CN116139952A - Microfluidic device for chemical damage to living single cell subcellular microregions - Google Patents

Microfluidic device for chemical damage to living single cell subcellular microregions Download PDF

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CN116139952A
CN116139952A CN202310127500.0A CN202310127500A CN116139952A CN 116139952 A CN116139952 A CN 116139952A CN 202310127500 A CN202310127500 A CN 202310127500A CN 116139952 A CN116139952 A CN 116139952A
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microfluidic
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microfluidic probe
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林金明
张强
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Tsinghua University
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Abstract

The invention discloses a microfluidic device for chemically damaging a living single-cell microcell, which comprises a microfluidic probe system and a sample control platform, wherein the microfluidic probe system is arranged above the sample control platform, and microfluid generated by a microfluidic probe of the microfluidic probe system can act on a target subcellular region on a target single cell to realize in-situ subcellular chemical damage; the microfluidic probe system comprises a microfluidic probe and a flow injection pump, wherein microfluid in each channel of the microfluidic probe is driven and controlled by the flow injection pump and can respectively inject double-flow-phase microfluid consisting of a central-phase fluid and a sheath-flow-phase fluid into a cell sample; the sample control platform comprises a support component with a temperature control effect and a cell culture dish positioned above the support component, wherein the cell culture dish is used for placing a cell sample to be stimulated and a cell culture solution. The invention can limit substances in a micron-scale space in an open space to realize in-situ subcellular chemical stimulation.

Description

Microfluidic device for chemical damage to living single cell subcellular microregions
Technical Field
The application relates to the technical field of subcellular chemical stimulation, in particular to a microfluidic device for chemically damaging a living single-cell subcellular micro-region.
Background
In the biological field, the behavior research of single cells can reveal the property of the single cells as living body units, and the behavior of the single cells can be displayed under a certain stimulus, so that the development of the stimulus method is significant to the behavior research. Traditional research is based on the overall stimulation of cells, while deep cell behavior properties require subcellular precision stimulation to be triggered, which requires highly accurate stimulation methods. As an important class of stimuli, chemical injury can cause cells to exhibit injury resistance, repair, and regeneration behavior, and therefore it is critical to develop accurate methods of subcellular chemical injury.
In the prior art, the micro-fluidic technology is utilized to realize the regulation and control of chemical substance distribution, however, the chemical substance is difficult to focus due to the rapid diffusion of molecules on a micro scale, so that the stimulation precision of the existing micro-fluidic technology does not reach the level of micro-scale subcellular. In addition, most of the traditional methods are closed microfluidic, and samples need to be injected into a microstructure in a microfluidic chip, so that in-situ stimulation on original samples cannot be realized; the existing open microfluidic method is not capable of effectively realizing accurate control of micrometer-scale fluid, and cannot meet the requirement of subcellular stimulation.
Disclosure of Invention
Based on this, in order to solve at least one of the problems that the closed microfluidic method in the prior art needs a sample to be injected into a microstructure in a microfluidic chip and cannot realize in-situ stimulation of an original sample, and the open microfluidic method has not yet effectively realized accurate manipulation of a micrometer-scale fluid and cannot meet the requirement of subcellular stimulation, it is necessary to provide a microfluidic device for chemically damaging a living single-cell subcellular microcell. The microfluidic device for chemically damaging the subcellular microcell of the living body can realize subcellular chemical stimulation by limiting substances to a micrometer-scale space in an open space.
An embodiment of the present application provides a microfluidic device for chemically damaging a single cell subcellular microzone of a living body.
The microfluidic device for chemically damaging the living single-cell subcellular micro-region comprises a microfluidic probe system and a sample control platform, wherein the microfluidic probe system is arranged above the sample control platform, and microfluid generated by a microfluidic probe of the microfluidic probe system can act on a target subcellular region on a target single cell to realize subcellular chemical damage; the microfluidic probe system comprises a microfluidic probe and a flow injection pump, wherein the microfluidic probe is connected to the positioning mechanism, microfluid in each channel of the microfluidic probe is driven and controlled by the flow injection pump, and the microfluidic probe can respectively inject double-flow-phase microfluid consisting of a central-phase fluid and a sheath-flow-phase fluid into a cell sample; the sample control platform comprises a support component with a temperature control effect and a cell culture dish positioned above the support component, wherein a cell sample to be stimulated and a cell culture solution are placed in the cell culture dish, a detection mechanism is arranged below the support component, and the detection mechanism is used for observing the cell sample and the microfluidic probe in real time.
