CN104946523B - The measurement apparatus and measuring method of a kind of cyto-mechanics - Google Patents
The measurement apparatus and measuring method of a kind of cyto-mechanics Download PDFInfo
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- CN104946523B CN104946523B CN201510282272.XA CN201510282272A CN104946523B CN 104946523 B CN104946523 B CN 104946523B CN 201510282272 A CN201510282272 A CN 201510282272A CN 104946523 B CN104946523 B CN 104946523B
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- G01N33/48707—Physical analysis of biological material of liquid biological material by electrical means
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Abstract
The present invention relates to a kind of measurement apparatus, the measurement apparatus and measuring method of a kind of cyto-mechanics are disclosed, the measurement apparatus includes：Sample disc (1), for placing cell sample；Measuring unit (2), for being contacted with the cell sample in the sample disc (1), obtains the mechanics parameter of the cell sample；And control unit (3), it is connected with the measuring unit (2), for controlling the measuring unit (2) to move vertically according to the mechanics parameter, to change the exposure level of the measuring unit (2) and the cell sample, the measuring unit (2) is set to obtain the mechanics parameter of the cell sample in during the exposure level changes.The measurement apparatus of cyto-mechanics of the present invention can accurately obtain the mechanics parameter of cell.
Description
Technical field
The present invention relates to a kind of measurement apparatus, in particular it relates to the measurement apparatus and measuring method of a kind of cyto-mechanics.
Background technology
The mechanics parameter of cell includes rigidity, viscosity of cell etc., and the state of the mechanics parameter and cytoskeleton is tight
Close correlation, and cytoskeleton has played crucial work during cell migration, propagation, phagocytosis and correlation molecule signal transduction
With.Pathophysiological process in each organ-tissue of human body is along with the change of cyto-mechanics parameter, particularly tumour cell, due to
The abilities such as its propagation, migration and metabolism are different from normal cell, and its mechanics parameter is significantly different from normal physiological group
The cell knitted.For example, the rigidity for coming from normal bladder tissue cell line (Hu609 and HCV29) comes from Bladder Cancer
Ten times of cell line (Hu456, T24, BC3726) rigidity；Come from normal galactophore tissue's cell line (MCF-10A) rigidity
Come from 1.4~1.8 times of breast cancer tissue's cell line (MCF-7) rigidity.In addition, the mechanics parameter of tumour cell can be with
React its sensitiveness to chemotherapeutics.Research finds that green tea crude extract (Green Tea Extract, GTE) is optionally
Cytotoxicity is produced to tumour cell, while not influenceing normal cell, after being acted on through GTE, tumour cell is due to actin table
Up to increase, stiffness by itself also increases；And normal cell acted on through GTE after rigidity without significant change.
Therefore, the detection of tumour cell itself mechanics parameter can be applied not only to the early diagnosis of cancer in clinic with
And the selection of personalized chemotherapeutic medicine, while or the quantitative foundation of high-throughout chemotherapeutics screening offer.
Atomic force microscopy is the common technology means for measuring cell rigidity, but traditional AFM can only be
Detecting to living cells in liquid phase physiological environment, obtains cell image, and technical parameter requirement is strict；Due to piezoelectric ceramic tube
Formula scanner resonant frequency is low, causes that measurement efficiency is low, accuracy of detection is poor, limits it in high-flux cell mechanical test
Further apply.
The content of the invention
It is an object of the invention to provide a kind of cyto-mechanics measurement apparatus, the mechanics parameter of cell can be accurately obtained.
To achieve these goals, the present invention provides a kind of measurement apparatus of cyto-mechanics, and the measurement apparatus includes sample
Product disk, for placing cell sample；Measuring unit, for being contacted with the cell sample in the sample disc, obtains the cell
The mechanics parameter of sample；And control unit, it is connected with the measuring unit, for being surveyed according to mechanics parameter control is described
Unit vertical movement is measured, to change the exposure level of the measuring unit and the cell sample, obtains the measuring unit
The mechanics parameter of the cell sample in during the exposure level change.
The measurement apparatus of cyto-mechanics of the present invention can adjust measuring unit and sample as needed by the setting of control unit
Exposure level in product disk between cell sample, with the mechanics parameter of inner cell sample during obtaining exposure level change, measurement
Scope is wide, the degree of accuracy is high.
Another object of the present invention is to provide a kind of measuring method of cyto-mechanics, the mechanics ginseng of cell can be accurately obtained
Number.
