CN1172173C - Microscopic single cell sample cutting method and system - Google Patents
Microscopic single cell sample cutting method and system Download PDFInfo
- Publication number
- CN1172173C CN1172173C CNB011235020A CN01123502A CN1172173C CN 1172173 C CN1172173 C CN 1172173C CN B011235020 A CNB011235020 A CN B011235020A CN 01123502 A CN01123502 A CN 01123502A CN 1172173 C CN1172173 C CN 1172173C
- Authority
- CN
- China
- Prior art keywords
- cropping
- pin
- microactuator
- single cell
- base
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Abstract
The present invention belongs to the technical field of biology. In the present invention, the microscopic image of a tissue slice is transmitted to a computer through a CCD image sensor, and an interesting cell is outlined on the microscopic image by an input device; simultaneously, the position information of the outlined contour is obtained and stored by decoding the data packet of the input device by software; the computer controls a micro actuating mechanism to outline, cut and take out the contour outlined by the input device on the tissue slice. According to the shape of a single cell or a single type cell group, the present invention can make a cutting to obtain an ideal sample directly and accurately without the need of a heat plastic membrane so that the sample is conveniently processed.
Description
Technical field
The invention belongs to biological technical field, particularly be used for from microstructure section, obtaining single or the single type cell is the method for interested disease cell and the dedicated system of this method.
Background technology
Medical science develops into molecular level from cellular level, and the mankind are Fighting Disease fundamentally, at first must and assign a cause for an illness from the molecular level searching, just might study therapeutic scheme and methods of treatment then.And assign a cause for an illness from the molecular level searching, the first step is to obtain the disease cell sample.Medical research shows, in histotomy, and often only some or even several or cell generation pathology.For example, in the malignant tumor tissue section, the correlator group of canceration cell in early stage and cancerous tumor cell is in the visible visual field of microscope, usually less than 5%.Therefore, for the cause of disease information that obtains to determine reliably, must from section, obtain single or, be used for the molecular medicine analysis with a kind of sick cell.Present most of laboratory or laboratory are to operate at microscopically by manual, directly obtain with suction pipe, pin or blade; This method requires operating personnel's spirit high concentration, and action is quick especially, takes great energy time-consuming.In order to change this situation, have the people once to design various auxiliary body-micro-manipulators and come complete operation, but cell is very little, there is uncertainty again in the allogenic cell form, is difficult to prove effective by manual direct control micro-manipulator.
For being provided, a kind of reliable method obtains specific cell exactly, the National Cancer Institute of NIH (NIH) was in 1996, invented laser and obtained micro-dissections (LastrCapture Microdissection) device, as shown in Figure 1.Arcturun engineering corporation was developed to product in 1997.As shown in Figure 1, this installs by microscope 1, and laser system 2 and micro-dissections device 3 etc. is partly formed.Laser system 2 is installed in the microscope 1, and it is made up of semiconductor laser controller 4, semiconductor laser 5, collimating apparatus 6, lens 7 and catoptron 8, sees Fig. 2.The light path coaxial of the vertical optical path of laser beam and microscope 1.Micro-dissections device 3 is installed in the sample stage top of microscope 1, and it is made up of transferring arm 9, plastic cap 10 and transparent EVA hot plastic film 11, sees Fig. 3.Hot plastic film is attached on the plastic cap 10, and plastic cap is installed in the circular hole of transferring arm 9.The principle of work of this device is: fixed tissue slices 13 on the light surface glass 12, place on the objective table of microscope 1, find interesting areas, and the fine setting slice position, make interested cell be positioned at the center, visual field.Then, mobile transferring arm is placed on the interested tissue samples hot plastic film 11 on the plastic cap 10.Then, start laser controller 4, the laser that laser instrument 5 sends becomes parallel light beam after collimating apparatus 6, behind collimated light beam scioptics 7 and the catoptron 8, focuses on the interested cell, with hot plastic film 11 fusings that cover on the cell.The plastics that melted spread gradually and are penetrated in the space between tissue, and plastics cooling rapidly solidify histocyte, like this similar process of the process of embedding cell and paraffin-embedded tissue.Lift transferring arm 9, plastic cap 10 lifts thereupon.At this moment, sample tissue can be separated with histotomy 13 along with lifting of plastic cap 10 at polymkeric substance-organize the edge of complex to be torn.Because hot plastic film is attached on the plastic cap 10, when it lifts, hot plastic film is got up thereupon, and that tissue that covers when hot plastic film melted is torn from histotomy and is taken up, be that interested part cuts down from section, and be not melted the organizing still on glass sheet 12 of thermoplast infiltration, see Fig. 4.This just means, can be downcut by the tissue of laser illumination, and remaining intact tissue is harmless.The advantage of this method has: the integrality that 1) has kept being cut cellular morphology; 2) fast and reliable.More than 3 years, they have been developed to LCM-210,220, products such as UPGD-200 and FLUOR-300.This method cutting size depends on the size that focuses on circular laser facula., sick cell does not have given shape often, and this method is difficult to cut according to the specific modality of cell or tissue.Sometimes, need analyze the similar cell in big zone, this method can only be leaned on repeatedly to cut and realize that efficient is low.And, although thermoplast makes troubles but can handle to sample analysis to analyzing not influence.