In some embodiments, the microfluidic probe system further comprises a syringe to which the microfluidics in each channel of the microfluidic probe is connected by a tubing, the syringe being connected to and controlled by the flow syringe pump.
In some of these embodiments, the tubing between each channel of the microfluidic probe and the syringe is polytetrafluoroethylene tubing.
In some of these embodiments, the cell sample in the cell culture dish is at 10 2 ~10 4 Planting at a density of one per square centimeter.
In some embodiments, the microfluidic probe comprises an intermediate layer channel and side channels, at least one side channel is respectively arranged on two opposite sides of the intermediate layer channel, the intermediate layer channel is used for injecting microfluid, and the side channels are used for extracting the microfluid.
In some of these embodiments, the intermediate layer channel comprises a first intermediate sub-channel, a second intermediate sub-channel, and a third intermediate sub-channel, the first intermediate sub-channel, the second intermediate sub-channel, and the third intermediate sub-channel being spaced apart; the first intermediate sub-channel is used for injecting a center phase fluid containing chemical injury substances; the second intermediate sub-channel is used for injecting sheath fluid phase fluid without chemical injury substances; the third intermediate sub-channel is used for injecting cell buffer.
In some embodiments, at least one second middle sub-channel is respectively arranged at two sides of the first middle sub-channel, and at least one third middle sub-channel is respectively arranged at the outer side of the second middle sub-channel away from the first middle sub-channel.
In some of these embodiments, the polyethylene glycol phase containing sodium dodecyl sulfate is the center phase fluid, the dextran phase containing no sodium dodecyl sulfate is the sheath fluid phase fluid,
and/or the cell buffer is Du's phosphate buffer;
and/or the chemically damaging substance is sodium dodecyl sulfate.
In some of these embodiments, the positioning mechanism is an XYZ positioner that is capable of X-axis, Y-axis, and Z-axis movement.
In some embodiments, the sample manipulation platform further comprises a detection mechanism, wherein the detection mechanism is arranged below the support assembly, and the detection mechanism is a microscope, and an objective lens of the microscope faces towards the support assembly.
According to the microfluidic device for chemically damaging the living single-cell subcellular micro-region, the two aqueous phases are controlled through the microfluidic, so that chemical molecules are limited in the open space micron-level region, and chemical stimulation on any subcellular micro-region is realized. The microfluidic device for chemically damaging the living single-cell subcellular micro-region can realize subcellular chemical stimulation by limiting substances to a micrometer-scale space in an open space, and has important significance for life science research.
Compared with the prior art, the invention has the following beneficial effects due to the adoption of the technical scheme:
(1) The invention utilizes the affinity and hydrophobicity properties of the double-water aqueous phase chemical substances to limit the molecular distribution, and well solves the problem that the chemical substances are difficult to focus due to rapid molecular diffusion.
(2) The invention utilizes the microfluidic probe and the flow injection pump of the multilayer composite channel, improves the fluid control precision, and successfully realizes the accurate control of the micrometer-scale fluid.
(3) The invention can be used for researching the response behavior of single cells after different substances stimulate different subcellular micro-regions, and revealing the regulation mechanism of 'pulling one and moving the whole body'.
(4) The microfluidic probe of the invention does not contact cells in the use process, but controls the fluid to act on the cells, thereby avoiding the cells from being mechanically damaged.
(5) The invention can locally damage single cells, thereby promoting the cells to show damage resistance, repair and regeneration behaviors and analyzing.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings that are required to be used in the description of the embodiments will be briefly described below. It is obvious that the drawings in the following description are only some embodiments of the present application, and that other drawings may be obtained from these drawings without inventive effort to a person skilled in the art.
For a more complete understanding of the present application and the advantages thereof, reference is now made to the following descriptions taken in conjunction with the accompanying drawings. Wherein like reference numerals refer to like parts throughout the following description.
FIG. 1 is a schematic diagram of the overall structure of a microfluidic device for chemically damaging a single cell subcellular microzone of a living body according to an embodiment of the invention;
FIG. 2 is a schematic side view of the bottom fluid distribution of a microfluidic probe of a microfluidic device for chemically damaging a single cell subcellular microzone of a living body according to an embodiment of the present invention;
FIG. 3 is a bottom view of reagent solution distribution near a cell sample of a microfluidic device for chemically damaging a living single cell subcellular microzone according to an embodiment of the present invention.