To reach above-mentioned purpose, the technical proposal of the invention is realized in this way：
A kind of measuring method of cyto-mechanics, the measuring method includes：Cell sample is positioned in sample disc；Use
Measuring unit measurement is contacted with the cell sample in the sample disc, obtains the mechanics parameter of the cell sample；According to described
Mechanics parameter controls the measuring unit vertical movement, to change the exposure level of the measuring unit and the cell sample,
The measuring unit is set to obtain the mechanics parameter of the cell sample in during the exposure level changes.
What the measurement apparatus of the measuring method of the cyto-mechanics and above-mentioned cyto-mechanics had relative to prior art
Dominant Facies are same, will not be repeated here.
Other features and advantages of the present invention will be described in detail in subsequent embodiment part.
Brief description of the drawings
Accompanying drawing is, for providing a further understanding of the present invention, and to constitute a part for specification, with following tool
Body embodiment is used to explain the present invention together, but is not construed as limiting the invention.In the accompanying drawings：
Fig. 1 is the measurement apparatus stereochemical structure block diagram of cyto-mechanics of the present invention；
Fig. 2 is the modular structure schematic diagram of the control unit in the measurement apparatus of cyto-mechanics of the present invention；
Fig. 3 be cyto-mechanics of the present invention measurement apparatus in sample disc structural representation；
Fig. 4 is the flow chart of the measuring method of cyto-mechanics of the present invention；
Fig. 5 be cyto-mechanics of the present invention measuring method in positioning table closed-loop control schematic diagram；
Fig. 6 is an embodiment of the measuring method of cyto-mechanics of the present invention.
Description of reference numerals
The measuring unit of 1 sample disc 2
The photoelectric conversion module of 21 laser 22
The piezoelectric ceramic stack of 23 probe detector 24
The control unit of 25 image capture module 3
The control module of 31 adjustment of displacement module 32
The positioning table control module of 33 photoelectric processing module 34
The positioning table of 35 displacement platform control module 4
5 displacement platforms
Embodiment
The embodiment of the present invention is described in detail below in conjunction with accompanying drawing.It should be appreciated that this place is retouched
The embodiment stated is merely to illustrate and explain the present invention, and is not intended to limit the invention.
In the present invention, in the case where not making opposite explanation, the direction term mentioned, such as " on ", " under ", " preceding ",
" rear ", "left", "right" etc., are only the directions of refer to the attached drawing.Therefore, the direction term used is intended to be illustrative and not intended to limit
Protection scope of the present invention.
As depicted in figs. 1 and 2, cyto-mechanics measurement apparatus of the present invention includes sample disc 1, for placing cell sample；Survey
Unit 2 is measured, for being contacted with the cell sample in the sample disc 1, the mechanics parameter of the cell sample is obtained；And control
Unit 3, is connected with the measuring unit 2, for controlling the measuring unit 2 to move vertically according to the mechanics parameter, to change
Become the exposure level of the measuring unit 2 and the cell sample, the measuring unit 2 is obtained the exposure level change phase
The mechanics parameter of the interior cell sample.
The measurement apparatus of cyto-mechanics of the present invention can adjust measuring unit and sample as needed by the setting of control unit
Exposure level in product disk between cell sample, with the mechanics parameter of inner cell sample during obtaining exposure level change, measurement
Scope is wide, the degree of accuracy is high；And to measuring environment without particular/special requirement, measurement efficiency is high.
Described control unit 3 controls the vertical movement of measuring unit 2 to include according to the mechanics parameter：The control is single
Mechanics parameter acquired in the measuring unit 2 is compared by member 3 with setting value, is used for：In the mechanics parameter>Setting value
When, the upward vertical movement of measuring unit 2 is controlled, weakens the measuring unit 2 and the exposure level of the cell sample,
The stop motion when mechanics parameter=setting value obtained again；In the mechanics parameter<During setting value, control is described
The vertical downward movement of measuring unit 2, strengthens the measuring unit 2 and the exposure level of the cell sample, until obtaining again
The stop motion during mechanics parameter taken=setting value.
Wherein, the mechanics parameter includes the interaction between cell rigidity, viscosity and measuring unit and cell sample
At least one of power etc..
As shown in figure 1, the measuring unit 2 includes laser 21, for launching light beam perpendicular to the sample disc 1；Inspection
Module is surveyed, can be moved vertically, for contacting and interacting with the cell sample in the sample disc 1, while swashing described
The light beam that light device 21 is launched is reflected into the spot signal for characterizing mechanics parameter；And photoelectric conversion module 22, it is described for receiving
The electric signal that spot signal and being converted to characterizes the mechanics parameter is sent to described control unit 3；Described control unit 3 is distinguished
The photoelectric conversion module 22 and detection module are connected, the electric signal of mechanics parameter is characterized for receiving, and according to the mechanics
Detection module described in state modulator moves vertically, to change the exposure level of the detection module and the cell sample.At this
In embodiment, the photoelectric conversion module 22 is four-quadrant photoelectric sensor.