Summary of the invention
The objective of the invention is to overcome the deficiency of above-mentioned technology, a kind of single cell micro-cutting method and system thereof of novelty is provided.This method can be cut according to the shape of individual cells or single type cell mass.Obtain desirable sample, obtain directly and accurately, do not need hot plastic film again, convenience sample is handled.
The present invention proposes a kind of microscopic single cell sample cutting method, it is characterized in that:
1) microscopic image with observed histotomy in the microscope is sent to computing machine by digital CCD image sensor, and shows on display:
2) utilize input equipment, on the microscopic image of shown histotomy, touch off interested cell; Meanwhile, carry out software decode, obtain the positional information of institute's depiction in outline and store by packet to input equipment;
3) computing machine is defeated by the microactuator controller with the positional information of institute's depiction in outline, and handles under the control of computing machine, and the signal controlling microactuator after the processing comes out input equipment institute depiction in outline cropping on histotomy;
4) the cell cutting of institute's cropping on the histotomy is taken out.
Above-mentioned said input equipment is the mouse of buying on the market, or board, or light pen; Said microactuator mainly is made up of bidimensional flexible hinge driver and cropping pin.
The present invention proposes a kind of individual cells or single micro-dissections system of planting cell that adopts above-mentioned cutting method, comprise microscope, digital ccd image sensor, input equipment and computing machine, it is characterized in that, also comprise microactuator pillar, microactuator, microactuator controller, and the positional information of the profile of input equipment institute cropping is obtained program; Said digital ccd image sensor is installed on the microscopical microimaging interface, directly links to each other with computing machine; Said microactuator pillar is fixed on the Microscope base; Said microactuator is installed on the saddle of pillar; Said microactuator controller is handled microactuator.
Said microscope is an inverted microscope;
On the said microactuator pillar cam mechanism and saddle are arranged, rotate said cam mechanism and the microactuator that is installed on the saddle is risen or fall;
Said microactuator is made up of the flexible topworks of bidimensional base, bidimensional flexible hinge driver, cropping pin pedestal, cropping pin base, cropping needle stand, cropping pin and cropping pin briquetting; Said bidimensional flexible hinge driver is installed on the said topworks base, this base is installed on the saddle of this pillar, not with microscopical movable workbench, when the cell in the big zone of needs cutting, the cropping pin is motionless, can move microscopical worktable and carry out cropping; Said bidimensional flexible hinge driver is driven by electrostriction element, can upwards produce 100 μ m displacements in two power of X, Y; Said cropping pin pedestal is installed on the bidimensional flexible hinge driver, and an end is connected with the stiff end of this driver, and the other end is a free end; The free end of said cropping pin pedestal has a square opening, and cropping pin base is installed on the square hole; The square projection of said cropping pin base inserts in the square hole of pedestal; Have steel ball to support between said cropping pin base and the cropping pin pedestal, base can be with respect to the pedestal free movement; Said cropping needle stand is fixed on the cropping pin base, and said cropping pin is installed on the cropping needle stand, is fixed with briquetting; Said cropping pin becomes 30-60 ° of angle with surface level.