Description of the reference numerals
10. Microfluidic devices for chemically damaging living single cell subcellular microareas; 100. a microfluidic probe system; 101. a microfluidic probe; 1011. a first intermediate sub-channel; 1012. a second intermediate sub-channel; 1013. a third intermediate sub-channel; 1014. a side channel; 200. a sample manipulation platform; 201. a support assembly; 202. a cell culture dish; 203. a detection mechanism; 20. and (3) a cell sample.
Detailed Description
In order that the above objects, features and advantages of the invention will be readily understood, a more particular description of the invention will be rendered by reference to the appended drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. The present invention may be embodied in many other forms than described herein and similarly modified by those skilled in the art without departing from the spirit of the invention, whereby the invention is not limited to the specific embodiments disclosed below.
In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the device or element being referred to must have a specific orientation, be configured and operated in a specific orientation, and therefore should not be construed as limiting the present invention.
Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In the description of the present invention, the meaning of "plurality" means at least two, for example, two, three, etc., unless specifically defined otherwise.
In the present invention, unless explicitly specified and limited otherwise, the terms "mounted," "connected," "secured," and the like are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; either directly or indirectly, through intermediaries, or both, may be in communication with each other or in interaction with each other, unless expressly defined otherwise. The specific meaning of the above terms in the present invention can be understood by those of ordinary skill in the art according to the specific circumstances.
In the present invention, unless expressly stated or limited otherwise, a first feature "up" or "down" a second feature may be the first and second features in direct contact, or the first and second features in indirect contact via an intervening medium. Moreover, a first feature being "above," "over" and "on" a second feature may be a first feature being directly above or obliquely above the second feature, or simply indicating that the first feature is level higher than the second feature. The first feature being "under", "below" and "beneath" the second feature may be the first feature being directly under or obliquely below the second feature, or simply indicating that the first feature is less level than the second feature.
It will be understood that when an element is referred to as being "fixed" or "disposed" on another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "upper," "lower," "left," "right," and the like are used herein for illustrative purposes only and are not meant to be the only embodiment.
In the description of the present invention, the meaning of a number is one or more, the meaning of a number is two or more, and greater than, less than, exceeding, etc. are understood to exclude the present number, and the meaning of a number is understood to include the present number. The description of the first and second is for the purpose of distinguishing between technical features only and should not be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated or implicitly indicating the precedence of the technical features indicated.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
The embodiment of the application provides a microfluidic device 10 for chemically damaging a living single-cell subcellular micro-region, so as to solve at least one of the problems that in the prior art, a closed microfluidic needs a sample to be injected into a microstructure in a microfluidic chip, in-situ stimulation of an original sample cannot be realized, and an open microfluidic method cannot effectively realize accurate control of a micrometer-scale fluid and cannot meet the requirement of subcellular stimulation. The microfluidic device 10 for chemically damaging a single cell subcellular micro-region of a living body will be described with reference to the accompanying drawings.
Referring to fig. 1, an exemplary microfluidic device 10 for chemically damaging a living single-cell subcellular micro-region is shown in fig. 1, and fig. 1 is a schematic structural diagram of the microfluidic device 10 for chemically damaging a living single-cell subcellular micro-region according to an embodiment of the present application. The microfluidic device 10 for chemically damaging living single cell subcellular microareas of the present application is capable of achieving subcellular chemical stimulation in open spaces by confining substances to a micrometer-scale space.
In order to more clearly illustrate the structure of the microfluidic device 10 for chemically damaging a living single cell subcellular micro-region, the microfluidic device 10 for chemically damaging a living single cell subcellular micro-region will be described below with reference to the accompanying drawings.
Referring to fig. 1, fig. 1 is a schematic structural diagram of a microfluidic device 10 for chemical damage to a living single cell subcellular micro-region according to an embodiment of the present application.
An embodiment of the present application provides a microfluidic device 10 for chemically damaging a single cell subcellular microzone of a living body. A microfluidic device 10 for chemically damaging a living single cell subcellular microzone includes a microfluidic probe system 100 and a sample manipulation platform 200. The microfluidic probe system 100 is disposed above the sample manipulation platform 200, and the microfluidic generated by the microfluidic probes 101 of the microfluidic probe system 100 can act on a target subcellular region on a target single cell to achieve subcellular chemical damage.