Wherein, the detection module includes probe detector 23, for contacting and interacting with cell sample, simultaneously will
Light beam is reflected into the spot signal for characterizing mechanics parameter；And piezoelectric ceramic stack 24, it is connected with the probe detector 23；For
Controlled by described control unit 3, and drive the probe detector 23 to move vertically.
In the present embodiment, the probe detector 23 includes probe, and the probe is installed on probe base, the probe
It is rigidly connected with the piezoelectric ceramic stack 24.
So that the mechanics parameter is interaction force as an example：The interaction that described control unit 3 obtains the probe
Power is compared with setting value, when the interaction force>During setting value, described control unit 3 controls the piezoelectric ceramic stack
24, in upward vertical movement, weaken the exposure level of the probe and cell sample, until the interaction force obtained again
Stop during=setting value；Work as interaction force<During setting value, control unit 3 controls the piezoelectric ceramic stack 24 vertically downward
Motion, making the exposure level of the probe and cell sample strengthens, and stops when the interaction force=setting value obtained again
Only.The piezoelectric ceramic stack 24 is by the way that motion makes probe complete to a cell sample surface up and down in vertical direction
Measurement, obtain multigroup mechanics parameter in during exposure level changes, form mechanics parameter curve, and single by the control
First 3 record storage related datas.
The measuring unit 2 also includes fine setting module (not shown), is connected, is used for the photoelectric conversion module 22
The position of the photoelectric conversion module 22 is adjusted to receive the spot signal.
Preferred scheme, the measuring unit 2 also includes image capture module 25, and the detection is shown for gathering in real time
The image of the position relationship of module and cell sample, to ensure that the detection module is vertical with the cell sample corresponding.At this
In embodiment, described image acquisition module 25 is CCD (Charge-coupled Device, charge coupled cell).Pass through CCD
Setting, the position relationship of the probe and cell sample can be obtained in real time, so as in the position of the probe and cell sample
Occur to be adjusted in time during deviation, it is ensured that the accuracy of mechanics parameter measurement.
Such as Fig. 2, described control unit 3 includes adjustment of displacement module 31, for controlling the detection module to move vertically；With
And control module 32, the photoelectric conversion module 22 and the adjustment of displacement module 31 are connected respectively, it is described for receiving sign
The electric signal of mechanics parameter determines corresponding mechanics parameter, and exports vertical displacement signal to institute's rheme according to the mechanics parameter
Move adjusting module 31.
The electric signal of mechanics parameter is characterized to make the control module 32 accurately receive, in addition it is also necessary to which the electric signal is entered one
Step processing, i.e.,：Described control unit 3 also includes photoelectric processing module 33, is arranged at the control module 32 and opto-electronic conversion mould
Between block 22, for that will be inputted after the electric signal enhanced processing of the photoelectric conversion module 22 to the control module 32.
As shown in figure 1, the measurement apparatus of cyto-mechanics of the present invention also includes：Positioning table 4, is connected with described control unit 3,
And the sample disc 1 is provided with the positioning table 4, for being controlled by described control unit 3, and drive the water of sample disc 1
Flat and/or vertical movement, so that the measuring unit 2 measures the different cell samples of same measured zone in the sample disc 1.
Corresponding, described control unit 3 also includes positioning table control module 34 (as shown in Figure 2), and the control is connected respectively
Molding block 32 and positioning table 4, are used for：Controlled by the control module 32, control the level of positioning table 4 and/or vertical movement；
Or the horizontal displacement signal of the positioning table is fed back into the control module 32, the control module 32 is according to the positioning
The horizontal displacement signal of platform 4 controls the positioning table 4 to continue to move horizontally or stop.
Wherein, the method that the control positioning table is continued to move to or stopped includes：Calculate the level of the positioning table
Whether the difference of displacement and setting displacement, judge the difference in setting difference range；If in setting difference range,
The positioning table is controlled to stop movement；Otherwise, the positioning table is controlled to continue to move horizontally.
In the pointwise position fixing process of positioning table 4, in order that the positioning table 4 is stopped as early as possible after reaching target location,
The PID closed-loop controls (as shown in Figure 5) in dead band are can adjust to employing band in the control of the positioning table 4 in the present invention.Root
The need for according to use, it can set when positioning table goes to ± n nanometers of distance objective position, positioning table control module 34 stops watching
Clothes control, makes positioning table 4 keep current location motionless.
