CN110058029B - Image acquisition and processing-free single cell cytoplasm viscosity measuring device and method - Google Patents
Image acquisition and processing-free single cell cytoplasm viscosity measuring device and method Download PDFInfo
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Abstract
The invention discloses a single cell cytoplasm viscosity measuring device and method without image acquisition and processing, wherein the device comprises: a microfluidic chip module; a pressure control module connected to the microfluidic chip module; the electricity generation and collection module is connected with the microfluid chip module; and the data analysis and processing module is connected with the electricity generation and acquisition module. The single cell cytoplasm viscosity measuring device provided by the invention obtains the cell cytoplasm viscosity through the time difference of two voltage changes generated when the single cell moves in the microfluidic chip module, so that the device can obtain related parameters without an image acquisition device, the device cost is reduced, and the image-free high-precision measurement of the single cell viscosity is realized.
Description
Technical Field
The invention relates to the technical field of microfluidics, in particular to a single cell cytoplasm viscosity measuring device and method without image acquisition and processing.
Background
Cells are the basic unit of all life activities, and in the life activities of a living body, cell division, differentiation and apoptosis continuously occur. As early as 1665, the uk scientist Robert Hooker first discovered cells, and since then, the human world has begun to continue exploring. In recent years, the study of single cells has attracted more and more researchers. It is widely accepted by the scientific community that cell heterogeneity exists not only among different types of cells but also among the same cells, and may be manifested by various biophysical and biochemical characteristics of the cells. Previous measurements of whole cell masses often have difficulty showing this difference because the measurement of whole cell masses often results as an average of all cell properties, and some significant differences between individual cells can be easily masked. In this context, single cell analysis techniques have emerged, which allow accurate measurement and characterization of individual cells, and which will have long-term implications for the diagnosis, treatment and human cell search of various diseases.
One important aspect of single cell analysis is the characterization of the various mechanical properties exhibited by individual cells. In the case of eukaryotic cells, the main components of the cell include cell membrane, cytoplasm, nucleus, etc., and the cytoplasm contains abundant microwires, microtubules, and intermediate filaments, which all form the cytoskeleton supporting the cell morphology and determine the response of the cell to various external forces. The vital movement of a cell is often accompanied by changes in the structures of cytoskeleton, cell membrane, cytoplasm, etc., and the corresponding changes in mechanical properties are also often shown. The cytomechanical properties often reflect changes in cell physiology. The cytoplasmic viscosity of a single cell is an important mechanical characteristic parameter of the single cell, and many studies indicate the relation between the cytoplasmic viscosity of the cell and diseases. For example, after a person becomes infected with malaria parasites, the cytoplasmic viscosity of erythrocytes, which are parasitic cells of malaria parasites, increases. In another example, the cytoplasmic viscosity of red blood cells is higher in sickle anemia patients compared to normal individuals, and the red blood cells often take on a sickle or crescent shape. Malignant cells multiply indefinitely in the body because of an abnormal mechanism of apoptosis and a smaller cytoplasmic viscosity.
In the aspect of characterizing the mechanical properties of single cells, the traditional methods include Atomic Force Microscopy (AFM) and micropipette technology. The atomic force microscope mainly comprises a photodiode, a cantilever beam and a microprobe, wherein in the actual measurement process, the microprobe is in contact with the surface of a cell and can cause the deformation of the cantilever beam, the photodiode at the rear end of the cantilever beam can sense the deformation and output the deformation, and finally the mechanical characteristic parameters of the cell are obtained. However, the method can only measure local mechanical properties of adherent cells, cannot measure the overall mechanical properties of the cells, and the detection result is often influenced by a cell substrate and a probe, and in addition, the detection flux is low, so that the data volume of the single cell mechanical properties reported by the method is very small at present. The traditional micro-suction tube technology utilizes negative pressure to attract cells to enter the opening part of a capillary tube, the cells form micro deformation at the opening part of the micro-suction tube and are then discharged, and the overall mechanical characteristic parameters of the cells can be calculated by analyzing the micro deformation of the cells in the micro-suction tube.
