanger 5, or better the inner space 51 of the primary heat
exchanger 5, is in fluid connection with the secondary heat exchanger 6 by means of
a connecting element, such as a flexible tubing system or the like, constituting the
heat exchanging fluid circulation system 8. In order to be able to circulate the heat
exchanging fluid 41 from the primary heat exchanger 5 to the secondary heat exchanger
6, and back, a fluid pump 82 is provided as part of the heat exchanging fluid circulation
system 8, and is, thus, arranged in between the primary heat exchanger 5 and the secondary
heat exchanger 6. Accordingly, in case the temperature of the samples is too high,
or in case the sample temperature needs to be decreased in order to achieve a lower
temperature plateau during PCR cycling, the temperature of the mount 2 needs to be
decreased rapidly by means of the heat pumps 3, and the removed heat must be removed
from the heat pumps 3. In such case, the primary heat exchanger 5 acquires the heat
to be removed, transfers the same to the heat exchanging fluid 41. The heated-up heat
exchanging fluid 41 is then transported away, i.e. out of the primary heat exchanger
5 into the tubing of the heat exchanging fluid circulation system 8 by means of the
fluid pump 82. Here, the temperatures are monitored by the control unit 7, processed,
and, in line with a respective algorithm, the control unit 7 drives the fluid pump
82 in order to increase or decrease the heat exchanging fluid flow speed in the heat
exchanging fluid circulation system 8. Thereby, heat can be removed from the primary
heat exchanger 5 faster or slower, as desired, in order to control the heat removal
from the heat pumps 3 arranged in between the mount 2 and the primary heat exchanger
5. Accordingly, the heat is removed from the primary heat exchanger 5 and transferred
into the heat exchanging fluid 41 which, then, is transferred by means of the fluid
pump 82 and the heat exchanging fluid circulation system 8 to the secondary heat exchanger
6, which can transfer the removed heat to ambient air or laboratory infrastructure.
[0047] As an alternative to the previously described device 1 based on fig. 2, fig. 3 depicts
the concept of another embodiment of the device 1' in a structural manner. Accordingly,
it can be gathered from fig. 3 that the device 1' comprises a mount 2' with a structure
in its upper surface, wherein the mount's upper surface comprises indentations 2 which
can be used as wells for receiving the biological samples to be thermocycled, or for
receiving a multi-well plate comprising a plurality of wells comprising the biological
samples therein, wherein several heat pumps 3, such as Peltier elements, are provided
in between the mount 2' and the primary heat exchanger 5' in the form of a solid-to-liquid
heat exchanger. Here, additionally to the respective version in the previously described
embodiment, the primary heat exchanger 5' comprises an inner space 51' including fins
511', such as pin fins or swage fins, within the inner space 51', for example protruding
from one large inner surface side of the inner space 51', the fins 511' providing
a more effective heat transfer from the solid parts of the primary heat exchanger
5' to the heat exchanging fluid 41 perfusing the inner space 51' of the primary heat
exchanger 5, i.e. flowing or being streamed through the inner space 51' of the primary
heat exchanger 5'. Here, a high thermal capacity of the fins 511' allows for a smaller
heat transfer surface and, therefore, for a miniaturized design of the primary heat
exchanger 5'.
[0048] Similarly to the previously described embodiment, a comprehensive heating/cooling
of the mount 2' can be achieved by the structure of device 1', wherein any thermal
connection between the heat pumps 3 and the mount 2' as well as between the heat pumps
3 and the primary heat exchanger 5' is amplified by the use of thermal interface material
31, in order to enhance any temperature conductivity between the mount 2', the heat
pumps 3 and the primary heat exchanger 5'. These components again constituting a thermal
block unit, wherein the components of the thermal block unit, i.e. the mount 2', the
primary heat exchanger 5' and the heat pumps 3 arranged therebetween can be mechanically
connected together, for example by means of a mechanical interface mount in the form
of screws 52', or alternatively threaded bolts, or the like, in combination with a
respective counterpart in the mount, such as a threaded hole, as well as a spring
or a nut-spring combination, connecting and pulling/pushing the primary heat exchanger
5' and the mount 2' together, thereby sandwiching the heat pumps 3 thereinbetween,
i.e. clamping these components together, in order to establish a tight thermally conducting
assembly. For example, the screws 52' can pass through one or several through holes
53' provided in the primary heat exchanger 5' and fit into respective threaded holes
provided in the mount 2'. Further, and similar to the concept as shown in fig. 3,
it can be clearly taken that a heat sink 4' consists of the primary heat exchanger
5' and a secondary heat exchanger 6', wherein the secondary heat exchanger 6', can
be arranged in a spaced-away manner distant from the primary heat exchanger 5', and
wherein the secondary heat exchanger 6' supporting a temperature compensation function
of the primary heat exchanger 5' in regard to the mount 2' can be arranged far away
from the thermal block unit in order to be thermally independent therefrom.