When the control signal r (k) that positioning table control module is exported passes through conversion as shown in Figure 5, one-time positioning control is completed
After system, located control module of displacement signal c (k) is fed back to by displacement transducer by control module.The control module
R (k) and c (k) is subjected to subtraction process, difference e (k) is obtained, and determine whether the e (k) whether in setting difference range
[-e0,e0] in：When setting difference range in (i.e. | e (k) |≤e0) when, stop SERVO CONTROL, i.e., described positioning table stops fortune
It is dynamic, proceed by the measurement of mechanics parameter；Conversely, ought not setting difference range in (i.e. | e (k) |>e0) when, then continue servo
Control, i.e., described positioning table continues horizontal movement, until | e (k) |≤e0。
By above-mentioned closed-loop control, positioning time is not only shortened, and during avoiding mechanical meaurement, due to above-mentioned
Sample is moved caused by positioning table servo action, improves mechanical meaurement precision.
In addition, positioning table control module 34 can also control positioning table 4 to carry out the motion of vertical direction (i.e. Z-direction), use
Caused measurement error is tilted in compensation cell sample or measuring unit 2.
In the present embodiment, the positioning table 4 is flexible hinge nanometer positioning platform.Wherein, the flexible hinge nanometer is fixed
Position platform can realize the quick positioning of the small range nano level high resolution power in horizontal direction (i.e. X, Y-direction), such as the μ of 200 μ m 200
M, Z-direction stroke is several μm, and such as 4 μm, resolving power reaches sub- nanometer scale, and the flexible hinge nanometer positioning stylobate is in piezoelectric ceramics
Heap driving, mechanical lever amplification principle, high-pressure activated is exported by control unit.
Further, the measurement apparatus of cyto-mechanics of the present invention also includes displacement platform 5, is connected with described control unit 3, and
The positioning table 4 is provided with the displacement platform 5, for being controlled by described control unit 3, drive the level of positioning table 4 and/
Or vertical movement, so that the measuring unit 2 measures the cell sample of measured zones different in the sample disc.
Corresponding, described control unit 3 also includes displacement platform control module 35 (as shown in Figure 2), and the control is connected respectively
Molding block 32 and displacement platform 5, are used for：Controlled by the control module 32, control the level of displacement platform 5 and/or vertical movement；
Or the horizontal displacement signal of the displacement platform 5 is fed back into the control module 32, the control module 32 is according to institute's rheme
The horizontal displacement signal of moving stage 5 controls the displacement platform 5 to continue to move horizontally or stop.
Wherein, the control module 32 controls the displacement platform 5 to continue water according to the horizontal displacement signal of the displacement platform 5
The method that translation is moved or stopped is similar with the control method of positioning table 4 described above, will not be repeated here.
The displacement platform 5 can drive positioning table 4 thereon to carry out X, move in Y-direction, with handover measurement region, move model
Enclose for 1 × 1cm2-50×50cm2, and Z-direction motion is carried out, to realize that sample disc is forced probe behind handover measurement region again
Near and far is moved from the Z-direction of action, and range of movement is 1-50cm.
Described control unit 3 in the measurement apparatus of cyto-mechanics of the present invention can also connect host computer, to realize instruction
Control, the collection and output of data.Wherein, the control module 32 can be PC104 embedded main boards, and the PC104 is embedded
Mainboard, by network connection, the adjustment of displacement module 31, photoelectric processing is connected by PC104 buses respectively with the host computer
Module 33, positioning table control module 34 and displacement platform control module 35.
As shown in figure 3, the sample disc 1 includes multiple measured zones, each measured zone is into the chessboard of dot matrix
Lattice distribution.The measurement range of each measured zone is 1 × 1mm2To 30 × 30mm2, each tessellated region for 10 ×
10μm2To 50 × 50 μm2；It is adjacent tessellated at intervals of 0-10 μm.Further, have inside each gridiron pattern and promote carefully
The adherent promotion layer of born of the same parents, the promotion layer can be by collagen, laminin, fibronectin, gelatin and poly-D-lysine
Material Deng any one or combination is made, or other protein materials for having correlation properties are made；Adjacent chessboard compartment tool
There is the inhibition layer for suppressing cell attachment, the inhibition layer can be by such as material system such as bovine serum albumin(BSA), polyethylene glycol and F127
Into.The sample disc 1 can be fastened on positioning table 4 by screw, vacuum suction or magnetic-adsorption.