Said cropping pin is a stainless pin, and needle point is less than 0.5 μ m;
Said microactuator controller is made up of data register, D/A converter and high-voltage amplifier, and X is identical with the Y two-way; Said register is deposited the positional information that computing machine is sent here; Said D/A converter converts the positional information of depositing in the register to analog voltage signal; Said high-voltage amplifier zooms into high pressure with the voltage signal of D/A converter output, removes to drive the electrostriction executive component;
Said electrostriction element is by about 1 centimetre of the length of side, and thickness is that the ternary system piezoelectric ceramics sheet of 1-2 millimeter builds up, and it is 2-3 centimetre that an insulating trip, total length are arranged between two; The positive pole of said each piezoelectric ceramic piece and negative pole are towards consistent, and positive and negative electrode is connected in parallel respectively.
Said high-voltage amplifier is formed Darlington formula amplifier by three transistors;
Said input equipment is the mechanical type mouse, and friction ball wherein is through refining, and the circularity height is avoided " skidding " phenomenon (be that mouse moves, but do not produce positional information);
The positional information of said input equipment depiction in outline is to adopt serial mouse microsoft coded format, is undertaken by the packet to mouse that software decode obtains.
Adopt described cutting individual cells or single course of work of planting little diced system of cell, may further comprise the steps:
1) histotomy is fixed on the light surface glass plate, is placed in again on the microscopical worktable, and observe, find interested cell by eyepiece;
2) interested cell image is presented on the display of computing machine by digital ccd image sensor;
3) utilize input equipment that the profile cropping of interested cell or cell mass is come out, carry out software decode by packet simultaneously, obtain the positional information of institute's depiction in outline and store input equipment; If the profile of selected cell or institute's cropping is undesirable, can reselect and cropping, the positional information of storage is also along with refreshing;
4) after the profile satisfaction of institute's cropping, again by the positional information input microactuator controller of computing machine, and control and carry out D/A switch and the signal of changing is carried out the high pressure amplification storage, remove to drive microactuator; Microactuator is handled a cropping pin, and the profile according to input equipment institute cropping on histotomy touches off interested cell;
5) after finishing cropping on the histotomy, rotate the cam mechanism on the column, rise microactuator, a kind of xanthan gum is sticked on the cropping needle point, then tip position is transferred to the center of institute's cutting zone, and rotating cam mechanism, fall microactuator; Be rotated further cam mechanism immediately, rise microactuator, just the cell that is cut has been stained with, i.e. cutting is interested cell down;
6) take off sample from the cropping needle point, with the glue on the alcohol removing needle point, for operation next time is ready to.
Zone if desired is at 100 * 100 μ m
2More than, can directly move microscopical worktable and carry out cropping, at this moment the cropping pin is motionless, other is identical with above-mentioned steps.
This single cell micro-dissections provided by the invention system, it can cut according to the profile of cell or required region contour, does not need thermoplastic film, have conveniently, flexibly, selectivity is strong, helps follow-up advantages such as sample process.
Description of drawings
Fig. 1 is existing single cell micro-dissections system architecture synoptic diagram.
Fig. 2 is the laser system structural representation in the existing diced system.
Fig. 3 is the micro-dissections device structural representation in the existing diced system.
Fig. 4 is existing single cell micro-dissections schematic diagram.
Fig. 5 is microscopic single cell sample cutting method of the present invention and device synoptic diagram thereof.
Fig. 6 is a microactuator pillar embodiment synoptic diagram of the present invention.
Fig. 7 is a microactuator embodiment synoptic diagram of the present invention.
Fig. 8 is a bidimensional flexible hinge formula driver embodiment synoptic diagram of the present invention.
Fig. 9 is an electrostriction executive component example structure synoptic diagram of the present invention.
Figure 10 is cropping pin embodiment drive principle figure of the present invention.
Figure 11 is the process flow diagram that the serial mouse packet is decoded of the present invention.
Figure 12 is a microactuator controller embodiment block scheme of the present invention.
Figure 13 is high-voltage amplifier embodiment circuit theory diagrams of the present invention.
Embodiment
Below in conjunction with accompanying drawing the embodiment that adopts single cell micro-cutting method of the present invention and system thereof is illustrated.