The characteristic dimension of the microfluidic technology is very close to the diameter of the cell, so that the single cell can be conveniently manipulated and analyzed, and the microfluidic technology has the characteristics of small sample demand and sensitive reflection. In recent years, methods for detecting mechanical properties of single cells based on microfluidic technology have attracted much attention, and typical methods include an electrical stretching method, a fluid stretching method, and a compressed channel method. Sun in 2011, an electrical stretching method is proposed, in which two plates with different sizes are used to generate an uneven electric field, cells move to a plate with a high electric field intensity in the uneven electric field, and when the cells reach the plate, the cells are static, and at the moment, the cells are subjected to electrostatic force to generate stretching deformation, and a corresponding mechanical model is combined, so that the young modulus of the mechanical property parameter of the cells can be obtained, but the method has low flux, and only results of less than 10 cells are reported. Prof.di Carlo proposed a fluid stretching method in 2012, in which a fluid shear force is used to deform cells, and the deformation of the cells is combined with a corresponding mechanical model, so as to obtain mechanical characteristic parameters of the cells, but the stress of the cells in a channel is different to a great extent according to different positions of the cells in a microfluid, so that the measurement result is not accurate. Chen proposed a method based on the mechanical properties of single cells of a compression channel in 2014, which utilizes negative pressure to attract cells to enter the compression channel, the cells deform in the process of entering the compression channel, and the deformation is combined with a mechanical model to realize the characterization of the young modulus of the single cells.
The current method for detecting the single cell mechanical property based on the microfluidic technology increases human understanding of the single cell, and the cytoplasmic viscosity of the cell is taken as an important single cell mechanical property parameter and is reported to be related to various diseases for many times, but due to the limitation of detection flux, a large amount of data of the cytoplasmic viscosity of the single cell still lacks. The measurement methods of the cytoplasm viscosity usually need to pass through the image acquisition and processing process, so the detection flux is seriously limited by a plurality of factors such as the image acquisition speed, the storage space, the image acquisition quality, the image data processing speed and the like, and a high-flux method of measuring the single cell cytoplasm viscosity without the image acquisition and processing process does not exist.
Therefore, there is an urgent need to develop an image-free high-throughput single-cell cytoplasmic viscosity measurement method and apparatus.
Disclosure of Invention
Technical problem to be solved
The present invention provides a single cell cytoplasm viscosity measuring device and method without image acquisition and processing, which at least partially solves the technical problems.
(II) technical scheme
According to an aspect of the present invention, there is provided an image-acquisition-and-processing-free single-cell cytoplasmic viscosity measurement apparatus, comprising:
the microfluid chip module is used for the movement of the single cell to be measured and the formation of the stretching-in;
the pressure control module is connected with the microfluidic chip module and is used for driving and controlling the single cell to move;
the electricity generation and collection module is connected with the microfluid chip module and is used for providing a voltage signal to the microfluid chip module, monitoring the voltage signal and recording the time of voltage change;
and the data analysis and processing module is connected with the electricity generation and acquisition module and is used for obtaining the extending speed of the single cell according to the time of voltage change, so that the viscosity of the single cell is obtained.
In further embodiments, the microfluidic chip module comprises:
a flow-through channel for movement of said single cell along said flow-through channel;
a movement measurement channel, which is crossed with the flow-through channel and is used for forming a part of the single cell in the movement measurement channel to extend into the movement measurement channel so as to generate a first voltage change;
a motion monitoring window located on the motion measurement channel for forming a partial protrusion of the single cell in the motion monitoring window for a second voltage change;
the first electrode and the second electrode are respectively positioned at two ends of the motion measurement channel and are used for collecting voltage signals;
and the third electrode and the fourth electrode are respectively positioned at two ends of the motion monitoring window and are used for acquiring voltage signals.
In further embodiments, the microfluidic chip module further comprises:
a cell inlet connected to the pressure control module;
a cell inflow channel connected to the cell inlet and one end of the flow channel;
a cell outflow channel connected to the other end of the flow channel;
a cell outlet connected to the cell outflow channel.