The measurement apparatus of cyto-mechanics of the present invention has the advantages that locating speed is fast, precision is high using positioning table, carries out X, Y
The quick and resetting of direction a small range, makes rapidly the central position of the cell sample to be measured in detection module alignment sample disc
Put；Have the advantages that moving range is big, precision is high, load capacity is strong using displacement platform, make its positioning table and sample of drive thereon
Product disk automatic switchover sample measured zone, it is to avoid because positioning table moving range is small, the manual switching measured zone disadvantage that time-consuming
End；Using the advantage that piezoelectric ceramic stack resonant frequency is high, positioning precision is high, the survey of high-precision mechanics parameter can be quickly carried out
Examination；Make cell sample ordered arrangement using sample disc gridiron pattern alignment technology, make the process of the cell measurement in whole sample disc,
First cell sample and probe alignment only need to be searched out by CCD and displacement platform, then by simply controlling positioning table
It can be achieved with quick and precisely positioning remaining cell with displacement platform, easy to operate, measurement efficiency is high.
As shown in figure 4, the measuring method of cyto-mechanics of the present invention includes：Step 401：Cell sample is positioned over sample disc
In；Step 402：Measuring unit is contacted with the cell sample in the sample disc, obtain the mechanics parameter of the cell sample；
Step 403：The measuring unit vertical movement is controlled according to the mechanics parameter, to change the measuring unit and the cell
The exposure level of sample, makes the measuring unit obtain the mechanics ginseng of the cell sample in during the exposure level changes
Number.
Wherein, the method for controlling the measuring unit to move vertically according to the mechanics parameter includes：By the power
Parameter is learned to be compared with setting value：In the mechanics parameter>During setting value, the measuring unit upward vertical movement is controlled,
Weaken the measuring unit and the exposure level of the cell sample, until the mechanics parameter=setting value obtained again
When stop motion；In the mechanics parameter<During setting value, the measuring unit vertical downward movement is controlled, makes the measurement single
First exposure level with the cell sample strengthens, the stop motion when mechanics parameter=setting value obtained again.
The mechanics parameter includes in interaction force between cell rigidity, viscosity and measuring unit and cell sample etc. at least one
Person.
Wherein, the measuring method of cyto-mechanics of the present invention also includes：Before step 401 operation, sample disc should be first prepared,
I.e.：A piece of template is provided, micron post is prepared in the template, makes chessboard point of the micron post into dot matrix in silicon template
Cloth, forms the sample disc, wherein, one measured zone of multiple gridiron pattern formation, the sample disc includes multiple measured zones.
Further, the sample disc is modified with rush in the lower processing that imposes a condition, the upper surface for making the micron post
Enter the promotion layer of cell attachment, adjacent micron intercolumniation has the inhibition layer for suppressing cell attachment.
Wherein, preparing the method for micron post includes：Silicon template is prepared using coupled plasma etch technology is reacted, according to
Template carries out complex using dimethyl silicone polymer (PDMS, polydimethylsiloxane), obtains the structure of micron post.Its
In, the rectangular parallelepiped structure that the micron post is cylinder or side is square, the diameter or size dimension of micron post are 10-50 μ
M, is highly 5-10 μm, and the spacing of adjacent microtrabeculae is 0-10 μm, multiple microns of post one measured zone of formation, the measured zone
Area be 1 × 1mm2-30×30mm2。
It is described to include in the processing that imposes a condition：The micron rod structure is subjected to corona treatment, 10- is then soaked in
100 μ g/mL collagen, laminin, fibronectin, gelatin, or in 0.01% dextrorotation Poly-L-Lysine Solution,
37 DEG C are incubated 1 hour, to have modified the micron post of mentioned component as seal, by western blot in hydrophobic plastic ware or glass dish
The promotion layer for promoting cell attachment is modified with the surface of substrate, the upper surface for making the micron post；And it is modified with rush by above-mentioned
The plastic ware or glass dish substrate for entering the promotion layer of cell attachment are incubated in F-127, bovine serum albumin(BSA) or polyglycol solution
Educate after half an hour, make that there is the inhibition layer for suppressing cell attachment between adjacent chessboard.Cell sample is put into above-mentioned treated sample
The measurement of mechanics parameter is carried out in product disk, and after cultivating 0.5-12 hours.
The measuring method of cyto-mechanics of the present invention also includes：The sample disc is installed on positioning table, it is described fixed to control
Position and/or is vertically moved platform level, the detection unit is directed at cell sample to be measured；Terminate in a cell sample measurement
Afterwards, the positioning table level and/or vertical movement are controlled, the measuring unit is measured same measured zone in the sample disc
Different cell samples.