[0049] As can also be gathered from fig. 3, the primary heat exchanger 5' is in fluid connection
with the secondary heat exchanger 6' by means of a heat exchanging fluid circulation
system 8 comprising a fluid pump 82' arranged within the fluid connection between
the primary heat exchanger 5' and the secondary heat exchanger 6', which achieves
the same function as described in regard to the previously described embodiment. In
the presently described embodiment as depicted in fig, 3, the secondary heat exchanger
6' not only allows a streaming of the heat exchanging fluid 41 there through in order
to achieve a fluid-to-solid temperature transfer, but also further comprises fins
61' on its outer surface, such as pin fins or swage fins, for example protruding from
an outer surface of the secondary heat exchanger 6' to the outside, which fins 61'
providing a more effective heat transfer from the solid parts of the primary heat
exchanger 6' to the ambient air. In order to further improve the heat transfer from
the heat exchanging fluid 41 streaming through the secondary heat exchanger 6' to
the solid parts of the secondary heat exchanger 6' itself, to the fins 61' and then
to the ambient air, a fan 62' is provided in connection with the secondary heat exchanger
6' as depicted in fig. 3. Accordingly, in case the temperature of the heated-up heat
exchanging fluid 41 inside the secondary heat exchanger 6' is too high, the fan can
be activated and its speed, i.e. the fan speed, can not only be monitored by the control
unit 7 but can also be increased or decreased, as desired, in order to improve a cooling
performance of the secondary heat exchanger 6'.
[0050] In fig. 4, a modified or complemented embodiment of the device 1' for thermocycling
the biological samples as described in regard to fig. 3 is shown, which is almost
identical but with the difference that the modified device 1' comprises additional
fluid reservoirs 81 within the heat exchanging fluid circulation system 8. In particular,
one fluid reservoir 81 is provided in fluidic connection in between the inner space
51' of the primary heat exchanger 5' and the fluid pump 82, and another fluid reservoir
81 is provided between the inner space 51' of the primary heat exchanger 5' and the
secondary heat exchanger 6' on the other side of the circulation circle. Each fluid
reservoir 81 can accommodate heat exchanging fluid 41 of a predetermined temperature,
which fluid can be provided as part of the circulation system 8, i.e. is fluid-connected
with the circulation system 8, or better with the tubing of the circulation system
8. Thus, the additional fluid in the fluid reservoirs 81 can be used, on demand, in
case heat exchanging fluid 41 of a certain temperature as stored inside the respective
fluid reservoir 81 can be used to heat or cool the heat exchanging fluid 41 streaming
inside the circulation system 8. For example, one of the depicted fluid reservoirs
81 can store cold heat exchanging fluid 41, and the other can store heated heat exchanging
fluid 41, and the control unit 7 can activate the one or the other fluid reservoir
81 in case the heat exchanging fluid 41 inside the tubing of the system 8 must be
cooled down or heated up, by adding the respective cold or hot heat exchanging fluid
41 from one of the reservoirs 81. Thus, depending on the need, the circulation system
8 can comprise multiple hot and/or cold fluid reservoirs 81 accommodating heat exchanging
fluid 41 of different predetermined temperatures, thereby enabling a certain control
of the temperature of the heat exchanging fluid 41 provided to the secondary heat
exchanger 6' or returning to the primary heat exchanger 5'.
[0051] In fig. 5, the depicted flowchart shows the main steps of a method using the inventive
device 1, 1' of the present invention. In particular, the method for thermocycling
biological samples using the device 1, 1' is based on the measuring values as received
from the sensors 9. Here, the method monitors in step 100 the temperature or other
values as received from the sensors 9, and it is determined, in the same step 100,
for example based on the values from the flow sensor 92 or the temperature sensors
91 that the temperature at the mount 2, 2', i.e. the temperature at the samples to
be cycled, is too high or is in need of cooling down. Depending on the outcome of
step 100, the control unit 7 proceeds to step 200, i.e. a step of controlling the
output of the heat pumps 3, the output of the heaters 3', and/or the heat exchange
rate of the primary heat exchanger 5, 5'. Here, for example, the control unit 7 can
increase the fluid pump's performance, or can stop the pumping at all or reverse the
pumping direction, and, optionally, can introduce cold heat exchanging fluid 41 from
the fluid reservoirs 81. Alternatively or additionally, the control unit 7 can increase/decrease
the output of the heaters 3', as desired, and/or can activate or increase the function
of the fan 61' in case the temperature of the primary heat exchanger 5, 5' needs to
be decreased as fast as possible. For example, in step 300, the control unit 7 can
stop or reverse the flow of heat exchanging fluid 41 through the inner space 51, 51'
of the primary heat exchanger 5, 5'. Moreover, in step 400, the control unit 7 can
open one or several of the fluid reservoirs 81 in order to add heated or cooled heat
exchanging fluid 41.