In a further embodiment, the pressure control module comprises:
a pressure source;
the pressure controller is connected with the pressure source and is used for outputting pressure and controlling the magnitude of the output pressure;
and the closed hose and the closed hard tube are connected with the pressure controller and the microfluidic chip module and are used for applying pressure to the microfluidic chip module.
According to another aspect of the present invention, there is provided a single-cell cytoplasmic viscosity measurement method without image acquisition and processing, including:
the pressure control module drives the single cell to be detected to move in the microfluidic chip module and form an extension, and the microfluidic chip module generates voltage change;
the electricity generation and collection module records the time of the micro-fluid chip module for generating two voltage changes;
and the data analysis and processing module obtains the extending speed of the single cell according to the time difference of the two voltage changes of the microfluid chip module recorded by the electricity generation and acquisition module, thereby obtaining the single cell viscosity.
In a further embodiment, before the single cell to be tested moves and forms the protrusion in the microfluidic chip module, the method further comprises:
filling a solution in the microfluidic chip module;
single cells to be measured are added to the microfluidic chip module and suspended in the solution.
In a further embodiment, the pressure control module driving the single cell to be tested to move and form an intrusion in the microfluidic chip module, and the voltage variation of the microfluidic chip module comprises:
the pressure control module drives the single cell to enter a flow channel of the microfluidic chip module and move continuously along the flow channel;
the single cell moves to a cell movement measuring channel and forms a part extending in the cell movement measuring channel to form first voltage change;
the single cell is extended into the motion monitoring window, forming a second voltage change.
In further embodiments, said recording the time at which the microfluidic chip module undergoes a voltage change comprises: and recording the time of the first voltage change and the second voltage change, and obtaining the time difference of the two voltage changes.
In further embodiments, said deriving the single cell viscosity from the time difference between the electrical occurrence and the voltage change of the microfluidic chip module recorded by the acquisition module comprises:
obtaining the extending speed of the single cell in the motion measurement channel according to the time difference;
and combining the extending speed with the single cell viscosity characteristic model to obtain the single cell viscosity.
In a further embodiment, the calculation formula of the penetration speed is:
wherein the content of the first and second substances,
the speed of the single cell extending into the movement measurement channel, L the distance from the movement monitoring window to the flow channel, and t1 and t2 the time of the first voltage change and the second voltage change, respectively.
(III) advantageous effects
According to the technical scheme, the single cell cytoplasm viscosity measuring device and the single cell cytoplasm viscosity measuring method which are free of image acquisition and processing have at least one of the following beneficial effects:
the single cell cytoplasm viscosity measuring device provided by the invention obtains the cell cytoplasm viscosity through the time difference of two voltage changes of the single cell when the single cell moves in the microfluidic chip module, so that the device does not need an image acquisition device to obtain cell deformation related parameters, the detection flux of the device is not limited by the acquisition speed of image acquisition equipment any more, the improvement of the detection flux is facilitated, and the device cost is obviously reduced.
The method for measuring the cytoplasm viscosity of the single cell can realize the measurement of the cytoplasm viscosity of the cell without using image acquisition equipment, namely, a complex image processing process is not needed, the detection process is simpler, and the calculation cost is reduced.
Drawings
FIG. 1 is a schematic structural diagram of a single-cell cytoplasmic viscosity measuring apparatus without image acquisition and processing according to the present invention;
FIG. 2 is a block diagram of a microfluidic chip module of the single-cell cytoplasmic viscosity measuring apparatus without image acquisition and processing according to the present invention;
FIG. 3 is a flow chart of the micro-fluidic chip of the single-cell cytoplasm viscosity measuring device without image collection and processing provided by the invention;
FIG. 4 is a schematic diagram of a data analysis and processing module 4 of the image-acquisition-and-processing-free single-cell cytoplasmic viscosity measurement apparatus provided in the present invention;
FIG. 5 is a flow chart of the method for measuring cytoplasmic viscosity of a single cell without image acquisition and processing according to the present invention.