Wherein, during the positioning table horizontal movement, according to the control of the horizontal displacement signal of the positioning table
Positioning table is continued to move to or stopped.
The method that the control positioning table is continued to move to or stopped includes：Calculate the horizontal displacement of the positioning table with
The difference of displacement is set, judges the difference whether in setting difference range；If in setting difference range, controlling institute
State positioning table and stop movement；Otherwise, the positioning table is controlled to continue to move horizontally.
Further, the measuring method of cyto-mechanics of the present invention also includes：The positioning table is arranged on displacement platform, controlled
The displacement platform level and/or vertical movement are made, the measuring unit is contacted with cell sample to be measured；In a measured zone
After measurement terminates, control bit moving stage level and/or vertical movement make the measuring unit measure the different surveys in the sample disc
Measure the cell sample in region.
During the displacement platform is moved horizontally, the displacement platform is controlled according to the horizontal displacement signal of the displacement platform
Continue to move horizontally or stop.Wherein, the control displacement platform is continued to move to or the method stopped and the positioning table
Control method is similar, will not be repeated here.
Preferably, the measuring method of cyto-mechanics of the present invention also includes：Collection shows the measuring unit and institute in real time
State the image of the position relationship of cell sample, so as to when there is deviation in the position of the measuring unit and cell sample can and
When adjust, it is ensured that the accuracy of metrical information.
The measuring method (as shown in Figure 6) of cyto-mechanics of the present invention is discussed in detail with a specific embodiment below.
By CCD observe and correspondingly move horizontally displacement platform, make first in probe perpendicular alignmnet region to be measured it is to be measured
The middle position of cell sample；Then control bit moving stage is moved in Z-direction, the cell sample to be measured is approached probe；Control
Piezoelectric ceramic stack drives probe to be moved in Z-direction, to adjust the exposure level of the probe and cell sample, obtains exposure level
The mechanics parameter of the cell sample in during change；After a cell sample measurement terminates, control positioning table is moved horizontally,
Make the next cell center to be measured of probe alignment (as shown in figure 3, scanning range can be 200 μm of 200 μ m, step pitch is 40 μm)；
Judge whether to arrive at the maximum scan scope of the positioning table, if repeating aforesaid operations without if, continue to measure same measurement
Different cell samples in region, otherwise, control bit moving stage level and/or vertical movement, with next measured zone
Cell sample is measured；Judge whether the scanning of completion Zone Full, otherwise the end operation if completing repeats above-mentioned behaviour
Make, continue to measure.
The measurement apparatus of cyto-mechanics of the present invention has the advantages that locating speed is fast, precision is high using positioning table, carries out X, Y
The quick and resetting of direction a small range, makes rapidly the middle position of the cell sample to be measured in probe alignment sample disc；
Have the advantages that moving range is big, precision is high, load capacity is strong using displacement platform, it is driven positioning table thereon and sample disc
Automatic switchover sample detection region, it is to avoid because positioning table moving range is small, the manual switching Sample Scan region disadvantage that time-consuming
End, easy to operate, measurement efficiency is high.
The preferred embodiment of the present invention is described in detail above in association with accompanying drawing, still, the present invention is not limited to above-mentioned reality
The detail in mode is applied, in the range of the technology design of the present invention, a variety of letters can be carried out to technical scheme
Monotropic type, these simple variants belong to protection scope of the present invention.
It is further to note that each particular technique feature described in above-mentioned embodiment, in not lance
In the case of shield, can be combined by any suitable means, in order to avoid unnecessary repetition, the present invention to it is various can
The combination of energy no longer separately illustrates.
In addition, various embodiments of the present invention can be combined randomly, as long as it is without prejudice to originally
The thought of invention, it should equally be considered as content disclosed in this invention.