The system that the embodiment of a kind of single cell micro-dissections system of the present invention design is made up of unit such as inverted microscope, digital ccd image sensor, microactuator pillar, microactuator, microactuator controller, input equipment (present embodiment is the mechanical type mouse) and computing machines, and carry out software decode by packet to mouse, the positional information of obtaining institute's cropping is carried out the method for single cell micro-dissections, as shown in Figure 5.Wherein, the inverted microscope of microscope 14 for buying on the market, digital ccd image sensor 15, mouse 22 and computing machine 23 are also bought from the market; Microactuator pillar 16 is made up of column 24, camshaft 25, handwheel 26, handwheel set nut 27, bearing 28, bearing holder (housing, cover) 29, cam 30, saddle 31, locating piece 32 and camlock nut 34, sees Fig. 6; Microactuator 17 is made up of two little flexible topworks bases 36, bidimensional flexible hinge driver 38, cushion block 39, cropping pin pedestal 40, steel ball 42, cropping pin base 43, cropping needle stand 44, cropping pin 45, cropping pin briquetting 46, spring base 49, first spring 50 and first steel ball 51, sees Fig. 7; Microactuator controller 20 is made up of data register, D/A converter and high-voltage amplifier etc., sees Figure 12.The annexation of each parts is: pillar 16 usefulness screws 21 are fixed on the base of microscope 14; Microactuator 17 is installed on the saddle of pillar 16, and the input of microactuator controller 20 links to each other with computing machine 23, and output links to each other with microactuator 16; Numeral ccd image sensor 15 is installed on the microimaging interface of microscope 14, links to each other with computing machine 23.
Single cell micro-cutting method embodiment of the present invention specifically may further comprise the steps:
1) histotomy 19 for preparing is fixed on the object carrier sheet 18, and is placed on the worktable of microscope 14; Observe by eyepiece, adjust focal length, make the image of histotomy 19 can observe the most clearly, adjust the position of digital ccd image sensor 15 again, make on display observed image also the most clear; The worktable of mobile again microscope 14 is observed from microscope, seeks needed cell; After needed cell finds, it is adjusted to the center, visual field of microscope 14;
2) then, with the interested cell outline of mouse 22 croppings, simultaneous computer 23 carries out software decode by the packet to mouse, obtains and store the positional information of institute's depiction in outline, if the profile cropping is dissatisfied, cropping again is till satisfaction;
3) by handwheel 26 rotating cams 30 on the pillar 16, fall microactuator 17, the needle point that is about to cropping pin 45 drops on the outline line of institute's cropping; Operational computations machine 23 is defeated by microactuator controller 20 with the positional information of 22 croppings of mouse; Register in little execution mechanical control device 20 stores away earlier after receiving the positional information of computing machine 23 output, is defeated by D/A converter again; D/A converter in time converts digital position information to simulating signal and is defeated by high-voltage amplifier, and the output of high-voltage amplifier connects electrostriction element 52 or 53; Electrostriction element 52 or 53 produces and stretches under the output voltage effect of high-voltage amplifier, thereby drives 43 motions of cropping pin base, and cropping pin 45 movement therewith are promptly carried out cropping on histotomy 19;
4) finish cropping after, the handwheel 26 on the rotation post 16 rotates cam 30, rises microactuator 17, will be stained with on the needle point that glue is put on cropping pin 45; Observation display, the worktable of mobile microscope 14 makes under the needle point that is centered close to cropping pin 14 in zone of cropping; Handwheel 26 on the rotation post 16 rotates cam 30, falls microactuator 17, even cropping pin 45 drops on the center in institute cropping zone; Then, rotate handwheel 26 again, microactuator 17 is risen, cropping pin 45 has been stained with the cell of cropping thereupon, promptly cuts out;
5) cell under will cutting with specific purpose tool takes off, and is placed in the little centrifuge tube, with pending, with alcohol cropping pin 45 thing needle points is wiped clean simultaneously, for operation next time is ready to.
Because microactuator 17 is mounted on the pillar 16, keep independent with the worktable of microscope 14, when needed zone greater than 100 * 100 μ m
2The time, can cropping pin 45 motionless, as long as travelling table just can be realized cropping.
