CN215050110U - Suspended array micro-fluidic chip - Google Patents
Suspended array micro-fluidic chip Download PDFInfo
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- CN215050110U CN215050110U CN202120515756.5U CN202120515756U CN215050110U CN 215050110 U CN215050110 U CN 215050110U CN 202120515756 U CN202120515756 U CN 202120515756U CN 215050110 U CN215050110 U CN 215050110U
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Abstract
The utility model provides a suspended array microfluidic chip, which comprises a channel layer and a basal layer; the channel layer is positioned on the substrate layer; the channel layer comprises a sample inlet, a sample inlet channel, a sample chamber, a sample outlet channel and a sample outlet which are connected in sequence, the sample chamber comprises at least two mutually independent shells, and a main channel and a plurality of capture grooves communicated with the main channel are arranged in each shell; the trapping groove structure can effectively promote the cells to preferentially enter the groove, and obtains the single cell trapping efficiency of more than 80 percent; a plurality of connecting channels are arranged between any two adjacent shells, two ends of each connecting channel respectively correspond to one capturing groove and are communicated with the upper parts of the capturing grooves to form a suspended array channel structure, and the study on intercellular communication of single-cell level TNTs can be met.
Description
Technical Field
The utility model relates to a micro-fluidic technical field specifically relates to a unsettled array micro-fluidic chip.
Background
Lab-on-a-chip (Lab on a chip) has been developed as one of the leading science and technology fields in the world today (sens. actors, B, 1990, 1, 244-. The micro-fluidic chip is a technical platform which utilizes micro-electro-mechanical technology to process functional units such as micro-channels, micro-reactors and the like on matrix materials such as glass, high molecular polymers and the like, and realizes integration or basic integration of basic operation units such as pretreatment, sample adding, reaction, separation, analysis, cell culture and the like related to the fields of biology, chemistry and the like on a chip with a square size so as to replace various functions of a conventional chemical or biological laboratory.
The micro-fluidic chip has the characteristics of large specific surface area, high mass and heat transfer rate, low reagent consumption, environmental friendliness, easiness in large-scale integration, high-throughput reaction and the like, and has obvious superiority in the application of the micro-fluidic chip in the fields of water environment pollution, protein analysis, gene analysis, bionic research, cell biology and the like. Among them, in the field of cell biology, microfluidic chips have been used as an excellent tool for studying intercellular communication. Intercellular communication, one of the intercellular interactions of multicellular organisms, plays a crucial role in the growth, development and maintenance of living organisms. Among them, a special plasmodesmata (TNTs) is a kind of connective structure for communication between mammalian cells. Unlike the connection modes of gap connection, chemical synapse and the like which only transmit molecular information, TNTs can also remotely transport vesicles, organelles, viral proteins and the like, promote deep-level communication among cells and even participate in stem cell treatment, and the abnormality of the TNTs can cause the occurrence of tumors and neurodegenerative diseases, so the TNTs have wide application potential in clinical diagnosis and treatment research. However, the existing microfluidic chip still cannot effectively realize the research of exploring TNTs dynamic connection, cell component transfer, cell drug stress and the like on the single cell level.
SUMMERY OF THE UTILITY MODEL
The utility model discloses to the weak point that current micro-fluidic chip shows in the aspect of the technical problem who mentions in solving the background art, designed a micro-fluidic chip with unsettled array channel, satisfied the research demand of unicellular level TNTs cell communication.
The utility model adopts the technical proposal that:
a suspended array microfluidic chip comprises a channel layer and a substrate layer, wherein the channel layer is positioned on the substrate layer;
the channel layer comprises a sample inlet, a sample introduction channel, a sample chamber, a sample outlet channel and a sample outlet which are sequentially connected, the sample chamber comprises at least two mutually independent shells, a main channel and a plurality of capture grooves communicated with the main channel are arranged in each shell, and two ends of each main channel are respectively communicated with the sample introduction channel and the sample outlet channel;
a plurality of connecting channels are arranged between any two adjacent shells, and two ends of each connecting channel respectively correspond to one capturing groove and are communicated with the upper part of the capturing groove.