Claims (14)
1. a kind of measurement apparatus of cyto-mechanics, it is characterised in that the measurement apparatus includes：
Sample disc (1), for placing cell sample；
Measuring unit (2), for being contacted with the cell sample in the sample disc (1), obtains the mechanics ginseng of the cell sample
Number；
Control unit (3), is connected with the measuring unit (2), for controlling the measuring unit (2) according to the mechanics parameter
Vertical movement, to change the exposure level of the measuring unit (2) and the cell sample, obtains the measuring unit (2)
The mechanics parameter of the cell sample in during the exposure level change；And
Positioning table (4), is connected with described control unit (3), and the sample disc (1) is provided with the positioning table (4), is used for
Controlled by described control unit (3), and drive the sample disc (1) level and/or vertical movement, so that the measuring unit
(2) cell sample in the sample disc (1) is measured；Control of the described control unit (3) to the positioning table (4) can using band
Adjust the PID closed-loop controls in dead band；
Wherein, the sample disc (1) includes multiple measured zones, and each measured zone is into the chessboard lattice distribution of dot matrix；
There is the promotion layer for promoting cell attachment, adjacent chessboard compartment has the suppression for suppressing cell attachment inside each gridiron pattern
Layer；
Wherein, the preparation of the sample disc comprises the following steps：
S1) a piece of template is provided, micron post is prepared in the template, makes chessboard of the micron post into dot matrix in template
Lattice distribution, forms the sample disc, wherein, one measured zone of multiple gridiron pattern formation, the sample disc includes multiple measurements
Region；
Wherein, preparing the method for micron post includes：Using coupled plasma etch template is reacted, poly- diformazan is utilized according to template
Radical siloxane carries out complex, obtains the micron post；
S2) sample disc is soaked in 10-100 μ g/mL collagen, laminin, fibronectin, gelatin, or
In 0.01% dextrorotation Poly-L-Lysine Solution, 37 DEG C are incubated 1 hour；
S3) using step S2) the micron post of the sample disc for preparing is seal, by western blot in hydrophobic plastics
The promotion layer for promoting cell attachment is modified with the surface of ware or glass dish substrate, the upper surface for making the micron post；And by institute
The plastic ware or glass dish substrate that are modified with the promotion layer for promoting cell attachment are stated in F-127, bovine serum albumin(BSA) or poly- second two
It is incubated 0.5 hour in alcoholic solution, makes that there is the inhibition layer for suppressing cell attachment between adjacent chessboard.
2. the measurement apparatus of cyto-mechanics according to claim 1, it is characterised in that described control unit (3) is according to institute
Stating mechanics parameter controls measuring unit (2) vertical movement to include：
Mechanics parameter acquired in the measuring unit (2) is compared by described control unit (3) with setting value, is used for：
In the mechanics parameter>During setting value, measuring unit (2) upward vertical movement is controlled, makes the measuring unit (2)
Weaken with the exposure level of the cell sample, the stop motion when mechanics parameter=setting value obtained again；
In the mechanics parameter<During setting value, measuring unit (2) vertical downward movement is controlled, makes the measuring unit (2)
Strengthen with the exposure level of the cell sample, the stop motion when mechanics parameter=setting value obtained again.
3. the measurement apparatus of cyto-mechanics according to claim 1, it is characterised in that the measuring unit (2) includes：
Laser (21), for launching light beam perpendicular to the sample disc (1)；
Detection module, can move vertically, for contacting and interacting with the cell sample in the sample disc (1), simultaneously
The light beam that the laser (21) is launched is reflected into the spot signal for characterizing mechanics parameter；And
Photoelectric conversion module (22), for receive the spot signal and be converted to characterize mechanics parameter electric signal and send extremely
Described control unit (3)；
Described control unit (3) connects the photoelectric conversion module (22) and detection module respectively, for receiving the sign power
The electric signal of parameter is learned, and controls the detection module to move vertically according to the mechanics parameter, to change the detection module
With the exposure level of the cell sample.
4. the measurement apparatus of cyto-mechanics according to claim 3, it is characterised in that the detection module includes：
Probe detector (23), for contacting and interacting with cell sample, while light beam is reflected into sign mechanics parameter
Spot signal；And
Piezoelectric ceramic stack (24), is connected with the probe detector (23)；For being controlled by described control unit (3), and drive
Probe detector (23) vertical movement.
5. the measurement apparatus of cyto-mechanics according to claim 3, it is characterised in that the measuring unit also includes：
Finely tune module, be connected with the photoelectric conversion module (22), for adjust the position of the photoelectric conversion module (22) with
Receive the spot signal.
6. the measurement apparatus of cyto-mechanics according to claim 3, it is characterised in that the measuring unit (2) also includes：
Image capture module (25), the image of the detection module and cell sample position relationship is shown for gathering in real time.
7. the measurement apparatus of cyto-mechanics according to claim 6, it is characterised in that the measurement apparatus also includes：
Positioning table (4), is connected with described control unit (3), and the sample disc (1) is provided with the positioning table (4), is used for
Controlled by described control unit (3), and drive the sample disc (1) level and/or vertical movement, so that the measuring unit
(2) the different cell samples of same measured zone in the sample disc (1) are measured.