In particular, the microfluidic probe system 100 comprises a microfluidic probe 101 and a flow injection pump. The microfluidic probe 101 is capable of injecting a double-flow-phase microfluidic composed of a center-phase fluid and a sheath-flow-phase fluid into a cell sample, respectively. The microfluidics in each channel of the microfluidic probe 101 (the microfluidics used comprises a cell buffer and two mutually immiscible aqueous solutions, one phase in which the target chemical is prone to be located is the central phase, the other phase is the sheath fluid phase), is controlled by the flow syringe pump drive.
The sample manipulation platform 200 includes a support assembly 201 with a temperature control effect and a cell culture dish 202 positioned above the support assembly 201. The cell culture dish 202 is used for placing a cell sample 20 to be stimulated and a cell culture solution for cell immersion. A detection mechanism 203 is arranged below the support assembly 201, and the detection mechanism 203 is used for observing the cell sample 20 and the microfluidic probe 101 in real time. In the invention, the culture dish is filled with a cell culture solution and a cell sample 20 to be researched, so as to ensure the activity of the sample. The support assembly 201 carries a temperature controller for creating a 37℃cell culture environment. A detection mechanism 203 is positioned below the support assembly 201 for viewing the target cells in real time. Under the observation of the detection mechanism 203, the target single cell can be moved to the target position by adjusting the support member 201, and the activity can be maintained for a long time.
In some of these embodiments, the microfluidic probe system 100 further comprises a positioning mechanism. The microfluidic probe 101 is connected to a positioning mechanism. The positioning mechanism is used for realizing accurate regulation and control of the three-dimensional position of the microfluidic probe 101, and is not shown in the drawings.
In some of these embodiments, the support assembly 201 is capable of movement in the X-axis direction and the Y-axis direction.
In some of these embodiments, the support assembly 201 is an XY stage that is capable of movement along the X-axis direction and the Y-axis direction. The XY stage moves along the X-axis direction and the Y-axis direction, and corresponds to the X-axis direction and the Y-axis direction movement of the positioning mechanism, and the XY stage moves in the vertical direction on the horizontal plane.
In some of these embodiments, the microfluidic probe system 100 further comprises a syringe. The microfluidics in each channel of the microfluidic probe 101 is connected by tubing to a syringe, which is connected to and controlled by a flow syringe pump.
In some of these embodiments, the tubing between each channel of the microfluidic probe 101 and the syringe is polytetrafluoroethylene tubing.
In some of these embodiments, the cell sample 20 in the cell culture dish 202 is at 10 2 ~10 4 Planting at a density of one per square centimeter.
In some of these embodiments, the microfluidic probe 101 comprises an intermediate layer channel and a side channel 1014. Opposite sides of the middle layer channel are each provided with at least one side channel 1014. The intermediate layer channel is used for microfluidic injection. The side channel 1014 is used for microfluidic extraction.
In some of these embodiments, the middle layer channel includes a first middle subchannel 1011, a second middle subchannel 1012, and a third middle subchannel 1013. The first intermediate subchannel 1011, the second intermediate subchannel 1012, and the third intermediate subchannel 1013 are spaced apart. The first intermediate sub-channel 1011 is used to inject a center phase fluid containing a chemically damaging substance. The second intermediate sub-channel 1012 is used to inject sheath fluid phase fluid that is free of chemically damaging substances; the third intermediate sub-channel 1013 is used for injecting cell buffer. In this embodiment, one phase of the two-phase solution of the first and second intermediate sub-channels 1011 and 1012 having a stronger affinity for the target chemical injury substance is used as the central phase, and the other phase is used as the sheath fluid phase, and the affinity difference can prevent the target substance from diffusing from the central phase to the outer sheath fluid phase across the phase interface. The central phase fluid can be compressed in the micron-scale open space, and the target chemically damaging species within it do not diffuse out of the central phase fluid to which it is attached, and thus are also confined to the micron-scale region. The selective damage to the region can be achieved by aligning the region to the subcellular microzone of the target single cell.
In some embodiments, referring to fig. 1 and 2, the number of first intermediate subchannels 1011 is one, the number of second intermediate subchannels 1012 is two, and the number of third intermediate subchannels 1013 is two.
In some embodiments, at least one second middle subchannel 1012 is disposed on each side of the first middle subchannel 1011, and at least one third middle subchannel 1013 is disposed on each side of the second middle subchannel 1012 away from the first middle subchannel 1011.