Further preferably, the sample outlet channel comprises at least one S-shaped bend.
Further preferably, the height dimension of the connecting channel is not more than 10 microns, and the width dimension of the connecting channel is not more than 10 microns; and/or the length of the connecting channel is 5-200 microns.
Further preferably, the height of the main channel and the height of the capture groove are both 30-60 micrometers.
Further preferably, the projection length of the connecting part of the capturing groove and the main channel on the substrate layer is 10-20 microns, and/or the width of the main channel is 50-300 microns.
Further preferably, the distance between the center lines of two adjacent connecting channels is not less than the projected length.
Further preferably, the included angle between the connecting channel and the length direction of the main channel is 45-135 degrees.
Further preferably, the projection shape of the capturing groove on the substrate layer is a fan shape.
Further preferably, the suspended array microfluidic chip further comprises a support layer, and the channel layer is located between the support layer and the substrate layer.
Further preferably, the support layer and the channel layer are made of any one of rigid plastic and glass.
The utility model has the advantages that:
the capture groove structure designed in the sample cavity of the suspended array microfluidic chip provided by the utility model is matched with the S-shaped bent structure of the sample outlet channel, so that cells can be effectively promoted to preferentially enter the capture groove for capture, and the single cell capture efficiency of more than 80 percent can be obtained; the sample chamber comprises a connecting channel structure arranged in a suspended array, the structure is positioned on the upper part of the sample chamber and is specifically communicated with the upper part of a capture groove arranged in the sample chamber, the structure can be adapted to the free suspension state of TNTs, and meanwhile, the size of the connecting channel is smaller than that of a cell body, so that the selectivity of cell types on each side of the connecting channel can be effectively ensured, and finally, the research of intercellular communication of the TNTs at the level of a single cell is met.
Drawings
In order to more clearly illustrate the technical solution of the present invention, the drawings required to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some of the embodiments described in the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without inventive labor.
Fig. 1 is an assembly schematic diagram of the suspended array microfluidic chip in example 1;
FIG. 2 is a schematic plan view of the channel layer in example 1;
FIG. 3 is an enlarged schematic view of a portion of the sample chamber shown in FIG. 2, the enlarged portion being defined by a rectangle in phantom at A;
FIG. 4 is a schematic perspective view of the connecting channels of the suspended array in the sample chamber described in example 1;
FIG. 5 is a schematic cross-sectional view of the connecting channels of the suspended array in the sample chamber described in example 1;
FIG. 6 is a conventional fluorescence microscopy imaging showing single cell capture within a microfluidic chip, wherein the chip architecture is bright field imaging and the cell fluorescence is acquired in a dark field fluorescence mode;
FIG. 7 is a confocal fluorescence microscopy image showing intercellular TNT structure within the dangling array channels and material transport within the TNT;
fig. 8 is a schematic diagram 1 of a cantilever channel inside a confocal 3D imaging display suspended array microfluidic chip;
fig. 9 is a schematic diagram 2 showing cantilever channels inside a suspended array microfluidic chip by confocal 3D imaging;
FIG. 10 is a schematic plan view of a channel layer in example 2;
FIG. 11 is an enlarged schematic view of a portion of the sample chamber shown in FIG. 10, the enlarged portion being defined by the rectangle in phantom at B;
FIG. 12 is a schematic plan view of a channel layer in example 3;
FIG. 13 is an enlarged schematic view of a portion of the sample chamber shown in FIG. 12, the enlarged portion being defined by the rectangle in phantom at C;
fig. 14 is a two-dimensional code address for storing the color original drawings of fig. 4 to 5.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.
It should be noted that all the directional indicators (such as upper, lower, left, right, front and rear … …) in the embodiments of the present invention are only used to explain the relative position relationship between the components, the motion situation, etc. in a specific state (as shown in the drawings), and if the specific state is changed, the directional indicator is changed accordingly. In the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise. In the present application, unless expressly stated or limited otherwise, the terms "connected" and "fixed" are to be construed broadly, e.g., "fixed" may be fixedly connected or detachably connected, or integrally formed; can be mechanically or electrically connected; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise.