Detailed Description
In order that the objects, technical solutions and advantages of the present invention will become more apparent, the present invention will be further described in detail with reference to the accompanying drawings in conjunction with the following specific embodiments.
According to the single cell cytoplasm viscosity measuring device and method provided by the invention, in the microfluidic chip module filled with the conductive medium solution, the single cell 106 deforms in the movement process, so that voltage change can be generated, the cell deformation speed is obtained through the time difference of two voltage changes caused by the movement of the single cell 106 in the microfluidic chip module, and the single cell cytoplasm viscosity can be obtained after the model is combined. The device does not need an image acquisition device to obtain related parameters, and can reduce equipment cost, improve flux and reduce calculation cost.
It should be noted that the claims and the description refer to the insertion of the single cell 106 generally as: during the movement of the single cell 106, part of the cell is squeezed into other channels that intersect the moving channel and is deformed.
As shown in fig. 1, fig. 1 is a schematic structural diagram of a single-cell cytoplasmic viscosity measurement device without image acquisition and processing, which includes:
the micro-fluid chip module 1 is used for moving in the single cell 106 to be measured and forming an extension;
the pressure control module 2 is connected with the microfluidic chip module 1 and is used for driving and controlling the single cell 106 to move;
the electricity generation and collection module 3 is connected with the microfluidic chip module 1 and is used for providing a voltage signal to the microfluidic chip module 1, monitoring the voltage signal and recording the time of voltage change;
and the data analysis and processing module 4 is connected with the electricity generation and acquisition module 3 and is used for obtaining the single cell viscosity according to the time of the voltage change. In this embodiment, the pressure control module 2 drives and controls the single cell 106 to be measured to move in the microfluidic chip module 1, the single cell 106 will have two or more voltage changes during its movement according to the design of the microfluidic chip module 1, the electricity generation and collection module 3 records the time difference between the two voltage changes therein, the data analysis and processing module 4 combines the time difference with the distance between the two voltage changes to obtain the insertion speed of the single cell 106, and the insertion speed is introduced into the single cell viscosity characteristic model to finally obtain the single cell viscosity.
The time of the voltage change is the time when the electricity generation and collection module 3 starts to change the voltage when monitoring the voltage signal of the microfluidic chip module 1.
Wherein, the calculation formula of the stretching speed is as follows:
wherein the content of the first and second substances,
the speed of the single cell 106 penetration, L, is the distance between two voltage changes, and t1 and t2 are the time of the first voltage change and the second voltage change, respectively.
As shown in fig. 2, fig. 2 is a structural diagram of a microfluidic chip module of the single-cell cytoplasmic viscosity measuring apparatus without image acquisition and processing provided by the present invention, where the microfluidic chip module 1 includes:
a flow-through channel 105 for movement of said single cell 106 along said flow-through channel 105;
a movement measurement channel 107 intersecting the flow channel 105 for forming a partial protrusion of the single cell 106 in the movement measurement channel 107 for a first voltage change;
a motion monitoring window 108 located on the motion measurement channel 107 for forming a partial protrusion of the single cell 106 in the motion monitoring window 108 for a second voltage change;
a first electrode 109 and a second electrode 110, which are respectively located at two ends of the motion measurement channel 107 and are used for collecting voltage signals;
and a third electrode 111 and a fourth electrode 112, which are respectively located at two ends of the motion monitoring window 108 and are used for collecting voltage signals.