The microfluidic probe 101 is a multi-layer composite structure, a first middle subchannel 1011 and a second middle subchannel 1012 of the middle layer are respectively used for injecting cell buffer solution and two mutually-insoluble aqueous phase solutions, and two single channels at the edge are used for extracting fluid. The first intermediate sub-channel 1011 of the intermediate layer is used for injecting a central phase in which the target chemical is dissolved in advance, the second intermediate sub-channels 1012 on both sides thereof are used for injecting a sheath flow phase containing no target chemical, and the two third intermediate sub-channels 1013 on the outermost edges are used for injecting a cell buffer. The central phase containing the target chemical is protected and extruded by the sheath fluid phase without the target chemical, and the two phases are refocused by the cell buffer, so that the central phase and the target chemical inside the central phase are limited in the open space micron-level area. The subcellular regions covered by the microdomains will be chemically damaged while other cellular regions are not affected.
In some of these embodiments, a polyethylene glycol (PEG) phase containing Sodium Dodecyl Sulfate (SDS) is used as the central phase fluid, wherein the SDS-containing PEG phase is compressed into a micron-sized region, selectively destroying the subcellular structure of the region, and a dextran (Dex) phase free of Sodium Dodecyl Sulfate (SDS) is used as the sheath fluid phase fluid.
In some of these embodiments, the polyethylene glycol-dextran (PEG-Dex) forms a two aqueous phase system and the cell buffer is Duvet Phosphate Buffer (DPBS).
In some of these embodiments, the chemically damaging substance is Sodium Dodecyl Sulfate (SDS).
In some of these embodiments, the positioning mechanism is an XYZ positioner. The XYZ positioner is capable of X-axis, Y-axis, and Z-axis movement.
In some of these embodiments, the sample manipulation platform 200 further comprises a detection mechanism 203. A detection mechanism 203 is arranged below the support assembly 201, wherein the detection mechanism 203 is a microscope. The objective lens of the detection mechanism 203 is oriented towards the support assembly 201.
The microfluidic device 10 for chemically damaging a subcellular microzone of a living organism according to the present invention comprises the following steps:
1) Different channels of the microfluidic probe 101 are connected to corresponding syringes connected to the flow injection pump through polytetrafluoroethylene tubes, and corresponding solutions are respectively filled in different pipelines.
2) The microfluidic probe 101 is fixed to a positioning mechanism, and the positioning mechanism is adjusted to adjust the microfluidic probe 101 to a proper position under the observation of an objective lens of the detection mechanism 203.
3) The support assembly 201 is adjusted to move the target single cell to a target position directly below the outlet below the first middle sub-channel 1011 of the microfluidic probe 101, and the positioning mechanism is finely tuned to control the distance between the bottom end of the microfluidic probe 101 and the sample.
4) The flow syringe pump is activated to drive the fluid, and the central fluid containing the target chemical damaging substance is compressed within the open space micron-scale region, thereby damaging the subcellular structure of the region.
The microfluidic device 10 for chemically damaging the living single-cell subcellular micro-region forms a two-aqueous-phase system through polyethylene glycol-dextran (PEG-Dex), controls the two-aqueous-phase through micro-flow control, so that chemical molecules are limited in an open space micron-level region, and chemical stimulation on any subcellular micro-region is realized. The microfluidic device 10 for chemically damaging the subcellular microcell of the living body can limit substances in the open space to the micron-scale space to realize subcellular chemical stimulation, and has important significance for life science research.
Compared with the prior art, the invention has the following beneficial effects due to the adoption of the technical scheme:
(1) The invention utilizes the affinity and hydrophobicity properties of the double-water aqueous phase chemical substances to limit the molecular distribution, and well solves the problem that the chemical substances are difficult to focus due to rapid molecular diffusion.
(2) The invention utilizes the microfluidic probe 101 of the multilayer composite channel and the flow injection pump to improve the fluid control precision and successfully realize the accurate control of the micrometer-scale fluid.
(3) The invention can be used for researching the response behavior of single cells after different substances stimulate different subcellular micro-regions, and revealing the regulation mechanism of 'pulling one and moving the whole body'.
(4) The microfluidic probe 101 of the present invention does not contact the cells during use, but rather manipulates the fluid to act on the cells, thereby avoiding mechanical damage to the cells.
(5) The invention can locally damage single cells, thereby promoting the cells to show damage resistance, repair and regeneration behaviors and analyzing.