Example 1:
a suspended array microfluidic chip, as shown in fig. 1, includes a channel layer 10, a substrate layer 20, and a support layer 30. Wherein the channel layer 10 is located between the base layer 20 and the support layer 30. During preparation, the prepared channel layer 10 is bonded with the support layer 30 to prepare a chip with a closed channel, and then the chip with the closed channel is bonded with the substrate layer 20 to prepare the suspended array microfluidic chip. Wherein holes may be pre-drilled in the support layer 30 to facilitate bonding with the channel layer 10. Preferably, the material of the support layer 30 and the channel layer 10 is PDMS, and in other embodiments, the support layer 30 and the channel layer 10 may be made of PMMA, PET, PBT, or other base materials; the substrate layer 20 may be a clear glass cover slip.
In this embodiment, as shown in fig. 2, the channel layer 10 includes a sample inlet 11, a sample inlet channel 12, a sample chamber 13, a sample outlet channel 14, and a sample outlet 15, which are connected in sequence; specifically, the number of the sample inlets 11, the sample channels 12, the sample outlets 14, and the sample outlets 15 may be multiple, as shown in fig. 2, the channel layer 10 of this embodiment includes two sample inlets 11, two sample channels 12, two sample outlets 14, and two sample outlets 15.
The structure inside the rectangle dashed box at a in fig. 2 is enlarged to obtain the specific structure of the sample chamber 13 in this embodiment, as shown in fig. 3: the sample chamber 13 comprises at least two mutually independent shells, each shell is internally provided with a main channel 131 and a plurality of capture grooves 132 communicated with the main channel 131, and two ends of each main channel 131 are respectively communicated with the sample inlet channel 12 and the sample outlet channel 14; a plurality of connecting channels 133 are provided between any two adjacent housings, and both ends of each connecting channel 133 correspond to one catching groove 132 respectively and communicate with the upper portion of the catching groove 132.
Specifically, in this embodiment, the sample chamber 13 includes two independent housings, each housing has a main channel 131 therein, and the width of the main channel 131 can be adjusted between 50 to 300 micrometers, which is larger than the size of the sample. The size of the sample is on the micron scale, such as cells in the field of cell biology, and in particular, the chip can be used for studying cell-to-cell communication in mammals, but does not exclude carrier chips that can also be used in other fields such as protein analysis, genetic fragment analysis, and the like.
In this embodiment, 50 catching grooves 132 are provided in communication with each other on the side of each main passage 131, and 50 connecting passages 133 are provided in an array between the two housings. All the capturing grooves 132 are disposed on the same side, specifically on one side of the corresponding main channel 131 facing the other main channel, with reference to fig. 3, in this embodiment, the projection shape of the capturing groove 132 on the substrate layer 20 is a semicircle (with a circle center of O and a radius of R), the radius R of the semicircle capturing groove 132 is 10 to 20 micrometers, the capturing grooves with different sizes can be designed according to the sizes of different samples by specific design values, and when the capturing grooves are used for researching intercellular communication, the capturing grooves with different sizes are designed according to the sizes of different cells by the value design, so as to meet the capturing requirement of a single cell. The heights of the main channel 131 and the capture groove 132 can be adjusted between 30-60 microns, and the specific values are adaptively designed according to the size of the cells in the practical application scenario.
With continued reference to FIG. 3, the distance d between the center lines of two adjacent connecting channels 133 is larger than the diameter of the trapping groove 132, i.e., the distance d > 2R, so that good cell trapping efficiency can be ensured. In other preferred embodiments, the distance d may be designed to be 1.5 times the diameter of the capture groove 132, i.e., the distance d is 1.5 × 2R, and the value of the distance d may also be designed to be greater than 1.5 × 2R, which is also within the scope of the present invention, in order to avoid the narrow interval between the connecting channels 133 from affecting the efficiency of the capture groove 132 in capturing cells.