The microfluidic chip module 1 comprises a cross-type compression channel, which in this embodiment is formed by the flow channel 105 and the motion measurement channel 107. Wherein, the flow channel 105 refers to a channel with one or two dimensions of any dimension of the cross section smaller than the diameter of the cell, and the diagonal dimension of the channel is generally 5-20 microns; the motion measurement channel 107 means that the length of one or two dimensions in any dimension of the cross section of the motion measurement channel is smaller than the diameter of the cell, the length of one or two dimensions in any dimension of the cross section of the motion measurement channel is smaller than the length of any dimension of the cross section of the main compression channel, and the diagonal dimension of the channel is 2-20 micrometers; the motion monitoring window 108 means that the length of one or two of the arbitrary dimensions of the cross section is smaller than the diameter of the cell, and the length of one or two of the arbitrary dimensions of the cross section is smaller than the length of the arbitrary dimension of the cross section of the main compression channel, and the diagonal dimension of the channel is 2-20 micrometers. In addition, the flow channel 105 and the motion measurement channel 107 may intersect perpendicularly or at any other angle, and the intersecting position may be located at any position of the flow channel 105; the motion monitoring window 108 may be located on one or both sides of the motion measurement channel 107; the cross-section of the motion measurement channel 107, the cell entrance channel, the cell exit channel 104 and the motion monitoring window 108 may be, but is not limited to, rectangular, circular or oval, and all of the above designs may perform the functions of the present invention.
In this embodiment, the single cell 106 moves along the flow channel 105, and when reaching the cell movement measuring channel 107, the single cell starts to form a partial protrusion into the movement measuring channel 107, at this time, the first electrode 109 and the second electrode 110 at two ends of the movement measuring channel 107 will detect a change of one voltage, as the cell continues to move along the cell flow channel 105, the protrusion length of the cell in the movement measuring channel 107 will gradually increase, and when it reaches the movement monitoring window 108, the third electrode 111 and the fourth electrode 112 at two ends of the movement monitoring window 108 will detect another voltageAfter which the cells leave the cell flow channel 105. Wherein one voltage change time detected by the first electrode 109 and the second electrode 110 is t1, the other voltage change time detected by the third electrode 111 and the fourth electrode 112 is t2, the distance from the motion monitoring window 108 to the flow channel 105 is the distance L forming two voltage changes, the penetration speed of the single cell 106 in the motion measurement channel 107 is the penetration speed of the single cell 106
As shown in fig. 2, the microfluidic chip module 1 may further include:
a cell inlet 101 connected to the pressure control module 2 for allowing the single cell 106 to enter the microfluidic chip module 1;
a cell inflow channel 103 connected to the cell inlet 101 and one end of the flow channel 105, for allowing the single cell 106 to smoothly enter the flow channel 105;
a cell outflow channel 104 connected to the other end of the flow channel 105 for flowing the single cell 106 out of the flow channel 105;
a cell outlet 102 connected to the cell outflow channel 104 for discharging the single cell 106 from the microfluidic chip module 1.
Wherein the cell inflow channel 103 and the cell outflow channel 104 refer to channels having a cross section larger than that of the cells, and a diagonal size of 30 to 1000 μm is generally used.
In this embodiment, the single cell 106 is added from the cell inlet 101, and then enters the cell inflow channel 103 and enters the flow channel 105 through the flow channel 105 under the driving of the pressure control module 2; after the single cell 106 is measured, it flows out of the flow channel 105 through the cell outflow channel 104, and finally exits the microfluidic chip module 1 through the cell outlet 102.
Further, the microfluidic chip module 1 further includes: a substrate (not shown) and a carrier closely combined therewith, on which the cell inlet 101, the cell inflow channel 103, the cell flow-through channel 105, the movement measurement channel 107, the movement monitoring window 108, the cell outflow channel 104 and the cell outlet 102 are formed. In this embodiment, the substrate is made of glass, and it should be clear to a person skilled in the art that, besides glass, the substrate may be made of a sheet material such as a silicon wafer, a poly methyl methacrylate (PMMA, Acrylic, or organic glass) or a poly dimethyl siloxane (PDMS) sheet. The material of the carrier in this embodiment is PDMS, and the carrier may be formed of glass, photoresist SU-8, silicon wafer, or other materials besides PDMS.