The present embodiment each several part is formed and function is described in detail as follows:
The microactuator pillar of present embodiment as shown in Figure 6, it is made up of column 24, camshaft 25, handwheel 26, handwheel set nut 27, bearing 28, bearing holder (housing, cover) 29, cam 30, microactuator saddle 31, locating piece 32, locating piece gib screw 33 and camlock screw 34.Bearing 28 is contained in earlier on the camshaft 25, between two bearings bearing holder (housing, cover) 29 is arranged, and refills on column 24, and cam 30 and handwheel 26 are contained on the camshaft 25, and uses set nut 34 and 27 with its locking respectively, makes it to rotate with axle 25.Microactuator saddle 31 is contained on the column 24, and its column part inserts in the center hole of column 24, contacts with cam 30, can move up and down.Column 24 sides and dead eye are the assembly technology holes to a circular hole should be arranged, and there is a rectangular slot upper end, are the locating slot of microactuator 17.Block 32 is fixed on the cylindrical mesa of microactuator saddle 31 by screw 33, drops in the rectangular slot on the column 24, prevents that microactuator saddle 31 from rotating, and prevents that promptly microactuator 17 from rotating, and guarantees cropping pin 45 accurate croppings.Rotate handwheel 26, cam 30 rotates thereupon, when the peak of cam 30 up the time, rises microactuator saddle 31, the arrival extreme higher position, on the contrary fall.
The microactuator of present embodiment as shown in Figure 7, it is made up of the flexible topworks of bidimensional base 36, bidimensional flexible hinge driver 38, backing plate 39, cropping pin pedestal 40, steel ball 42, cropping pin base 43, cropping needle stand 44, cropping pin 45, cropping pin briquetting 46, spring base 49, first spring 50, first steel ball 51 and gib screw 35,37,41,47 and 48.The flexible topworks of bidimensional base 36 is fixed on the microactuator saddle 31 by screw 35.Bidimensional flexible hinge driver 38 is fixed on the flexible topworks of the bidimensional base 36 by screw 37.Cropping pin pedestal 40 is fixed on the stiff end of bidimensional flexible hinge driver 38 by screw 41, and cushion block 39 is arranged between the two, prevents that free end from interfering with each other.The free end of cropping pin pedestal 40 has a square opening, and the square projection of cropping pin base 43 inserts in the square hole; On the square important actor one side of cropping pin base 43 3 uniform fossettes are arranged, steel ball 42 embeds fossette, places on the cropping pin pedestal 40, and carrying cropping pin base 43 can be free to slide cropping pin base 43.Cropping needle stand 44 has an inclined-plane, with horizontal plane angle 30-60 °, a stria is arranged on the inclined-plane, and is in cropping pin 45 mounting grooves, above briquetting 46 is pressed in, fixing with screw 47.Spring base 49 usefulness screws 48 are fixed on the free end of cropping pin pedestal 40, one end of first spring 50 is placed in the circular port of spring base 49, last first steel ball 51 in other end top, first steel ball 51 contacts with a face of square projection at the bottom of the cropping pin again, another corresponding surface of square projection contacts (two contacts of X, Y are arranged, and this contact is the X contact) with a contact of two-dimension flexible hinge driver 38.When bidimensional flexible hinge driver 38 under the effect of electrostriction element 52 and 53 (set 52 here for directions X, 53 is the Y direction), its contact moves right, thereby drive cropping pin 45 cropping to the right, head on first steel ball, 51 compressions, first spring 50 simultaneously, otherwise contact is to left movement, 50 elongations of first spring, head on first steel ball 51, pushing away cropping pin base 43 and contact, promptly pushing away cropping pin 45 cropping left synchronously to left movement, Y to motion class seemingly, do not give unnecessary details here.
The bidimensional flexible hinge driver of present embodiment is shown in Fig. 8 (a), and it is made up of bidimensional flexure hinge mechanism 54 and electrostriction element 52,53.Tiltedly the shadow part is removable among the figure, and other parts maintain static.When electrostriction element 52 extends, promote BD to left movement under the effect of high-voltage amplifier output voltage.The A point is motionless, is fulcrum, and the B point is the application point of power, promotes the C point to left movement.Simultaneously, the E point is motionless to be fulcrum, and the D point is to left movement, and the F point just moves right, and along with the M point also moves right, the F point is not only moving point, and as fulcrum, the C point is to left movement, and contact M just moves right.Therefore, this mechanism has the displacement amplification, and mechanics principle is seen Fig. 8 (b).Otherwise when electrostriction element 52 stretched, the M point was to left movement.In like manner, when electrostriction element 53 elongations, contact N moves (being as the criterion towards paper with the people) backward, otherwise travels forward.