In the foregoing embodiments, the descriptions of the embodiments are emphasized, and for parts of one embodiment that are not described in detail, reference may be made to related descriptions of other embodiments.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The foregoing examples illustrate only a few embodiments of the invention and are described in detail herein without thereby limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.

Claims (10)

1. The microfluidic device for chemically damaging the living single-cell subcellular micro-region is characterized by comprising a microfluidic probe system and a sample control platform, wherein the microfluidic probe system is arranged above the sample control platform, and microfluid generated by a microfluidic probe of the microfluidic probe system can act on a target subcellular region on a target single cell to realize subcellular chemical damage; the microfluidic probe system comprises a microfluidic probe and a flow injection pump, wherein microfluid in each channel of the microfluidic probe is driven and controlled by the flow injection pump, and the microfluidic probe can respectively inject double-flow-phase microfluid consisting of a center-phase fluid and a sheath-flow-phase fluid into a cell sample; the sample control platform comprises a support component with a temperature control effect and a cell culture dish positioned above the support component, wherein a cell sample to be stimulated and a cell culture solution are placed in the cell culture dish, a detection mechanism is arranged below the support component, and the detection mechanism is used for observing the cell sample and the microfluidic probe in real time.
2. The microfluidic device for chemical damage to a living single cell subcellular microzone according to claim 1, wherein the microfluidic probe system further comprises a syringe to which the microfluidics in each channel of the microfluidic probe is connected by tubing, the syringe being connected to and controlled by the flow syringe pump.
3. The microfluidic device for chemically damaging a subcellular microzone of a living organism according to claim 2, wherein the tubing between each channel of the microfluidic probe and the syringe is polytetrafluoroethylene tubing.
4. The microfluidic device for chemical damage to living single cell subcellular microareas of claim 1, wherein the cell sample in the cell culture dish is at 10 2 ~10 4 Planting at a density of one per square centimeter.
5. The microfluidic device for chemical damage to a living single-cell subcellular micro-region according to any one of claims 1-4, wherein the microfluidic probe comprises an intermediate layer channel and side channels, at least one of the side channels being provided on opposite sides of the intermediate layer channel, respectively, the intermediate layer channel being for microfluidic injection and the side channels being for microfluidic withdrawal.
6. The microfluidic device for chemically damaging a single-cell subcellular microzone of claim 5, wherein the intermediate layer channel comprises a first intermediate sub-channel, a second intermediate sub-channel, and a third intermediate sub-channel, the first intermediate sub-channel, the second intermediate sub-channel, and the third intermediate sub-channel being spaced apart; the first intermediate sub-channel is used for injecting a center phase fluid containing chemical injury substances; the second intermediate sub-channel is used for injecting sheath fluid phase fluid without chemical injury substances; the third intermediate sub-channel is used for injecting cell buffer.
7. The microfluidic device for chemical injury to a living single-cell subcellular micro-region according to claim 6, wherein at least one second intermediate sub-channel is disposed on each side of the first intermediate sub-channel, and at least one third intermediate sub-channel is disposed on each side of the second intermediate sub-channel away from the first intermediate sub-channel.
8. The microfluidic device for chemically damaging a single-cell subcellular micro-region of living body according to claim 6, wherein polyethylene glycol phase containing sodium dodecyl sulfate is used as the center phase fluid and dextran phase containing no sodium dodecyl sulfate is used as the sheath fluid phase fluid;
and/or the cell buffer is Du's phosphate buffer;
and/or the chemically damaging substance is sodium dodecyl sulfate.
9. The microfluidic device for chemical injury to a living single-cell subcellular micro-region according to any one of claims 1-4, 6-8, wherein the microfluidic probe system comprises a positioning mechanism, the microfluidic probe is connected to the positioning mechanism, and the positioning mechanism is used for realizing precise regulation of the three-dimensional position of the microfluidic probe; the support component can move along the X-axis direction and the Y-axis direction;
and/or the positioning mechanism is an XYZ positioner, and the XYZ positioner can move in the X-axis direction, the Y-axis direction and the Z-axis direction.
10. The microfluidic device for chemical damage to a living single cell subcellular micro-region according to any one of claims 1-4, 6-8, wherein the sample manipulation platform further comprises a detection mechanism, the detection mechanism is arranged below the support assembly, wherein the detection mechanism is a microscope, and an objective lens of the microscope faces towards the support assembly.
CN202310127500.0A 2023-02-17 2023-02-17 Microfluidic device for chemical damage to living single cell subcellular microregions Pending CN116139952A (en)

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