As shown in fig. 3, fig. 3 is a flow chart of the microfluidic chip fabrication of the single-cell cytoplasmic viscosity measurement apparatus without image acquisition and processing according to the present invention, and the fabrication steps include:
step a: spin-coating photoresist SU8-5 on a glass sheet, and performing alignment exposure for the first time;
step b: spin-coating a layer of photoresist SU8-25 on the photoresist SU8-5 of the step a, and performing alignment exposure for the second time;
step c: post-baking, developing and hardening to form a male mold of a cell inflow channel 103 and a cell outflow channel 104;
step d: c, pouring a mixed solution of PDMS and a curing agent on the mold obtained in the step c;
step e: demolding after curing to obtain the microfluidic channel;
step f: sputtering a layer of metal on a glass sheet;
step g: preparing an on-chip electrode with a certain shape on a glass sheet by a standard stripping process;
step h: and e, punching the microfluidic channel obtained in the step e and bonding the microfluidic channel with a glass sheet with an upper electrode.
In the present invention, the pressure control module 2 includes:
a pressure source for providing a pressure;
the pressure controller is connected with the pressure source and is used for outputting pressure and controlling the magnitude of the output pressure;
and the closed hose and the closed hard tube are connected with the pressure controller and the microfluidic chip module 1 and are used for applying pressure to the inside of the microfluidic chip module 1.
In this embodiment, the pressure source serves as a source for providing pressure; the pressure controller is used for outputting pressure and controlling the output pressure; the closed flexible pipe and the closed hard pipe are used for applying pressure to the microfluidic chip module. In general, the microfluidic chip module is filled with a solution, and the pressure control module 2 applies pressure to the solution to move the single cell 106 to be detected. The pressure control module 2 is prior art, and will not be described herein, and any other pressure control device capable of implementing the corresponding function may be used.
As shown in fig. 4, fig. 4 is a schematic diagram of a data analysis and processing module 4 of the single cell cytoplasmic viscosity measurement apparatus without image acquisition and processing provided by the present invention, where the data analysis and processing module 4 calculates an insertion speed of the single cell 106 according to a recorded distance between two voltage changes in combination with two voltage changes, and then inserts the insertion speed into the single cell viscosity characteristic model to obtain the single cell viscosity.
In the present embodiment, the single-cell viscosity characteristic model is a droplet model, i.e. the cells are regarded as the structure of water drops wrapped by a primary film, and the key parameters in the model are the applied pressure (Δ P), the equivalent radius (R) of the motion measurement channel 107p) The rate of change of depth of the cell in the motion measurement channel 107 with time
Cytoplasmic viscosity (. mu.) of cellsc) The relationship therebetween can be represented by the following formula:
by substituting the measured parameters into the above formulaThe cytoplasmic viscosity (. mu.) of the cells can be obtainedc). The final characterization parameter may be the cytoplasmic viscosity (. mu.) of the cellsc)。
As shown in fig. 5, fig. 5 is a flowchart of a single cell cytoplasmic viscosity measurement method without image acquisition and processing, which includes:
s1: the pressure control module 2 drives the single cell 106 to be detected to move in the microfluidic chip module 1 and form an extension, and the microfluidic chip module 1 generates voltage change;
s2: the electricity generation and collection module 3 records the time of the micro-fluid chip module 1 for generating two voltage changes;
s3: the data analysis and processing module 4 obtains the single cell viscosity according to the time difference of the two voltage changes of the microfluidic chip module 1 recorded by the electricity generation and acquisition module 3.
In the embodiment, the single cell cytoplasm viscosity can be obtained at low cost and high flux by monitoring and recording the time of two voltage changes, obtaining the extending speed of the single cell through the time difference, and obtaining the single cell viscosity through calculation according to the extending speed.
In this embodiment, before the single cell 106 to be tested moves and forms the protrusion in the microfluidic chip module 1, the method further includes:
s01: filling the microfluidic chip module 1 with a solution;
s02: the single cell 106 to be measured is added to the microfluidic chip module 1 and suspended on the solution.
In step S01, it is an experimental preparation stage, that is, the microfluidic device is first filled with liquid, and usually, a cell culture solution, Phosphate Buffered Solution (PBS) or physiological saline with the same osmotic pressure as the cells is used.