The electrostriction element of present embodiment as shown in Figure 9, it is made up of piezoelectric ceramic piece 55 and insulating trip 56.The material of piezoelectric ceramic piece 55 is a ternary system, about 1 centimetre of the length of side, square, thick 1-2 millimeter.Insulating trip 56 is a phenolic board, thick 0.1 millimeter, and all piezoelectric ceramics positive and negative electrodes are towards unanimity, and are connected in parallel.
The cropping pin drive principle of present embodiment as shown in figure 10.When contact M moves right (referring to Fig. 8) under the effect of electrostriction element 52, promote cropping pin base 43 and move right, cropping pin base 43 heads on first steel ball, 51 compressions, first spring 50 again.Otherwise contact is to left movement, and 50 elongations of first spring head on first steel ball 51, promote cropping pin base 43 and contact M synchronously to left movement.Cropping pin base 43 left, right motion, the also synchronous left and right motion of cropping pin 45 mounted thereto.In like manner, when contact N motion (being as the criterion towards paper with the people) backward under electrostriction element 53 effects, pushing away also motion backward of cropping pin base 43, cropping pin base 43 also heads on second steel ball, 59 compressions, second spring 58.Otherwise contact N travels forward, and second steel ball 59 is withstood in 58 elongations of second spring, is pushing away cropping pin base 43 and is travelling forward synchronously with contact N.Cropping pin base 43 forward and backward moving, the cropping pin 45 that is installed on it is also forward and backward thereupon to be moved.But contact M and N be independently moving both, can be synchronized with the movement again, guarantees that cropping pin 45 can be along the profile cropping of mouse cropping.
The flow process that the serial mouse packet is decoded of present embodiment as shown in figure 11.In the Microsoft standard, the asynchronous communication parameter of serial mouse comprises baud rate, 7 bit data and 1 position of rest, and data packet format is as shown in the table:
| Byte | D 7 | D 6 | D 5 | D 4 | D 3 | D 2 | D 1 | D 0 |
| 1 | X | 1 | LB | RB | Y 7 | Y 7 | X 7 | X 6 |
| 2 | X | 0 | X 5 | X 4 | X 3 | X 2 | X 1 | X 0 |
| 3 | X | 0 | Y 5 | Y 4 | Y 3 | Y 2 | Y 1 | Y 0 |
In the table, LB represents left button state (" 1 " illustrates and connects down), and RB represents right button state (" 1 " illustrates and connects down), X
7-X
0Then relatively last time coordinate X movement value (X
7Be sign bit), Y
7-Y
0Be the Y movement value (Y of relative last time of coordinate
7Be sign bit).
Mouse data adopts three frame host-host protocols, and each frame data is 8 bits.Present embodiment adopts interrupt mode work, whenever calls once to interrupt, and the microprocessor of mouse will send three frame informations, and the current state of mouse is passed to computing machine.
After computing machine 13 is finished initialization, set baud rate, normally 1200 bps, and open interruption, prepare to receive data.After computing machine 13 receives first frame data, judge whether correct; If correct, each frame data is handled, take out X
7, X
6, Y
7, Y
6Then, judge that receiving second frame data does not have; If receive, whether correct judgment again, if correct, handles again.Take out X
5-X
0In like manner, Y
5-Y
0After finishing processing, with data X
7-X
0, Y
7-Y
0Store.
The composition of the microactuator controller of present embodiment as shown in figure 12, it is made up of data register, D/A converter and high-voltage amplifier.Positional information by computing machine 13 outputs deposits data register earlier in, imports D/A converter again; D/A converter converts analog quantity to being about to digital quantity, and the input high-voltage amplifier amplifies.Signal is passed to electrostriction element 52 and 53 after amplifier amplifies, control elongating or shortening of they.