[0052] While the current invention has been described in relation to its specific embodiments,
it is to be understood that this description is for illustrative purposes only. Accordingly,
it is intended that the invention be limited only by the scope of the claims appended
hereto.
1. A device (1; 1') for thermocycling biological samples, the device comprising the following
components:
a mount (2; 2') for receiving said biological samples;
a heat pump (3) for heating and cooling the mount, wherein the heat pump (3) is thermally
coupled to the mount;
a heat sink (4; 4') comprising a primary heat exchanger (5; 5') with an inner space
(51; 51') perfused by a heat exchanging fluid (41), and a secondary heat exchanger
(6; 6'), wherein the primary heat exchanger (5; 5') is thermally coupled to the heat
pump (3), and wherein the secondary heat exchanger (6; 6') is thermally coupled to
the heat pump (3) through the primary heat exchanger (5; 5'); and
a control unit (7) for controlling the thermocycling of said biological samples.
2. The device (1; 1') according to claim 1, wherein the mount (2; 2') is connected to
the heat pump (3) and/or the heat pump (3) is connected to the primary heat exchanger
(5; 5'), preferably by means of a releasable force-fit connection, further preferably
by clamping.
3. The device (1; 1') according to claim 1 or 2, wherein the secondary heat exchanger
(6; 6') is spaced apart from the first heat exchanger (5; 5').
4. The device (1; 1') according to any one of the preceding claims, wherein the secondary
heat exchanger (6; 6') is thermally connected to the primary heat exchanger (5; 5')
by means of a heat exchanging fluid circulation system (8) for circulating said heat
exchanging fluid (41) between the primary heat exchanger (5; 5') and the secondary
heat exchanger (6; 6').
5. The device (1; 1') according to any claim 4, wherein the circulation system (8) comprises
a tubing connection between the primary heat exchanger (5; 5') and the secondary heat
exchanger (6; 6') for circulatory transfer of said heat exchanging fluid (41) between
the primary heat exchanger (5; 5') and the secondary heat exchanger (6; 6'), preferably
wherein the tubing connection is established by flexible tubing for variably positioning
the secondary heat exchanger (6; 6') in relation to the primary heat exchanger (5;
5'); and/or
at least one fluid reservoir (81) accommodating heat exchanging fluid (41) of a predetermined
temperature, preferably wherein the circulation system comprises a plurality of fluid
reservoirs accommodating heat exchanging fluid (41) of different predetermined temperatures.
6. The device (1; 1') according to any one of the preceding claims, wherein at least
one fluid pump (82) is connected to the primary heat exchanger (5; 5') and the secondary
heat exchanger (6; 6') for pumping said heat exchanging fluid (41) and controlling
the flow speed of heat exchanging fluid (41) in the heat sink (4; 4'), preferably
wherein the fluid pump (82) is arranged between the primary heat exchanger (5; 5')
and the secondary heat exchanger (6; 6').
7. The device (1; 1') according to any one of the preceding claims, wherein a thermal
coupling between the mount (2; 2') and the heat pump (3) and/or between the heat pump
(3) and the primary heat exchanger (5; 5') is established by means of a thermal interface
material (31), preferably wherein the thermal interface material (31) comprises at
least one of a carbon based material, such as graphite, preferably pyrolytic graphite,
or graphene, a silicone compound, a phase change material and thermal grease.
8. The device (1; 1') according to any one of the preceding claims, wherein
the primary heat exchanger (5; 5') is a solid-to-liquid heat exchanger, preferably
a compact liquid heat exchanger;
the inner space (51') of the primary heat exchanger (5') comprises fins (511'), preferably
wherein the fins (511') are pin fins or swage fins;
the primary heat exchanger (5; 5') consists of one or more parts made from copper
or other thermally conducting materials; and/or
the primary heat exchanger (5; 5') comprises at least one through hole allowing an
assembly of the device (1; 1') in the manner of a thermally sandwiched structure.
9. The device (1; 1') according to any one of the preceding claims, wherein
the secondary heat exchanger (6; 6') comprises a temperature sensor (91) for controlling
its heat exchange rate;
the secondary heat exchanger (6; 6') comprises an inner space containing heat exchanging
fluid (41) pumped through its inner space, preferably wherein a heat exchange rate
of the secondary heat exchanger (6; 6') is controlled by means of a pump speed sensor
and/or volume flow sensor (92);
the secondary heat exchanger (6; 6') interacts with a working fluid for cooling the
secondary heat exchanger (6; 6'), wherein the heat exchange rate of the secondary
heat exchanger (6; 6') is controlled by means of a working fluid pump; and/or
the secondary heat exchanger (6') interacts with a fan (62') and/or a radiator, wherein
the heat exchange rate of the secondary heat exchanger (6') is controlled by means
of a fan speed sensor and/or a radiator temperature sensor.
10. The device (1; 1') according to any one of the preceding claims, wherein
the mount (2; 2') is made of solid material, preferably silver or aluminum, and functions
as a heat conductor for achieving thermal homogeneity ; and/or
the mount is made of solid thermal conductive material, preferably copper, and comprises
a vapor chamber for transporting and distributing heat between the heat pump (3) and
the biological samples in a uniform manner.
11. The device (1; 1') according to any one of the preceding claims, wherein
the heat pump (3) is one of a thermoelectric device, a resistive heater, or a combination
thereof, the thermoelectric device preferably being a Peltier element, and/or the
resistive heater preferably being one of a ceramic heater, a heating foil, a heating
cartridge or a wire wound heater;
the heat pump (3) is arranged between the mount (2; 2') and the primary heat exchanger,
preferably in a sandwiched manner; and/or
the heat pump (3) is arranged at least in part inside a recess provided in the mount
(2; 2').
12. The device (1; 1') according to any one of the preceding claims, further comprising
at least one sensor (9) for controlling the temperature at the biological samples
received in the mount (2; 2'), preferably wherein the at least one sensor (9) is one
of a temperature sensor (91), a fan speed sensor, a pump speed sensor and a volume
flow sensor (92), further preferably wherein the at least one sensor (9) is provided
at the mount (2; 2'), the heat pump (3), the primary heat exchanger (5; 5'), and/or
the secondary heat exchanger (6; 6').
13. The device (1; 1') according to any one of the preceding claims, wherein the heat
exchanging fluid (41) is a heat exchanging liquid (41), preferably wherein the heat
exchanging liquid (41) is a water-based cooling liquid medium.
14. The device (1; 1') according to anyone of the preceding claims, wherein
said biological samples are placed on the mount (2; 2'), preferably wherein the mount
(2; 2') comprises at least one flat surface (21) or a surface geometry with a plurality
of indentations (22');
said biological samples are received in an array of reaction vessels of a multi-well
plate mounted in a respective recess structure of the mount (2') or on top of the
mount (2); and/or
said biological samples are received in a cartridge or a slide arranged on the mount
(2).
15. The device (1; 1') according to any one of the preceding claims, wherein the device
(1; 1') is a thermocycler for the simultaneous thermocycling of multiple samples to
perform multiple nucleic acid amplification reactions.
16. An instrument for simultaneously monitoring multiple nucleic acid amplification reactions
during thermocycling biological samples, the instrument comprising
a device (1; 1') for thermocycling said biological samples according to any one of
the preceding claims;
at least one excitation light source for applying excitation energy to the nucleic
acid amplification reactions; and
at least one sensor for simultaneously detecting light emitted from the multiple nucleic
acid amplifications.
17. A method for thermocycling biological samples using the device (1; 1') according to
any one of claims 1 to 15, the method comprising the steps of
a) monitoring the temperature at the biological samples; and
b) based on the monitored temperature at the biological samples, controlling the output
of the heat pump (3) and/or the heat exchange rate of the primary heat exchanger (5;
5').
18. The method according to claim 17, wherein the controlling of the heat exchange rate
of the primary heat exchanger (5; 5') in step b) is executed by the control unit (7)
and preferably comprises a control of the heat exchange rate of the second heat exchanger
(6; 6') and/or a stop or reverse of a flow of heat exchanging fluid (41) through the
inner space (51; 51') of the primary heat exchanger (5; 5').
19. The method according to claim 17 or 18, wherein the controlling of the heat exchange
rate of the primary heat exchanger (5; 5') in step b) includes the control of fluid
volume speed to cool the heat exchange fluid (41) in the heat sink (4; 4') above ambient
temperature and approximate to the predetermined lowest temperature of the thermocycles
controlled by the control unit (7).