In the above paragraph, "height" in "the height of the main channel 131 and the catching groove 132" refers to channel height. Referring specifically to FIG. 4: the main channel 131, the capturing groove 132 and the connecting channel 133 involved in this embodiment are flat, wherein the channel shape of the main channel 131 and the connecting channel 133 is a rectangular channel, wherein the side parallel to the direction of the sample flow is the length L of the channel, the side perpendicular to the plane of the substrate layer 20 is the height H of the channel, and the other side is the width W of the channel. It should be noted that the description of the length, height and width of each channel referred to in the application text of the present invention is explained in terms of the channel length, channel height and channel width indicated in fig. 4.
Specifically, in the present embodiment, the height dimension of the connecting channel 133 is 10 micrometers, the width dimension is 10 micrometers, and the length dimension is 100 micrometers. In other embodiments, the height and width dimensions of the connecting channel 133 may be not more than 10 μm, so as to obtain a product capable of achieving the technical objects of the present invention. Wherein, this connecting channel 133's length dimension also can be adjusted at 5 ~ 200 microns length within range, this is mainly based on the bionical design that TNTs's structural feature was done the utility model discloses the 5 ~ 200 microns length within range of injecing, the demand of intercellular communication can all be satisfied.
Different from the adherent cell structure, the TNTs are usually in a free suspension state and in dynamic balance of formation, maintenance and regression, and the existing microfluidic chip cannot adapt to the characteristics of the TNTs and is difficult to realize effective research. To solve this problem, the present embodiment designs the following structure: continuing to refer to the perspective view of fig. 4, the height of the connecting channel 133 is less than the height of the main channel 131, and the connecting channel 133 is located at the upper portion of the sample chamber 13, specifically, is communicated with the upper portion of the capturing groove 132 arranged in the sample chamber 13, forming the innovative structure of the present invention: this feature is more visually illustrated by the cross-sectional view of the suspended array channel, sample chamber 13 in fig. 5. The structure is very beneficial to research on the intercellular communication at the single cell level, and is particularly beneficial to be used as a chip carrier for exploring dynamic connection of TNTs.
Specifically, in this embodiment, each sample outlet channel 14 further comprises at least one S-shaped curve (the S-shaped curve is not directly labeled in fig. 2), and with continued reference to fig. 4 and 5, after the cell suspension is added from the sample inlet 11, due to the design of the S-shaped curve of the sample outlet channel 14, the outlet end has a high fluid impedance, so that the cells preferentially enter the capture groove 132 and occupy the space in the groove; meanwhile, the connecting channel 133 is located at the upper portion of the sample chamber 13 and has a size smaller than that of the cell body, so that it can be effectively ensured that the cells do not enter the opposite side, the selectivity of cell types at both sides can be satisfied, and the formation of TNTs can be observed between the cells at both sides. In other embodiments, the height and width of each connecting channel 133 may also be less than 10 μm, and the minimum size may be designed as low as several hundred nanometers, so as to satisfy the requirement of TNTs passing and realizing the communication between cells.
In another embodiment, the distance d between the center lines of two adjacent connecting channels 133 is designed to be equal to the diameter of the capturing groove 132, that is, d is 2R, the design of the sample outlet channel 14 and the connecting channel 133 is the same as above, when the suspended array microfluidic chip designed in this embodiment is used to study the communication between cells, firstly, a micro-injection pump or a vacuum pump is used to perform positive or negative sample injection, and a cell suspension is added from the sample inlet 11 on one side, as shown in fig. 6, cells preferentially enter the capturing groove 132 and do not enter the opposite side, and finally, the capturing efficiency of single cells is more than 80%; after the cells on the sample injection side grow well in an adherent manner, the cells are seeded into the opposite side by the same method, and after 2-3 days, the formation of TNTs can be observed among the cells on the two sides, specifically, as shown in figure 7, mitochondria (GFP and green fluorescent protein) are specifically marked in the cell on one side, and the process that the donor cell transmits to the receptor cell mitochondria through the TNTs is observed by means of a confocal microscope, so that the research requirement of the single cell level TNTs cell communication is met.