The high-voltage amplifier circuit structure of present embodiment as shown in figure 13, it is by resistance R 1, R2 and R3, low voltage transistor T1, T2 and high voltage transistor T3 form.T1 and T2 form Darlington transistor, and the D/A converter output signal is amplified, and guarantee that T3 has bigger base current, make T3 can be operated in linear zone.
Claims (10)
1, a kind of microscopic single cell sample cutting method is characterized in that:
1) microscopic image with observed histotomy in the microscope is sent to computing machine by digital CCD image sensor, and shows on display:
2) utilize input equipment, on the microscopic image of shown histotomy, touch off interested cell; Meanwhile, carry out software decode, obtain the positional information of institute's depiction in outline and store by packet to input equipment;
3) computing machine is defeated by the microactuator controller with the positional information of institute's depiction in outline, and handles under the control of computing machine, and the signal controlling microactuator after the processing comes out input equipment institute depiction in outline cropping on histotomy;
4) the cell cutting of institute's cropping on the histotomy is taken out.
2, microscopic single cell sample cutting method as claimed in claim 1 is characterized in that: said input equipment is a mouse, or board, or light pen is a kind of.
3, microscopic single cell sample cutting method as claimed in claim 2, it is characterized in that: said input equipment is the mechanical type mouse, the positional information of said input equipment depiction in outline is to adopt serial mouse microsoft coded format, is undertaken by the packet to mouse that software decode obtains.
4, a kind of single cell sample micro-dissections system that implements the described method of claim 1, comprise microscope, digital ccd image sensor, input equipment and computing machine, it is characterized in that, also comprise microactuator pillar, microactuator, microactuator controller, and the positional information of the profile of input equipment institute cropping is obtained program; Said digital ccd image sensor is installed on the microscopical microimaging interface, directly links to each other with computing machine; Said microactuator pillar is fixed on the Microscope base; Said microactuator is installed on the pillar; Said microactuator controller is handled microactuator.
5, single cell sample micro-dissections as claimed in claim 4 system is characterized in that said microscope is an inverted microscope.
6, single cell sample micro-dissections as claimed in claim 4 system is characterized in that said input equipment is the mechanical type mouse.
7, single cell sample micro-dissections as claimed in claim 4 system is characterized in that on the said microactuator pillar cam mechanism and saddle being arranged, and rotates said cam mechanism and the microactuator that is installed on the saddle is risen or falls.
8, single cell sample micro-dissections as claimed in claim 7 system, it is characterized in that said microactuator is made up of the flexible topworks of bidimensional base, bidimensional flexible hinge driver, cropping pin pedestal, cropping pin base, cropping needle stand, cropping pin and cropping pin briquetting; Said bidimensional flexible hinge driver is installed on the said topworks base, and this base is installed on the saddle of this pillar.
9, single cell sample micro-dissections as claimed in claim 8 system, it is characterized in that, said bidimensional flexible hinge driver is driven by electrostriction element, said cropping pin pedestal is installed on the bidimensional flexible hinge driver, one end is connected with the stiff end of this driver, and the other end is a free end; The free end of said cropping pin pedestal has a square opening, and cropping pin base is installed on the square hole; The square projection of said cropping pin base inserts in the square hole of pedestal; Have steel ball to support between said cropping pin base and the cropping pin pedestal, base can be with respect to the pedestal free movement; Said cropping needle stand is fixed on the cropping pin base, and said cropping pin is installed on the cropping needle stand, compresses with briquetting; Said cropping pin becomes 30-60 ° of angle with surface level; Said cropping pin is a stainless pin, and needle point is less than 0.5 μ m.