In step S1, the step of driving the single cell 106 to be tested to move and form an insertion in the microfluidic chip module 1 by the pressure control module 2, and the step of causing the microfluidic chip module 1 to generate the voltage change specifically includes:
s101: the pressure control module 2 drives the single cell 106 to enter the flow channel 105 of the microfluidic chip module 1 and move continuously along the flow channel 105;
s102: the single cell 106 moves to a cell movement measuring channel 107 and forms a part extending in the cell movement measuring channel to form a first voltage change;
s103: the single cell 106 extends into the motion monitoring window 108, creating a second voltage change.
In step S2, the recording the time when the voltage change occurs in the microfluidic chip module 1 includes: and recording the time of the first voltage change and the second voltage change, and obtaining the time difference of the two voltage changes.
In step S3, the obtaining the single-cell viscosity according to the time difference between the electrical generation and the voltage change of the microfluidic chip module 1 recorded by the acquisition module 3 comprises:
s301: obtaining the extending speed of the single cell 106 in the motion measurement channel 107 according to the time difference;
s302: and combining the extending speed with the single cell viscosity characteristic model to obtain the single cell viscosity.
The single cell viscosity characteristic model is the same as the model used in the detection device, and is not described herein again.
The present invention has been described in detail with reference to the accompanying drawings. From the above description, those skilled in the art should clearly understand the device and method for detecting the mechanical property of single cell 106 provided in the present invention.
It should be noted that the data analysis and processing module 4 may include various forms of computing devices, such as a general purpose computer, an Application Specific Integrated Circuit (ASIC), a Field Programmable Gate Array (FPGA), and the like. The data analysis and processing module 4 can operate according to various method flows as described above by loading programs, code segments, etc. stored in the storage device to implement functions such as image processing and cell cytoplasm viscosity calculation. The data analysis and processing module 4 may further include an input device, such as a mouse, a keyboard, etc., for inputting user commands, data, etc., and an output device, such as a display, etc., for outputting the processing results (e.g., prediction results, etc.) of the processor. The input device and the output device may be implemented in combination as a touch screen.
Unless otherwise indicated, the numerical parameters set forth in the specification and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by the present disclosure. In particular, all numbers expressing dimensions and so forth used in the specification and claims are to be understood as being modified in all instances by the term "about". Generally, the expression is meant to encompass variations of ± 10% in some embodiments, 5% in some embodiments, 1% in some embodiments, 0.5% in some embodiments by the specified amount.
In addition, unless steps are specifically described or must occur in sequence, the order of the steps is not limited to that listed above and may be changed or rearranged as desired by the desired design. The embodiments described above may be mixed and matched with each other or with other embodiments based on design and reliability considerations, i.e., technical features in different embodiments may be freely combined to form further embodiments.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are only exemplary embodiments of the present invention and are not intended to limit the present invention, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (9)
1. A single cell cytoplasm viscosity measuring device free of image acquisition and processing is characterized by comprising:
the micro-fluid chip module (1) is used for the movement of the single cell (106) to be measured and the formation of the extension; wherein the microfluidic chip module (1) comprises:
a flow-through channel (105) for movement of the single cell (106) along the flow-through channel (105);
a movement measurement channel (107) intersecting the flow-through channel (105) for forming a partial protrusion of the single cell (106) in the movement measurement channel (107) for a first voltage change;
a motion monitoring window (108) located on the motion measurement channel (107) for forming a partial protrusion of the single cell (106) in the motion monitoring window (108) for a second voltage change;
a first electrode (109) and a second electrode (110) which are respectively positioned at two ends of the motion measurement channel (107) and are used for collecting voltage signals;
the third electrode (111) and the fourth electrode (112) are respectively positioned at two ends of the motion monitoring window (108) and are used for collecting voltage signals;
the pressure control module (2) is connected with the microfluidic chip module (1) and is used for driving and controlling the single cell (106) to move;
the electricity generation and collection module (3) is connected with the microfluid chip module (1) and is used for providing a voltage signal for the microfluid chip module (1), monitoring the voltage signal and recording the time of voltage change;
and the data analysis and processing module (4) is connected with the electricity generation and acquisition module (3) and is used for obtaining the extending speed of the single cell (106) according to the time of the voltage change, so that the single cell viscosity is obtained.