Also can use confocal 3D formation of image to show the technique and observe the utility model discloses the inside connecting channel 133 of unsettled array micro-fluidic chip of protection, this connecting channel 133 is the cantilever and sets up between main channel 131, specifically as shown in figure 8.
In another embodiment, the positional setting relationship between the connection channel 133 and the main channel 131 is changed to: the included angle between the connecting channel 133 and the main channel 131 in the length direction is 45 °, and the confocal 3D imaging display result is shown in fig. 9, at this time, the cell capture efficiency of more than 80% can be achieved. In other embodiments, the included angle between the connecting channel 133 and the main channel 131 in the length direction can be adjusted between 45 ° and 135 °, which is within the protection scope of the present invention.
Example 2:
the structure of the channel layer 10 of the suspended array microfluidic chip prepared by this embodiment is slightly different from that of embodiment 1, but all belong to the protection range of the present invention, and can obtain the cell capture efficiency of more than 80%, and meet the research requirements of single cell level TNTs cell communication.
As shown in fig. 10, the channel layer 10 includes a sample inlet 11, a sample channel 12, a sample chamber 13, a sample outlet channel 14 and a sample outlet 15, which are connected in sequence; specifically, the number of the sample inlets 11, the sample channels 12, the sample outlets 14, and the sample outlets 15 may be plural. Specifically, the channel layer 10 of the present embodiment includes two sample inlets 11, two sample channels 12, two sample outlets 14, and two sample outlets 15. Wherein, two introduction ports 11, introduction channel 12 are separately arranged on both sides of the sample chamber 13, and the introduction directions of the two groups of samples are opposite, correspondingly, two exit channels 14 are also separately arranged on both sides of the sample chamber 13, and the exit directions of the samples are also opposite.
The structure inside the rectangle dashed box at B in fig. 10 is enlarged to obtain the specific structure of the sample chamber 13 in this embodiment, as shown in fig. 11: the sample chamber 13 comprises at least two mutually independent shells, each shell is internally provided with a main channel 131 and a plurality of capture grooves 132 communicated with the main channel 131, and two ends of each main channel 131 are respectively communicated with the sample inlet channel 12 and the sample outlet channel 14; a plurality of connecting channels 133 are provided between any two adjacent housings, and both ends of each connecting channel 133 correspond to one catching groove 132 respectively and communicate with the upper portion of the catching groove 132.
In this embodiment, the angle between the connecting channel 133 and the main channel 131 in the longitudinal direction is 45 °, and other technical features are the same as those of embodiment 1.
Example 3:
the structure of the channel layer 10 of the suspended array microfluidic chip prepared by this embodiment is slightly different from that of embodiment 1, but all belong to the protection range of the present invention, and can obtain the cell capture efficiency of more than 80%, and meet the research requirements of single cell level TNTs cell communication.
As shown in fig. 12, the channel layer 10 includes a sample inlet 11, a sample channel 12, a sample chamber 13, a sample outlet channel 14 and a sample outlet 15, which are connected in sequence; specifically, the number of the sample inlets 11, the sample channels 12, the sample outlets 14, and the sample outlets 15 may be plural. Specifically, the channel layer 10 of the present embodiment includes three sample inlets 11, three sample channels 12, three sample outlets 14, and three sample outlets 15.
The structure inside the rectangle dashed box at C in fig. 12 is enlarged to obtain a specific structure of the sample chamber 13 in this embodiment, as shown in fig. 13: the sample chamber 13 comprises at least three mutually independent shells, each shell is internally provided with a main channel 131 and a plurality of capture grooves 132 communicated with the main channel 131, and two ends of each main channel 131 are respectively communicated with the sample inlet channel 12 and the sample outlet channel 14; a plurality of connecting channels 133 arranged in an array are arranged between any two adjacent shells, and two ends of each connecting channel 133 respectively correspond to one capturing groove 132 and are communicated with the upper part of the capturing groove 132.
In other embodiments, according to actual requirements, for example, when cell-to-cell communication is studied, the number of the cells involved is four, five or more, and the number of the sample inlets 11, the sample inlet channels 12, the main channels 131 in the sample chamber 13, the sample outlet channels 14 and the sample outlets 15 can also be set to four, five or more, and these design schemes are all within the protection scope of the present invention.