8. the measurement apparatus of cyto-mechanics according to claim 7, it is characterised in that the measurement apparatus also includes：
Displacement platform (5), is connected with described control unit (3), and the positioning table (4) is provided with the displacement platform (5), is used for
Controlled by described control unit (3), positioning table (4) level and/or vertical movement are driven, so that the measuring unit (2)
Measure the cell sample of measured zones different in the sample disc.
9. a kind of usage right requires that the measurement apparatus of the cyto-mechanics any one of 1-8 performs the measurement of cyto-mechanics
Method, it is characterised in that the measuring method includes：
Cell sample is positioned in sample disc；
Measuring unit is contacted with the cell sample in the sample disc, obtain the mechanics parameter of the cell sample；
The measuring unit vertical movement is controlled according to the mechanics parameter, to change the measuring unit and the cell sample
Exposure level, the measuring unit is obtained the mechanics parameter of the cell sample in during the exposure level change；
The sample disc is installed on positioning table, the positioning table level is controlled and/or vertically moves, make the measuring unit
It is directed at cell sample to be measured；Control to the positioning table (4) can adjust the PID closed-loop controls in dead band using band；
The measuring method also includes：Before cell sample is positioned in sample disc, the sample disc is prepared, wherein, it is described
The preparation of sample disc comprises the following steps；
S1) a piece of template is provided, micron post is prepared in the template, makes chessboard of the micron post into dot matrix in template
Lattice distribution, forms the sample disc, wherein, one measured zone of multiple gridiron pattern formation, the sample disc includes multiple measurements
Region；
Wherein, preparing the method for micron post includes：Using coupled plasma etch template is reacted, poly- diformazan is utilized according to template
Radical siloxane carries out complex, obtains the micron post；
S2) sample disc is soaked in 10-100 μ g/mL collagen, laminin, fibronectin, gelatin, or
In 0.01% dextrorotation Poly-L-Lysine Solution, 37 DEG C are incubated 1 hour；
S3) using step S2) the micron post of the sample disc for preparing is seal, by western blot in hydrophobic plastics
The promotion layer for promoting cell attachment is modified with the surface of ware or glass dish substrate, the upper surface for making the micron post；And by institute
The plastic ware or glass dish substrate that are modified with the promotion layer for promoting cell attachment are stated in F-127, bovine serum albumin(BSA) or poly- second two
It is incubated 0.5 hour in alcoholic solution, makes that there is the inhibition layer for suppressing cell attachment between adjacent chessboard.
10. the measuring method of cyto-mechanics according to claim 9, it is characterised in that described according to the mechanics parameter
Controlling the method for the measuring unit vertical movement includes：
Mechanics parameter acquired in the measuring unit and setting value are compared：
In the mechanics parameter>During setting value, control the measuring unit upward vertical movement, make the measuring unit with it is described
The exposure level of cell sample weakens, the stop motion when mechanics parameter=setting value obtained again；
In the mechanics parameter<During setting value, control the measuring unit vertical downward movement, make the measuring unit with it is described
The exposure level enhancing of cell sample, the stop motion when mechanics parameter=setting value obtained again.
11. the measuring method of cyto-mechanics according to claim 9, it is characterised in that the measuring method also includes：
The sample disc is installed on positioning table, the positioning table level is controlled and/or vertically moves, make the measuring unit
It is directed at cell sample to be measured；
After a cell sample measurement terminates, the positioning table level and/or vertical movement are controlled, the measuring unit is surveyed
Measure the different cell samples of same measured zone in the sample disc.
12. the measuring method of cyto-mechanics according to claim 11, it is characterised in that in the positioning table horizontal movement
During, control the positioning table to continue to move to or stop according to the horizontal displacement signal of the positioning table.
13. the measuring method of cyto-mechanics according to claim 12, it is characterised in that the control positioning table after
Method that is continuous mobile or stopping including：
The difference of the horizontal displacement and setting displacement of the positioning table is calculated, judges the difference whether in setting difference range
It is interior；If in setting difference range, controlling the positioning table to stop movement；Otherwise, the positioning table is controlled to continue level
It is mobile.
14. the measuring method of the cyto-mechanics according to any one of claim 11-13, it is characterised in that the measurement
Method also includes：
The positioning table is arranged on displacement platform, the displacement platform level and/or vertical movement is controlled, makes the measuring unit
Contacted with the cell sample in region to be measured；
After a measured zone measurement terminates, control bit moving stage level and/or vertical movement make the measuring unit measurement institute
State the cell sample of the different measured zones in sample disc.
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