10, single cell sample micro-dissections as claimed in claim 4 system is characterized in that said microactuator controller is made up of data register, D/A converter and high-voltage amplifier, and X is identical with the Y two-way; Said register is deposited the positional information that computing machine is sent here; Said D/A converter converts the positional information of depositing in the register to analog voltage signal; Said high-voltage amplifier zooms into high pressure with the voltage signal of D/A converter output, removes to drive the electrostriction executive component.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011235020A CN1172173C (en) | 2001-07-26 | 2001-07-26 | Microscopic single cell sample cutting method and system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011235020A CN1172173C (en) | 2001-07-26 | 2001-07-26 | Microscopic single cell sample cutting method and system |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1332365A CN1332365A (en) | 2002-01-23 |
| CN1172173C true CN1172173C (en) | 2004-10-20 |
Family
ID=4665106
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB011235020A Expired - Fee Related CN1172173C (en) | 2001-07-26 | 2001-07-26 | Microscopic single cell sample cutting method and system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1172173C (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE10342264C5 (en) * | 2003-09-12 | 2012-10-31 | Leica Biosystems Nussloch Gmbh | System for uniquely assigning histological cassettes and slides |
| JP2010502983A (en) * | 2006-09-06 | 2010-01-28 | コン，ジアン−チエン | Method and apparatus for cutting out fresh tissue slices |
| CN101354240B (en) * | 2008-07-15 | 2010-06-02 | 南京航空航天大学 | Micro nano-scale fiber high precision measuring method based on optical microscopy |
| US8995733B2 (en) * | 2009-04-28 | 2015-03-31 | Koninklijke Philips N.V. | Microdissection method and information processing system |
| US8744163B2 (en) * | 2010-02-09 | 2014-06-03 | International Genomics Consortium | System and method for laser dissection |
| CN103959036A (en) * | 2011-11-25 | 2014-07-30 | 奥林巴斯株式会社 | Tissue segmentation apparatus, cell sorting apparatus, cell sorting system, tissue display system, substrate, extendible member, tissue segmentation method, and cell sorting method |
| CN102607880B (en) * | 2012-03-23 | 2014-08-27 | 苏州大学 | Piezoelectric micro-dissection system, dissection depth positioning method and dissection method |
| WO2014184005A1 (en) * | 2013-05-15 | 2014-11-20 | Koninklijke Philips N.V. | Tissue separation from a sample |
| US10000732B2 (en) * | 2015-11-20 | 2018-06-19 | National Health Research Institutes | Microfluidic dual-well device for highthroughput single-cell capture and culture |
| CN111474172B (en) * | 2020-04-20 | 2023-03-24 | 中国人民解放军陆军军医大学第一附属医院 | Target cell acquiring apparatus and target cell acquiring method |
-
2001
- 2001-07-26 CN CNB011235020A patent/CN1172173C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN1332365A (en) | 2002-01-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1172173C (en) | Microscopic single cell sample cutting method and system | |
| CN108441417B (en) | Bacterial colony picking instrument | |
| US9891143B2 (en) | Cylinder tip mounting head, and head device and movement device each using same | |
| US20060141616A1 (en) | Cell colonies dissecting and transplanting apparatus | |
| CN101294953B (en) | Motor cell real-time tracing system and method | |
| KR20140033340A (en) | Devices, systems, and methods for extracting a material from a material sample | |
| CN103235407A (en) | Multi-sample stage allowing for automatic movement of inverted microscopes | |
| JPWO2017061387A1 (en) | Sampling system | |
| CN201931205U (en) | Double-station laser etching machine for touch screen ITO (indium tin oxide) thin films | |
| WO2018193719A1 (en) | Cell transfer device and cell transfer method | |
| CN1496799A (en) | Cutting method of prittle material base plate, its devive and processing device | |
| CN110669653B (en) | Single-cell automatic printing system | |
| JP2000171360A (en) | Colony gathering method and device therefor | |
| CN102607880B (en) | Piezoelectric micro-dissection system, dissection depth positioning method and dissection method | |
| CN109100189B (en) | Cell tissue sample film-making device | |
| CN1306308C (en) | Microscope stage | |
| CN112295625A (en) | Fine pipetting mechanical arm and operating system thereof | |
| CN102179831B (en) | Micro-channel processing equipment of microfluidic chip | |
| US6908766B2 (en) | Device and method for capturing biological tissues | |
| EP3730598A1 (en) | Apparatus for treating biological material | |
| CN103278365B (en) | Drum type biochip spotting device and method thereof | |
| CN203337434U (en) | Roller type biochip sample application device | |
| CN214529017U (en) | Novel accurate positioning platform of slide glass | |
| CN202485924U (en) | Piezoelectric microdissection system | |
| CN2673597Y (en) | Splinter |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |