[0001] The invention refers to a device comprising an inlet channel, outlet channel, first
channel having a hydrodynamic resistance R1, second channel having a hydrodynamic
resistance R2, third channel having a hydrodynamic resistance R3, fourth channel having
a hydrodynamic resistance R4, intermediate channel having a hydrodynamic resistance
R0 and a positioner for positioning an object within the intermediate channel, the
positioner preferably comprising a narrowing of the cross section of the intermediate
channel; the device further comprising an inlet node, outlet node, first intermediate
node and second intermediate node; wherein the inlet channel, second channel and fourth
channel are interconnected via the inlet node, the outlet channel, first channel and
third channel are interconnected via the outlet node, the first channel, second channel
and intermediate channel are interconnected via the first intermediate node and the
third channel, fourth channel and intermediate channel are interconnected via the
second intermediate node; the device further comprising a first valve arranged at
the second channel and configured to be actuated in order to control the flow rate
through the second channel. Furthermore, the invention refers to a system comprising
at least two such devices and a method for operating such system.
[0002] Such a device (and system) is known from
WO 2019 048 713 A1. In particular, it has the purpose to treat and transport an object like a cell or
particle through the device by means of a fluid. In order to control the fluid flow
and fluid direction through the intermediate channel, the device according to
WO 2019 048 713 A1 uses a second valve arranged at the third channel to control the flow rate through
the third channel and furthermore the channels have different resistances. The first
channel has a higher resistance than the third channel and the fourth channel has
a higher resistance than the second channel. To achieve different resistances (i.e.
value of resistance) it is in principle possible to change the cross-section of an
appropriate channel. However, usually the size of cross-section is limited by the
size of objects intended for transport through the channels. In terms of
WO 2019 048 713 A1, an object that is removed from the intermediate channel has to pass the first channel
to enter the outlet node. Thus, the resistance of the first channel can only be varied
by adjusting its length as the size of the cross-section is given by the object size.
Therefore, for several applications it is only possible to vary the resistance of
a channel by varying its length. A longer channel causes more resistance (i.e. loss
of fluid pressure) than a shorter channel. Therefore, it is necessary that the first
channel is longer than the third channel and the fourth channel is longer than the
second channel. This, however, requires a large footprint. This is disadvantageous
since the environments in which such devices are usually used are limited by space
or standards like a 96-well-plate for example. Therefore, the prior art device allows
a relatively small amount of devices being integrated into such biotechnological environments.
[0003] Thus, the technical problem is to provide a device which allows controlling the fluid
flow and fluid direction through the intermediate channel and additionally requires
less space.
[0004] The problem is solved by a device according to claim 1, the system according to claim
9, and a method according to claim 13. Advantageous embodiments are disclosed by the
dependent claims, the description and the figures.
[0005] The basic idea of the invention is to provide a second valve which is arranged at
the fourth channel and configured to be actuated in order to control the flow rate
through the fourth channel. By shifting the second valve from the third to the fourth
channel, it is possible to decrease the size of the channels. For example, the channels
might have the same (small) size or at least the difference between the sizes is smaller
than that of the prior art. In particular, this advantage has an effect on the system
according to the invention since it comprises at least two and preferably several
such devices. This leads to a system which can comprise more devices on the same footprint
compared to the known system. This means that the device density is higher. Furthermore,
the higher the number of devices is, the higher the space-saving is. This significantly
increases the number of objects that can be analyzed with such a system.
[0006] The device is configured in such a way that by actuating the first and/or second
valve, the flow rate and flow direction of a fluid flowing through the intermediate
channel can be controlled. The term "to actuate a valve" means that the valve is operated.
The actuation of the valve includes the opening and closing of the valve. Preferably,
actuation means closing the valve.
[0007] The device is microfabricated. In particular the dimensions of the device and its
components like the channels and valves have a size of up to 5000, 1000, 500, 100,
80, 50, 10, 1, 0,01 µm.
[0008] According to the invention, the first (second) valve is arranged at the second (fourth)
channel and configured to be actuated in order to control the flow rate through the
second (fourth) channel. In particular, the flow rate is controlled by adjusting the
cross-section of the channel, preferably the cross-section at the location of the
valve. The term "flow rate" also comprises the case in which there is no flow through
the channel. In this case the valve is completely closed and the flow rate is zero.
[0009] The device is configured in such a way that by providing a pressure difference between
inlet channel and outlet channel, a fluid being within the device can flow. To allow
a fluid flow from the inlet channel to the outlet channel a pressure at the inlet
channel is higher than the pressure at the outlet channel. A fluid might be a gas
or a liquid.
[0010] The term "closed" means that the valve is at least partially closed, in particular
completely closed. The term "opened" means that the valve is at least partially opened,
in particular completely opened. The term "partially opened" means that the valve
is more opened than closed. For example, in case of a partially opened valve, the
cross-section size of the channel is closer to the cross-section size of the channel
when the valve is completely opened than when the valve is completely closed. The
term "partially closed" means that the valve is more closed than opened. For example,
in case of a partially closed valve, the cross-section size of the channel is closer
to the cross-section size of the channel when the valve is completely closed than
when the valve is completely opened.
[0011] The term that a valve is arranged at an appropriate channel could for example mean
that the valve is arranged outside the channel. In particular, the valve could generate
a pressure on the channel wall thereby decreasing its cross-section. Such valve and
its functioning is described in
WO 2019 048 713 A1, in particular in claims 1 to 37. Such valve is herewith incorporated into the present
application.
[0012] The circumstance that channels are interconnected via a node means that the channels
are in fluid communication with each other. The node is a branch node and in a certain
way similar to a node in an electrical circuit. In a simplified view (e.g. ignoring
compressibility of fluids) and in analogy to Kirchhoff's first law, the sum of the
fluids flowing into the node is equal to the sum of the fluids flowing out of the
node.
[0013] The positioner is for positioning an object within the intermediate channel. Preferably,
the positioner comprises a narrowing of the cross-section of the intermediate channel
wherein the narrowing is preferably arranged between two sections of the intermediate
channel comprising a wider cross-section. This enables an object which flows with
the fluid into the intermediate channel to be positioned (i.e. trapped) by the narrowing.
In an alternative preferred embodiment, the positioner comprises a widening of the
cross-section of the intermediate channel wherein the widening is preferably arranged
between two sections of the intermediate channel comprising a narrower cross-section.
This enables an elastic object with a larger cross-section as the wider cross-sections
to be squeezed through the wider cross-sections by means of the fluid. The elastic
object can be positioned within the widening when it reaches the widening comprising
a larger cross-section than the object. For example, this can be realized by providing
a stop within the widening or by decreasing the fluid pressure when the object reaches
the widening. In an alternative (however not preferred embodiment) the positioning
function is fulfilled by the intermediate channel itself without an additional structure.
For example, the intermediate channel might be of constant cross-section and the positioning
of an elastic object with a wider cross-section that the one of the intermediate channel
can be realized by squeezing the object into the channel and controlling the fluid
pressure in order to move or stop the squeezed object.
[0014] In a preferred embodiment, the object which can be positioned by a positioner is
a matrix, preferably a hydrogel matrix.. According to one embodiment, the matrices
disclosed herein are preferably spherical, e.g. spherical hydrogel matrices but other
forms may also be applied. In a preferred embodiment, the object comprises a matrix
formed using droplet microfluidics. For example, a flow focusing geometry can be used
for the generation of highly monodisperse droplets having a spherical shape. If the
droplet diameter is larger than the width/height of the microfluidic channel in which
the hydrogel formation may occur, formed matrices have a plug-like shape. In addition,
matrices may be formed by conventional pipetting. Thus, matrix solutions comprising
monomers, pre-polymers, precursors, polymer and/or building blocks for gelation/polymerization/curing
reactions may be pipetted on a 2D surface resulting in the formation of a droplet
having the shape of a spherical segment and/or a hemispherical shape. The shape depends
on the surface tension between the droplet and the surrounding surfaces and may be
adjusted by changing the surface characteristics. In another embodiment, matrix solutions
comprising monomers, pre-polymers, precursors, polymer and/or building blocks for
gelation/polymerization/curing reactions may be pipetted into a geometry having a
pre-defined shape (e.g. a cylindrical geometry). Thus, matrices may assume the shape
of the container containing the matrix solution during matrix formation. According
to one embodiment, the object has a diameter of ≤ 1000 µm, such as ≤ 800 µm, ≤ 600
µm or ≤ 400 µm, preferably ≤ 200 µm, such as 5 µm to 150 µm.
[0015] According to one embodiment, the object comprises a hydrogel matrix, which may be
formed upon the gelation/polymerization/curing of a monomer, pre-polymer, precursor,
polymer and/or building block. In one embodiment, the object, preferably having a
spherical shape, comprises a hydrogel, a polymer or pre-polymer which is selected
from the group comprising polyacrylamide, poly(lactic acid) (PLA), polyglycolide (PGA),
copolymers of PLA and PGA (PLGA), poly(vinyl alcohol) (PVA), polyethylene glycol)
(PEG), poly(ethylene oxide), poly(ethylene oxide)-co-poly(propylene oxide) block copolymers
(poloxamers, meroxapols), poloxamines, polyanhydrides, polyorthoesters, poly(hydroxyl
acids), polydioxanones, polycarbonates, polyaminocarbonates, poly(vinyl pyrrolidone),
poly(ethyl oxazoline), carboxymethyl cellulose, hydroxyalkylated celluloses such as
hydroxyethyl cellulose and methylhydroxypropyl cellulose, and natural polymers such
as nucleic acids, polypeptides, polysaccharides, chitosan or carbohydrates such as
polysucrose, hyaluranic acid, dextran and similar derivatives thereof, heparan sulfate,
chondroitin sulfate, heparin, or alginate, and proteins including without limitation
gelatin, collagen, albumin, or ovalbumin, or copolymers, or blends thereof. In particularly
preferred embodiments, the monomers can be selected from polyactic acid) (PLA), poly(vinyl
alcohol) (PVA), polyethylene glycol (PEG) and polyoxazoline (POx). The object preferably
comprises matrix which may comprise polymers and/or precursor molecule, preferably
in a predominantly crosslinked form, which have been disclosed in
PCT/EP2018/074527, in particular, polymers and/or precursor molecules disclosed in claims 101 to 155,
which are herein incorporated by reference.
[0016] In a preferred embodiment, the object comprises a hydrogel. The hydrogel may be a
hydrogel as disclosed in
PCT/EP2018/074527, in particular, hydrogels as disclosed in claims 1 to 51 and 72, which are herein
incorporated by reference.
PCT/EP2018/074527 further discloses methods for producing a hydrogel in claims 52 to 71, which are
herein incorporated by reference. Furthermore, a kit for producing a hydrogel is disclosed
in
PCT/EP2018/074527 in claims 99 and 100, which are also herein incorporated by reference.
[0017] According to a preferred embodiment, the object comprises a hydrogel matrix comprising
one or more selected from the group consisting of a virus particle, a vesicle, such
as an exosome or an apoptotic vesicle, a cell, such as prokaryotic cell, e.g. bacteria,
or a eukaryotic cell and/or a combination thereof. Accordingly, the hydrogel matrix
may contain one or more types and/or sizes of compounds of said group. For instance,
the hydrogel matrix may comprise a cell, such as a eukaryotic cell and a virus. Or
the hydrogel matrix may comprise a vesicle and a cell.
[0018] According to a preferred embodiment, the object comprises at least one cell. Optionally,
the object is a cell. According to a preferred embodiment, the object comprises a
hydrogel matrix comprising at least one cell, also referred to as cell-laden hydrogel
matrix. According to a one embodiment, the object comprises a hydrogel matrix that
provides a three-dimensional environment to the at least one cell, wherein preferably
the matrix is at least 5 µm and ≤ 200 µm in diameter.
[0019] According to a preferred embodiment, the object comprises a hydrogel matrix, preferably
having a spherical shape, comprising more than one cell. According to another embodiment,
the hydrogel matrix comprises a colony of cells. Preferably, a colony of cells can
be located inside the three-dimensional matrix. According to another embodiment, the
cell number changes throughout performing the method. For instance, the cell number
increases over the course of cultivation, decreases over the course of cultivation
or remains constant over the course of cultivation. A colony of cells may be formed
by proliferation of one or more cells, wherein preferably cells proliferate inside
the three-dimensional matrix. In another embodiment, the hydrogel matrix comprises
at least two different types of cells that interact. In particular, the hydrogel matrix
may comprise two different types of cells that interact.
[0020] The at least one cell may be selected from a prokaryotic and/or an eukaryotic cell.
The at least one cell may be selected from the groups consisting of bacteria, archaea,
plants, animals, fungi, slime moulds, protozoa, and algae. According to a preferred
embodiment, the at least one cell may be selected from animal cells, preferably human
cells. According to one embodiment, the at least one cell may be selected from cell
culture cell lines. According to another embodiment, the at least one cells may be
selected from the group consisting of stem cells, bone cells, blood cells, muscle
cells, fat cells, skin cells, nerve cells, endothelial cells, sex cells, pancreatic
cells, and cancer cells. According to another embodiment, the at least one cell may
be derived from cells of the nervous system, the immune system, the urinary system,
the respiratory system, the hepatopancreatic-biliary system, the gastrointestinal
system, the skin system, the cardiovascular system, developmental biology (including
stem cells), pediatrics, organoids, and model organisms. According to another embodiment,
the at least one cell may be derived from one or more of blood and immune system cells,
including erythrocytes, megakaryocytes, platelets, monocytes, connective tissue macrophages,
epidermal Langerhans cells, osteoclast (in bone), dendritic cells, microglial cells,
neutrophil granulocytes, eosinophil granulocytes, basophil granulocytes, hybridoma
cells, mast cells, helper T cells, suppressor T cells, cytotoxic T cells, natural
killer T cells, B cells, natural killer cells, reticulocytes, hematopoietic stem cells,
and committed progenitors for the blood and immune system.
[0021] Preferably, the second (forth) channel is connected to the inlet node with his first
end and connected to the first (second) intermediate node with his second end. Preferably,
the first (third) channel is connected to the first (second) intermediate node with
his first end and connected to the outlet node with his second end. Preferably, the
intermediate channel is connected to the first intermediate node with his first end
and connected to the second intermediate node with his second end.
[0022] In a preferred embodiment the resistances, channels and valves are configured in
such a way that no fluid can flow through the intermediate channel if the valves are
opened. In analogy to the Wheatstone bridge in the field of electrical engineering,
this state can be achieved by fulfilling the equation R1/R2 = R3/R4.
[0023] In a preferred embodiment the resistances, channels and valves are configured in
such a way that a fluid can flow through the intermediate channel from the first intermediate
node to the second intermediate node if the valves are opened. Thereby, the fluid
flows mainly from the second channel to the third channel via the intermediate channel.
This has the advantage that an object entered the intermediate node form the first
intermediate node and then being trapped within the positioner remains safely within
the positioner due to the fluid flow pushing it into the positioner.
[0024] In a preferred embodiment the resistances, channels and valves are configured in
such a way that a fluid can flow through the intermediate channel from the first intermediate
node to the second intermediate node if the first valve is opened and the second valve
is closed. Thereby, the fluid flows mainly from the second channel to the third channel
via the intermediate channel.
[0025] In a preferred embodiment the resistances, channels and valves are configured in
such a way that a fluid can flow through the intermediate channel from the second
intermediate node to the first intermediate node if the second valve is opened and
the first valve is closed. Thereby, the fluid flows mainly from the fourth channel
to the first channel via the intermediate channel. This fluid flow allows releasing
an object, which entered the intermediate channel form the first intermediate node
and is trapped within the positioner, from the positioner and transport it by the
fluid flow to the outlet channel via the first channel.
[0026] In a preferred embodiment the resistances, channels and valves are configured in
such a way that no fluid can flow through the intermediate channel if the valves are
completely closed.
[0027] In one embodiment R1 and R3 are equal or deviate from each other by at most 5%. Additionally,
or alternatively, R2 and R4 are equal or deviate from each other by at most 5%. In
a preferred embodiment R1, R2, R3 and R4 are equal or at least deviate from each other
by at most 5%.
[0028] In one embodiment the first channel and the third channel have the equal geometry.
Additionally or alternatively the second channel and the fourth channel have the equal
geometry. In a preferred embodiment the first to fourth channel have the same geometry.
In particular, the term "same geometry" means the same size of cross-section, the
same shape of cross-section and the same length of channel. The shape of cross-section
is preferably rectangular. Alternative shapes are conceivable like circular for example.
[0029] The invention also refers to a system comprising a first device according to the
invention and at least a second device according to the invention, wherein the devices
are interconnected. In a preferred embodiment the system comprises several devices
according to the invention. In particular, the devices are interconnected in such
a way that they form an array.
[0030] In a preferred embodiment, the system further comprising a feeding line which is
formed by a feeding inlet channel, the intermediate channel of the first device, at
least one connection channel, the intermediate channel of the at least second device
and a feeding outlet channel; wherein the feeding inlet channel, the first channel,
the second channel and the intermediate channel are interconnected via the first intermediate
node; wherein the connection channel, the third channel, the fourth channel and the
intermediate channel are interconnected via the second intermediate node and the connection
channel, the first channel of the second device, the second channel of the second
device and the intermediate channel of the second device are interconnected via the
first intermediate node of the second device; wherein the feeding outlet channel,
the third channel of the second device, the fourth channel of the second device and
the intermediate channel of the second device are interconnected via the second intermediate
node of the second device; wherein the system further comprising a feeding inlet valve
arranged at the feeding inlet channel and configured to be actuated in order to control
the flow rate through the feeding inlet channel; wherein the system further comprising
a feeding outlet valve arranged at the feeding outlet channel and configured to be
actuated in order to control the flow rate through the feeding outlet channel; wherein
the system further comprises a connection valve arranged at the connection channel
and configured to be actuated in order to control the flow rate through the connection
channel.
[0031] This embodiment comprises the preferred case in which more than two devices are interconnected.
(The system comprises "at least" a second device.) The interconnected devices form
an array. For example, if the system comprises three devices, the second device and
the third device are interconnected according to interconnection of the first and
the second device. The feeding line is additionally formed by a second connection
channel wherein the second connection channel, the third channel of the second device,
the fourth channel of the second device and the intermediate channel of the second
device are interconnected via the second intermediate node of the second device and
the second connection channel, the first channel of the third device, the second channel
of the third device and the intermediate channel of the third device are interconnected
via the first intermediate node of the third device. The feeding outlet channel, the
third channel of the third device, the fourth channel of the third device and the
intermediate channel of the third device are interconnected via the second intermediate
node of the third device. In particular, the system further comprises a second connection
valve arranged at the second connection channel and configured to be actuated in order
to control the flow rate through the second connection channel.
[0032] In particular, by completely closing a connection valve it is possible to disconnect
one device from the other as is described in more detail below. This enables to control
the fluid flow within the individual devices independently from each other.
[0033] Preferably, the connection channel is connected to the second intermediate node (of
the first device) with his first end and connected to the first intermediate node
of the second device with his second end. The same applies mutatis mutandis to an
additional connection channel.
[0034] In a preferred embodiment the system preferably comprises a first outer valve configured
to be actuated in order to control the flow rate through the inlet channel of the
first device and the inlet channel of the second device, thereby preferably applying
the same pressure to each inlet channel; further comprising a second outer valve configured
to be actuated in order to control the flow rate through the outlet channel of the
first device and the outlet channel of the second device, thereby preferably applying
the same pressure to each outlet channel. This has the advantage that by activating
one single valve, the flow rate through several channels can be controlled simultaneously.
In particular, by generating a pressure difference between the inlet channels by means
of the first outer valve and the outlet channels by means of the second outer valve
a fluid flow can be induced through several devices simultaneously.
[0035] In an alternative, however also preferred embodiment, the first outer valve is not
provided and/or all first and second valves are actuated in order to control or block
the flow control through the appropriate inlet channels.
[0036] In a preferred embodiment the array formed by the interconnected devices comprises
n times m devices, wherein the devices are arranged in such a way that they from a
matrix of m columns and n rows, wherein m is a number, preferably 16, and n is a number,
preferably 24, wherein the system is preferably configured in such a way that all
first valves of the devices arranged in the same column can be actuated simultaneously
and all first valves arranged in the same row can be actuated simultaneously and/or
all second valves of the devices arranged in the same column can be actuated simultaneously
and all second valves arranged in the same row can be actuated simultaneously.The
last device of a column and the first device of the next column are interconnected
like the first device and the second device of the same column for example. However,
if necessary, a longer connection channel might be used. This preferred embodiment
allows choosing easily the valves of the device intended to be operated.
[0037] A 16x24-matrix is suitable to be integrated into the footprint of a conventional
glass slide (26 x 76 mm) for example. Preferably, the footprint of such matrix is
50 mm times 16 mm (= 800 mm
2). In a further preferred embodiment, n is 32 and m is also 32 so that in total there
are 1024 devices. In a further preferred embodiment, m is 16 and n is 8. Preferably,
the footprint of such matrix is 15 mm times 17 mm (= 255 mm
2).
[0038] The invention also refers to a method comprising the steps: providing a system according
to the invention with the first valve and the second valve of the first device are
closed, the first valve and the second valve of the second device are closed, the
second outer valve is closed, whereas the feeding inlet valve, the connection valve
and the feeding outlet valve are opened; causing a fluid flow within the feeding line
from the feeding inlet channel to the feeding outlet channel; providing an object
within the feeding inlet channel; transporting the object from the feeding inlet channel
within the feeding line by the flow of the fluid.
[0039] In particular, a fluid flow within the feeding line from the feeding inlet channel
to the feeding outlet channel is caused by a pressure difference between the feeding
inlet channel and the feeding outlet channel. The object is for example a cell or
particle or a cell-laden spherical hydrogel matrix. In particular, the object flows
into the intermediate channel and is stopped in the positioner and preferably trapped
by its narrowing or widening of the cross-section.
[0040] In a preferred embodiment the object is trapped by the positioner and the method
comprises the further steps: providing a second object within the feeding inlet channel;
transporting the object from the feeding inlet channel to the positioner of the second
device, wherein the positioner of the first device comprises a structure allowing
the second object passing it while the first object is trapped, the structure being
preferably a bypass. For example, if a first object is trapped by the positioner by
the narrowing of the cross-section, the second object would be led through the bypass
allowing the second object to continue its flow to the intermediate channel of the
second device in order to be trapped in the positioner of the second device. Therewith,
it is possible to "feed" the positioners of all devices of an array with an object
by means of a single feeding line. In a preferred embodiment, the first channel and
the third channel can be used as bypass. If the positioner is occupied by a first
particle, the flow resistance in the intermediate channel rises and a second particle
entering the device through the feeding inlet node can flow through the first channel
and third channel due to a less resistance value of this path. In particular, this
can be achieved by closing the first valve and the second valve as well as closing
the outlet valve.
[0041] The invention also refers to a method comprising the steps: providing a system according
to the invention with the feeding inlet valve, the (or each) connection valve and
the feeding outlet valve are closed, the second outer valve is opened, comprising
the following steps: actuating the first valve of all devices of column x simultaneously,
wherein x is a (natural) number of a range from 1 to m, and/or actuating the second
valve of all devices of row y simultaneously, wherein y is a (natural) number of a
range from 1 to n in order to control the flow rate and flow direction of a fluid
within the intermediate channel of the device which is located in column x and row
y. Thereby, a more flexible operation of the system is possible: In particular, the
flow rate and flow direction within the intermediate channel of one device can be
controlled independently of other devices.
[0042] In a preferred embodiment, the method comprising the steps: providing a system according
to the invention with the feeding inlet valve, the (or each) connection valve and
the feeding outlet valve are closed, the first valve of all devices is opened, the
second valve of all devices is opened, comprising the following steps: Closing the
second valve of all devices, except of the devices in column x, and closing the first
valve of all devices in row y, thereby causing a flow from the second intermediate
node to the first intermediate node within the intermediate channel of the device
which is located in column x and row y. In particular, this device is the only device
with such flow direction within the intermediate channel. In particular, this flow
direction is used to transfer an object which is located within the positioner of
the intermediate channel in a common outer channel or to another position of the system.
In a further preferred embodiment, the method additionally comprises the step of closing
the first valve of all devices in row y2, thereby causing a flow from the second intermediate
node to the first intermediate node within the intermediate channel of the device
which is located in column x and row y2, wherein row y2 is a different row than row
y. This embodiment enables to transfer an object which is located within the positioner
of the intermediate channel of the device which is located in column x and row y2
in a common outer channel or to another position of the system. It is preferred to
close the first valve of all devices in row y and the first valve of all devices in
row y2 simultaneously. Thereby, it is possible to transfer several objects simultaneously.
[0043] In a preferred embodiment, the method comprising the steps: providing a system according
to the invention with the feeding inlet valve, the (or each) connection valve and
the feeding outlet valve are closed, the first valve of all devices is opened, the
second valve of all devices is closed, comprising the following steps: Opening the
second valve of all devices in column x and closing the first valve of all devices
in row y, thereby causing a flow from the second intermediate node to the first intermediate
node within the intermediate channel of the device which is located in column x and
row y. In particular, this device is the only device with such flow direction within
the intermediate channel. In particular, this flow direction is used to transfer an
object which is located within the positioner of the intermediate channel in a common
outer channel or to another position of the system.
[0044] The device and system according to the invention can be preferably used for carrying
out biotechnological processes. One example is an incubation process. For example
the device and system can be used for treatment and forming of objects like cells
(e.g. human or plant cells) and particles. In particular, the system can be used for
the cultivation of single or multiple cells of the same or of different type located
within spherical hydrogel matrices.
[0045] The invention is described by the following figures wherein the figures only illustrate
exemplary embodiments of the invention. In particular, the subject matter of the invention
is not limited to these embodiments.
Figure 1A shows an embodiment of the device according to the invention.
Figure 1B shows a modification of the embodiment according to figure 1A.
Figure 2 shows an embodiment of the system according to the invention.
Figure 2A shows the system according to figure 2 with a different valve configuration.
Figures 3A, 3B and 3C show an exemplary embodiment of the device according to the
invention.
Figure 4 is an exemplary system according to the invention.
Figures 5A to 5C show an exemplary embodiment as 4x4-matrix with different valve configurations.
Figure 6 shows a system consisting of three 8x16-matrixes. FPX: Foot print in X direction.
FPY: Foot print in Y direction.
[0046] Figure 1 shows an embodiment of the device 10 according to the invention. The device
10 is a microfabricated device. It comprises an inlet channel IC, outlet channel OC,
first channel C1 having a hydrodynamic resistance R1, second channel C2 having a hydrodynamic
resistance R2, third channel C3 having a hydrodynamic resistance R3, fourth channel
C4 having a hydrodynamic resistance R4 and a intermediate channel 0 having a hydrodynamic
resistance R0. The intermediate channel comprises a positioner. The positioner is
preferably a widening but could also be a narrowing of the cross section of the intermediate
channel 0.
[0047] The device 10 further comprises an inlet node IN, outlet node ON, first intermediate
node IN1 and second intermediate node IN2.
[0048] The inlet channel IC, second channel C2 and fourth channel C4 are interconnected
via the inlet node IN. The outlet channel OC, first channel C1 and third channel C3
are interconnected via the outlet node ON. The first channel C1, second channel C2
and intermediate channel 0 are interconnected via the first intermediate node IN1.
The third channel C3, fourth channel C4 and intermediate channel 0 are interconnected
via the second intermediate node IN2.
[0049] The device 10 comprises a first valve V1 arranged at the second channel C2 and configured
to be actuated in order to control the flow rate through the second channel C2. In
contrast to known devices, the device according to figure 1 comprises a second valve
V2 arranged at the fourth channel C4 and configured to be actuated in order to control
the flow rate through the fourth channel C4. Thereby, it is possible to control the
flow rate of a fluid flowing through the device 10 as flexibly as desired but without
or less geometric constraints for designing the channels. All fluid flow configurations
can be realized in an easy way by actuating the first valve V1 and/or the second valve
V2. For example R1 to R4 may have the same geometry. The skilled person knows how
to choose the different resistances of the channels in combination with the actuation
of the valves in order to create a specific flow rate. For example, the analogy of
the Wheatstone bridge might be helpful. With reference to the present invention, the
following formula of the Wheatstone bridge might be considered:

wherein
U0 is the pressure difference between the first intermediate node IN1 and second intermediate
node IN2 and U is the pressure difference between input node IN and output node ON.
[0050] By actuating the first valve V1, it is in principle possible to change the resistance
for a fluid flowing through the first channel C1. By actuating the second valve V2,
it is in principle possible to change the resistance for a fluid flowing through the
fourth channel C4. Therefore, by defining the channels with certain resistances and
actuating the valves, it is possible to determine U0. For example, if U0 is zero,
no fluid flows through the intermediate channel. For example, if U0 is positive, a
flow from the first intermediate node IN1 to the second intermediate node IN2 takes
place, whereas a negative value represents a flow in the opposite direction.
[0051] Other than in the known devices, it is in particular not necessary to define the
first channel C1 and the third channel C3 with a different geometry, in particular
with a different length. In principle, it is even possible to define the channels
C1 to C4 with the same geometry and nevertheless achieve a fluid flow as desired.
However, it is preferred to design the channels differently.
[0052] Figure 1B shows the embodiment of figure 1 comprising additionally a feeding inlet
channel FIC having a feeding inlet valve which is described in detail below.
[0053] Figure 2 shows an embodiment of the system according to the invention. It comprises
a first device 10, second device 20, third device 30 and fourth device 40 according
to the invention, wherein the devices are interconnected. The system further comprises
a feeding line FL formed by a feeding inlet channel FIC, the intermediate channel
of all devices and a first connection channel CC1, a second connection channel CC2
and a third connection channel CC3 as well as a feeding outlet channel FOC. This exemplary
embodiment is an array of four devices which are interconnected by the interconnection
channels as described above. Moreover, the system comprises a feeding inlet valve
FIV arranged at the feeding inlet channel FIC and configured to be actuated in order
to control the flow rate through the feeding inlet channel FIC. The system further
comprises a feeding outlet valve FOV arranged at the feeding outlet channel FOC and
configured to be actuated in order to control the flow rate through the feeding outlet
channel FOC. The system further comprises a connection valve CC1-V arranged at the
first connection channel, a second connection valve CC2-V arranged at the second connection
channel and a third connection valve CC3-V arranged at the third connection channel.
The embodiment of figure 2 illustrates a 2x2-matrix, where the first and second device
are arranged in the first column and the third and fourth device are arranged in the
second column. The first line is formed by the first and third device, whereas the
second line is formed by the second and fourth device.
[0054] Figure 2 shows also a first outer valve OV1 configured to be actuated in order to
control the flow rate through the inlet channels of the first to fourth device 10
to 40. A second outer valve OV2 is also provide and configured to be actuated in order
to control the flow rate through the outlet channels of the first to fourth device
10 to 40. The valves in the embodiment according to figure 2 are all opened.
[0055] In figure 2A, the system according to figure 2 is shown in such a state that all
first and second valves of the devices and the second outer valve OV2 are completely
closed. Closed valves are marked by dotted rectangles. Thus, a fluid flow from the
inlet channels to the outlet channels is not possible. Moreover, the valves of the
fluid line are opened, i.e. the valves FIV, CC1-V to CC3-V and FOV. This configuration
has the advantage that a fluid can flow through the feeding line from the feeding
inlet channel FIC to the feeding outlet channel FOC. Thereby, it is possible to provide
objects within the feeding inlet channel FIC and provide them to the individual positioners
sequentially.
[0056] Figure 2B shows the system according to figure 2 with a different valve-actuation
configuration. All valves of the feeding line are completely closed. Thereby, the
different devices are disconnected from each other. This has the advantage that the
fluid flow of each device can be generated individually.
[0057] For example, in the first device 10 a fluid flow is established from the fourth channel
4 to the first channel 1 via the intermediate channel. The fluid flows through the
intermediate channel from the second intermediate node to the first intermediate node.
This is possible since the second valve V2 is opened and the first valve V1 is closed.
Therefore, it is possible to release an object which is trapped in the positioner
of the intermediate channel.
[0058] The second device 20 shows a different actuation of valve V1 and valve V2. Here,
both valves are closed whereby no fluid flows through the intermediate channel.
[0059] The third device 30 comprises two opened valves V1 and V2. The fluid flows mainly
form the second channel to the third channel via the intermediate channel. The fluid
flows through the intermediate channel from the first intermediate node to the second
intermediate node.
[0060] The first valve V1 of the fourth device 40 is opened, whereas the second valve V2
is closed. The fluid flows mainly form the second channel to the third channel via
the intermediate channel. The fluid flows through the intermediate channel from the
first intermediate node to the second intermediate node.
[0061] Figures 3A, 3B and 3C show an exemplary embodiment of the device according to the
invention. The reference signs in this figures which are the same as the reference
signs in figures 1A, 1B and 2 refer to the same features as in this figures.
[0062] As shown in figure 3A, the inlet channel IC is a vertical channel and the arrow indicates
the direction of a fluid flowing into the inlet channel. The outlet channel OC has
a vertical section at his firs end and the arrow indicates the direction of a fluid
flowing out of the outlet channel. The second end of the outlet channel OC is connected
to the outlet node ON. Furthermore, an exemplary embodiment of the positioner P is
shown. The positioner P is arranged within the intermediate channel and forms a widening
of this channel. Within the positioner P there is a trapping structure configured
to trap a particle which flows into the intermediate channel through the first intermediate
node FIN. On each side of the trapping structure a bypass channel BC is arranged to
enable a second particle to pass the positioner P and flow to the second intermediate
node IN2 if a first particle is already positioned within the trapping structure of
the positioner P. The feeding inlet channel FIC has a feeding inlet valve FIV which
is a membrane and can be deformed in order to control the flow rate through the feeding
inlet channel FIC. Furthermore, the first valve V1 and the second valve V2 are elastic
elements which can be deformed to control the flow rate through the second channel
C2 and the fourth channel C4. The valves V1 and V2 are equal in shape and size. The
fourth channel comprises two sections. The first section is the valve V2 and the second
section is the part between the inlet node IN and the valve V2. The first channel
C1 is formed by the first valve V2. The height and width of the intermediate channel,
the first channel C1 and the outlet channel OC is almost the same. These channels
are provided to transport a particle of e.g. 80 µm. Compared with the other channels
(e.g. third channel C3) these channels have a larger cross-section. In this embodiment,
the first channel C1 and the third channel C3 have the same resistance. Due to the
fact that the first channel has a larger cross-section, it must be longer to cause
the same pressure loss.
[0063] Figure 3B shows the device of figure 3A in the top view. Figure 3C depicts the device
of figures 3B with a schematically indication of the resistances of the channels C1
to C4. The first channel C1 has the resistance R1 and the third channel C3 has the
resistance R3. The second channel C2 has the total resistance R2* which is the sum
of the resistance R2 of channel C2 and the resistance RiV1 of the first valve V1 in
the opened stated. The resistance of the first valve RiV1 can be changed by closing
the valve. Therefore, RiV1 is a flexible resistance. The same applies mutatis mutandis
to total resistance R4* which is the sum of the resistance R4 of the fourth channel
C4 and the resistance of the second valve RiV2. Furthermore, RiV3 is illustrated which
is the flexible resistance of the feeding inlet valve FIV.
[0064] Figure 4 is an exemplary system according to the invention. In principle it is an
array as depicted in schematically diagram of figure 2. However, the connection channels
connecting the devices 10 to 40 are not illustrated for a better overview. Further,
a common channel COC connecting the outlet channels of the individual devices is shown.
[0065] Figures 5A to 5C show an exemplary embodiment as 4x4-matrix with different valve
configurations. The same pressure must be applied at each inlet. This is guaranteed
by the tree structure of the supply channel. Figure 6 shows a system consisting of
three 8x16-matrixes.
[0066] It is preferred that (all) valves V1 of one column may be operated simultaneously
using one external (solenoid) valve. Additionally or alternatively, (all) valves V2
of one row may be operated simultaneously using on (external) solenoid valve. Thus,
for an exemplary 4x4-matrix (four columns and four rows), eight external valves are
needed. For an exemplary 16x24-matrix (16 columns and 24 rows), 40 external valves
are needed.
[0067] Figure 6 shows an illustration of an exemplary system having 16 x 24 devices. The
length in X direction (FPX) is 50 mm and the length in Y direction (FPY) is 16 mm
resulting in a footprint of 800 mm
2 in total. Thus, the mean footprint of one device is 2.08 mm
2.
1. Device (10) comprising an inlet channel (IC), outlet channel (OC), first channel (C1)
having a hydrodynamic resistance R1, second channel (C2) having a hydrodynamic resistance
R2, third channel (C3) having a hydrodynamic resistance R3, fourth channel (C4) having
a hydrodynamic resistance R4, intermediate channel (0) having a hydrodynamic resistance
R0 and a positioner (P) for positioning an object within the intermediate channel
(0), the positioner (P) preferably comprising a narrowing of the cross section of
the intermediate channel (0); the device (10) further comprising an inlet node (IN),
outlet node (ON), first intermediate node (IN1) and second intermediate node (IN2);
wherein the inlet channel (IC), second channel (C2) and fourth channel (C4) are interconnected
via the inlet node (IN), the outlet channel (OC), first channel (C1) and third channel
(C3) are interconnected via the outlet node (ON), the first channel (C1), second channel
(C2) and intermediate channel (0) are interconnected via the first intermediate node
(IN1) and the third channel (C3), fourth channel (C4) and intermediate channel (0)
are interconnected via the second intermediate node (IN2); the device (10) further
comprising a first valve (V1) arranged at the second channel (C2) and configured to
be actuated in order to control the flow rate through the second channel (C2), characterized in that the device (10) comprises a second valve (V2) arranged at the fourth channel (C4)
and configured to be actuated in order to control the flow rate through the fourth
channel (C4).
2. Device (10) according to the previous claim, wherein the resistances, channels and
valves are configured in such a way that no fluid can flow through the intermediate
channel (0) if the valves (V1, V2) are opened or alternatively the resistances, channels
and valves are configured in such a way that a fluid flows through the intermediate
channel (0) from the first intermediate (IN1) node to the second intermediate node
(IN2) if the valves (V1, V2) are opened.
3. Device (10) according to one of the previous claims, wherein the resistances, channels
and valves are configured in such a way that a fluid can flow through the intermediate
channel (0) from the first intermediate node (IN1) to the second intermediate node
(IN2) if the first valve (V1) is opened and the second valve (V2) is closed.
4. Device (10) according to one of the previous claims, wherein the resistances, channels
and valves are configured in such a way that a fluid can flow through the intermediate
channel (0) from the second intermediate node (IN2) to the first intermediate node
(IN1) if the second valve (V2) is opened and the first valve (V1) is closed.
5. Device (10) according to one of the previous claims, wherein R1 and R3 are equal or
deviate from each other by at most 5% and/or wherein R2 and R4 are equal or deviate
from each other by at most 5%.
6. Device (10) according to one of the previous claims, wherein the first channel (C1)
and the third channel (C3) have the equal geometry and/or wherein the second channel
(C2) and the fourth channel (C4) have the equal geometry.
7. System comprising a first device (10) according to one of the previous claims and
at least a second device (20) according to one of the previous claims, wherein the
devices (10, 20) are interconnected.
8. System according to the previous claim, further comprising a feeding line (FL) which
is formed by a feeding inlet channel (FIC), the intermediate channel (0) of the first
device (C1), at least one connection channel (CC1), the intermediate channel (0) of
the at least second device (20) and a feeding outlet channel (FOC); wherein the feeding
inlet channel (FIC), the first channel (C1), the second channel (C2) and the intermediate
channel (0) are interconnected via the first intermediate node (IN1); wherein the
connection channel (CC1), the third channel (C3), the fourth channel (C4) and the
intermediate channel (0) are interconnected via the second intermediate node (IN2)
and the connection channel (CC1), the first channel (C1) of the second device (20),
the second channel (C2) of the second device (20) and the intermediate channel (0)
of the second device (20) are interconnected via the first intermediate node (IN1)
of the second device (20); wherein the feeding outlet channel (FOC), the third channel
(C3) of the second device (20), the fourth channel (C4) of the second device (20)
and the intermediate channel (0) of the second device (20) are interconnected via
the second intermediate node (IN2) of the second device (20); wherein the system further
comprising a feeding inlet valve (FIV) arranged at the feeding inlet channel (FIC)
and configured to be actuated in order to control the flow rate through the feeding
inlet channel (FIC); wherein the system further comprising a feeding outlet valve
(FOV) arranged at the feeding outlet channel (FOC) and configured to be actuated in
order to control the flow rate through the feeding outlet channel (FOC); wherein the
system further comprises a connection valve (CC1-V) arranged at the connection channel
(CC1) and configured to be actuated in order to control the flow rate through the
connection channel (CC1).
9. System according to the previous claim, further preferably comprising a first outer
valve (OV1) configured to be actuated in order to control the flow rate through the
inlet channel (IC) of the first device (10) and the inlet channel (IC) of the second
device (20), thereby preferably applying the same pressure to each inlet channel (IC);
further comprising a second outer valve (OV2) configured to be actuated in order to
control the flow rate through the outlet channel (OC) of the first device (10) and
the outlet channel (OC) of the second device (20), thereby preferably applying the
same pressure to each outlet channel.
10. System according to one of the previous system claims, wherein the array formed by
the interconnected devices comprises n times m devices, wherein the devices are arranged
in such a way that they form a matrix of m columns and n rows, wherein m is a number,
preferably 16, and n is a number, preferably 24, wherein the system is preferably
configured in such a way that all first valves (V1) of the devices arranged in the
same column can be actuated simultaneously and all first valves (V1) of the devices
arranged in the same row can be actuated simultaneously and/or all second valves (V2)
of the devices arranged in the same column can be actuated simultaneously and all
second valves (V2) of the devices arranged in the same row can be actuated simultaneously.
11. Method comprising the steps: providing a system according to one of the previous system
claims with the first valve (V1) and the second valve (V2) of the first device are
closed, the first valve (V1) and the second valve (V2) of the second device are closed,
the second outer (OV2) valve is closed, whereas the feeding inlet valve (FIV), the
connection valve (CC1) and the feeding outlet valve (FOV) are opened; causing a fluid
flow within the feeding line (FL) from the feeding inlet channel (FIC) to the feeding
outlet channel (FOC); providing an object within the feeding inlet channel (FIC);
transporting the object from the feeding inlet channel (FIC) within the feeding line
by the flow of the fluid.
12. Method according to the previous claim, wherein the object is trapped by the positioner
of the first device (10); further comprising the steps: providing a second object
within the feeding inlet channel (FIC); transporting the object from the feeding inlet
channel (FIC) to the positioner (P) of the second device (20), wherein the positioner
(P) of the first device (10) comprises a structure allowing the second object passing
it, the structure being preferably a bypass.
13. Method according to the previous claims, wherein the first and/or second object has
one or more of the following characteristics:
- the object comprises a matrix, in particular a hydrogel matrix, and preferably has
a spherical shape;
- the object comprises a matrix, preferably a hydrogel matrix, formed using droplet
microfluidics;
- the object comprise a hydrogel matrix formed upon the gelation/polymerization/curing
of a monomer, pre-polymer, precursor, polymer and/or building block;
- the object comprises at least one cell, optionally the object is a cell; and/or
- the object comprises a hydrogel matrix comprising at least one cell, optionally,
∘ the hydrogel matrix comprises one cells;
∘ the hydrogel matrix comprises a colony of cells of the same type;
∘ the hydrogel matrix comprises two or more different types of cells; or
∘ the hydrogel matrix comprises colonies of two or more different types of cells.
14. Method comprising the steps: providing a system according to one of the previous system
claims with the feeding inlet valve (FIV), the connection valve (CC1-V) and the feeding
outlet valve (FOV) are closed, the second outer valve (OV2) is opened, comprising
the following steps: actuating the first valve (V1) of all devices of column x simultaneously,
wherein x is a number of a range from 1 to m, and/or actuating the second valve (V2)
of all devices of row y simultaneously, wherein y is a number of a range from 1 to
n in order to control the flow rate and flow direction of a fluid within the intermediate
channel (0) of the device which is located in column x and row y.