2. The image acquisition and processing-free single-cell cytoplasmic viscosity measurement device according to claim 1, wherein said microfluidic chip module (1) further comprises:
a cell inlet (101) connected to the pressure control module (2);
a cell inflow channel (103) connected to the cell inlet (101) and one end of the flow channel (105);
a cell outflow channel (104) connected to the other end of the flow channel (105);
a cell outlet (102) connected to the cell outflow channel (104).
3. The image-acquisition-and-processing-free single-cell cytoplasmic viscosity measurement device according to claim 1 or 2, wherein the pressure control module (2) comprises:
a pressure source;
the pressure controller is connected with the pressure source and is used for outputting pressure and controlling the magnitude of the output pressure;
and the closed hose and the closed hard tube are connected with the pressure controller and the microfluidic chip module (1) and are used for applying pressure to the inside of the microfluidic chip module (1).
4. An image-acquisition-and-processing-free single-cell cytoplasmic viscosity measurement method using the image-acquisition-and-processing-free single-cell cytoplasmic viscosity measurement apparatus according to any one of claims 1 to 3, comprising:
the pressure control module (2) drives the single cell (106) to be detected to move in the microfluid chip module (1) and form an extension, and the microfluid chip module (1) generates voltage change;
the electricity generation and collection module (3) records the time of the micro-fluid chip module (1) for generating two voltage changes;
and the data analysis and processing module (4) obtains the extending speed of the single cell (106) according to the time difference of the two voltage changes of the microfluid chip module (1) recorded by the electricity generation and collection module (3), thereby obtaining the single cell viscosity.
5. The image-acquisition-and-processing-free method for measuring the cytoplasmic viscosity of a single cell as claimed in claim 4, wherein the step of moving and forming the protrusion of the single cell (106) to be measured in the microfluidic chip module (1) further comprises:
filling a solution in the microfluidic chip module (1);
adding a single cell (106) to be measured to the microfluidic chip module (1) and suspending the single cell (106) in the solution.
6. The image-acquisition-and-processing-free method for measuring the cytoplasmic viscosity of a single cell as claimed in claim 5, wherein the step of driving the single cell (106) to be measured to move and form the protrusion in the microfluidic chip module (1) by the pressure control module (2), and the step of changing the voltage of the microfluidic chip module (1) comprises:
the pressure control module (2) drives the single cell (106) to enter a flow channel (105) of the microfluidic chip module (1) and move continuously along the flow channel (105);
the single cell (106) moves to a cell movement measuring channel (107) and forms a part extending in the cell movement measuring channel to form a first voltage change;
the single cell (106) extends into a motion monitoring window (108) to form a second voltage change.
7. The image-acquisition-and-processing-free single-cell cytoplasmic viscometry method according to claim 6, wherein said recording of the time of occurrence of voltage changes of said microfluidic chip module (1) comprises: and recording the time of the first voltage change and the second voltage change, and obtaining the time difference of the two voltage changes.
8. The image-acquisition-and-processing-free single-cell cytoplasmic viscometry method according to claim 7, wherein the obtaining of the single-cell viscometry from the time difference of voltage changes of the microfluidic chip module (1) recorded by the electrical generation and acquisition module (3) comprises:
obtaining the extending speed of the single cell (106) in the motion measurement channel (107) according to the time difference;
and combining the extending speed with the single cell viscosity characteristic model to obtain the single cell viscosity.
9. The image-acquisition-and-processing-free method for measuring cytoplasmic viscosity of a single cell according to claim 8, wherein the calculation formula of the penetration speed is as follows:
wherein the content of the first and second substances,
the speed of the single cell (106) entering the movement measurement channel (107), L the distance from the movement monitoring window (108) to the flow channel (105), and t1 and t2 the time of the first voltage change and the second voltage change, respectively.
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