For more clearly explaining the technical solutions and the technical effects achieved by the embodiments of the present invention, the two-dimensional code of the color original image shown in fig. 4 to 5 is attached to view the address, as shown in fig. 14.
The above mentioned is only the preferred embodiment of the present invention, and the patent scope of the present invention is not limited thereby, all the technical ideas of the present invention are that the various equivalent structures made by the contents of the specification and the drawings are changed or directly/indirectly applied to other related technical fields, and all the equivalent structures should belong to the scope defined in the claims of the present invention as long as they do not depart from the spirit of the present invention.
Claims (10)
1. A suspended array microfluidic chip is characterized in that:
the device comprises a channel layer and a substrate layer, wherein the channel layer is positioned on the substrate layer;
the channel layer comprises a sample inlet, a sample introduction channel, a sample chamber, a sample outlet channel and a sample outlet which are sequentially connected, the sample chamber comprises at least two mutually independent shells, a main channel and a plurality of capture grooves communicated with the main channel are arranged in each shell, and two ends of each main channel are respectively communicated with the sample introduction channel and the sample outlet channel;
a plurality of connecting channels are arranged between any two adjacent shells, and two ends of each connecting channel respectively correspond to one capturing groove and are communicated with the upper part of the capturing groove.
2. The suspended array microfluidic chip of claim 1, wherein:
the sample outlet channel comprises at least one S-shaped curve.
3. The suspended array microfluidic chip of claim 2, wherein:
the height dimension of the connecting channel is not more than 10 micrometers, and the width dimension of the connecting channel is not more than 10 micrometers; and/or the length of the connecting channel is 5-200 microns.
4. The suspended array microfluidic chip of claim 3, wherein:
the height of the main channel and the height of the capture groove are both 30-60 micrometers.
5. The suspended array microfluidic chip of claim 4, wherein:
the projection length of the connecting part of the capturing groove and the main channel on the substrate layer is 20-40 micrometers; and/or the width of the main channel is 50-300 microns.
6. The suspended array microfluidic chip of claim 5, wherein:
the distance between the central lines of any two adjacent connecting channels is not less than the projection length.
7. The suspended array microfluidic chip of any one of claims 1 to 6, wherein:
the included angle between the connecting channel and the length direction of the main channel is 45-135 degrees.
8. The suspended array microfluidic chip of any one of claims 1 to 6, wherein:
the projection shape of the capture groove on the substrate layer is fan-shaped.
9. The suspended array microfluidic chip of any one of claims 1 to 6, wherein:
the suspended array microfluidic chip further comprises a supporting layer, and the channel layer is located between the supporting layer and the substrate layer.
10. The suspended array microfluidic chip of claim 9, wherein:
the supporting layer and the channel layer are made of any one of rigid plastics or glass.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113088447A (en) * | 2021-03-11 | 2021-07-09 | 温州医科大学 | Suspended array microfluidic chip and preparation method and application thereof |
| CN115044536A (en) * | 2022-06-20 | 2022-09-13 | 中国农业科学院农业质量标准与检测技术研究所 | Preparation method of 3D organ chip based on flow assembly |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113088447A (en) * | 2021-03-11 | 2021-07-09 | 温州医科大学 | Suspended array microfluidic chip and preparation method and application thereof |
| CN115044536A (en) * | 2022-06-20 | 2022-09-13 | 中国农业科学院农业质量标准与检测技术研究所 | Preparation method of 3D organ chip based on flow assembly |
| CN115044536B (en) * | 2022-06-20 | 2024-02-02 | 中国农业科学院农业质量标准与检测技术研究所 | Preparation method of 3D organ chip based on flow assembly |
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Correction item: Description|Drawings Correct: Delete the relevant explanation of Figure 14|Delete Figure 14 False: Contains relevant explanations for Figure 14|Containing Figure 14 Number: 50-01 Page: ?? Volume: 37 |
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| CU01 | Correction of utility model |