CN106646845A - Microscopic system for realizing automatic focusing and three-dimensional reconstruction and image obtaining method thereof - Google Patents

Abstract

The invention provides a microscopic system for realizing automatic focusing and three-dimensional reconstruction and an image obtaining method thereof. The microscopic system comprises a PC; a microscopic camera, in front of which a microscopic lens is arranged; a single-chip microcomputer, which is used for receiving a command signal of the PC, converting the command signal into a control signal and outputting the control signal; a first stepping motor, which receives circuit board control and is used for driving a first platform to rotate; a second stepping motor, which receives circuit board control and is used for driving the first platform to move back and forth; a third stepping motor, which receives circuit board control and is used for driving the first platform to adjust inclination angle alpha; a fourth stepping motor, which receives circuit board control and is used for driving the first platform to move up and down; and a fifth stepping motor, which receives circuit board control and is used for driving the first platform to move back and forth. The microscopic system realizes automatic focusing and three-dimensional reconstruction through the image obtaining method; the system is simple in circuit and optical path design and is high in integration; and man-made errors can be reduced.

Description

Realize the microscopic system and its image acquiring method of auto-focusing and three-dimensional reconstruction

Technical field

The invention belongs to micro-vision signal detection, process and control field, and in particular to one kind realize auto-focusing and The microscopic system and its image acquiring method of three-dimensional reconstruction.

Background technology

With the development of medical instrument, microscopical amplification in modern medicine instrument microscopic system is increasing to be led Cause the depth of field less and less, therefore the difficulty of manual focusing will necessarily be increased using manual focusing.In prior art, simple is automatic Focusing system has been implemented, and its system mainly includes PC and corresponding stepping motor control circuit, and wherein PC is mainly completed Micrograph image processing, circuit for controlling motor are controlled according to the signal that PC feeds back to motor.And show in modern medicine The three-dimensional reconstruction of object under micro- vision, commercially also there is the product of comparative maturity, and such as the production of Japanese Olympus company is sharp Light Laser Scanning Confocal Microscope can realize that the image after the three-dimensional reconstruction to cell image, the three-dimensional reconstruction is pseudo-colourss graphics Picture, is not 3-D view truly；It is right that the stereomicroscope of German Carl Zeiss Inc.'s production can also be realized The three-dimensional reconstruction of cell image, the essence of its operation principle is that two monocular tube microscopes are placed side by side, the optical axis of two lens barrels The visual angle that formed when constituting equivalent to people with one object of binocular vision, with this three-dimensional stereogram is formed Picture, but its zooming section still adopts and manually carries out.In existing product technology, be capable of achieving automatic focusing can not be real The three-dimensional reconstruction of existing image, is capable of achieving 3-dimensional reconstruction, not the function with automatic focusing.

The content of the invention

Present invention seek to address that technical problem present in prior art, and propose that one kind can realize that micro- object is automatic Focusing can realize the microscopic system of 3-dimensional reconstruction again.

For achieving the above object, it is proposed by the present invention to realize that auto-focusing and the microscopic system of three-dimensional reconstruction include PC； Microimaging head, with PC communication connection, the microimaging head front is provided with microlens, and microlens will be waited to see Examine the micro- amplification of object；Single-chip microcomputer, communicates to connect with the PC and receives the command signal of PC and be converted into control signal Output；A number motor, is connected and for driving a platform to rotate with a platform；No. two motors, It is connected with a platform and for driving a platform to move forward and backward；No. three motors, with a platform Connect and for driving a platform inclination angle [alpha]；No. four motors, are connected and for driving with a platform A number platform is moved up and down；No. five motors, be connected with a platform and for driving a platform before After move；A number step motor drive plate, is connected and according to the control signal control of single-chip microcomputer output with the single-chip microcomputer Number motor work；No. two step motor drive plates, are connected and according to the control of single-chip microcomputer output with the single-chip microcomputer Signal processed controls No. two motors work；No. three step motor drive plates, are connected and according to the monolithic with the single-chip microcomputer The control signal of machine output controls No. three motors work；No. four step motor drive plates, are connected and root with the single-chip microcomputer The control signal exported according to the single-chip microcomputer controls No. four motors work；No. five step motor drive plates, with the monolithic Machine connects and controls No. five motors work according to the control signal of single-chip microcomputer output.

Object to be seen disposed thereon is driven to be rotated by number platform of a step motor drive, it is possible to obtain Micro-image under different rotary angle conditions, therefore come the binocular work(in the middle of simulated implementation three-dimensional reconstruction by way of rotation Can, therefore when angle rotation is carried out, rotated frequently with the mode plus 180 ° on the basis of being rotated, so Just can be with the left view in the middle of simulated implementation binocular three-dimensional reconstruction and the acquisition function of right view, by No. two step motor drive Number platform drives object to be seen disposed thereon to be moved forward and backward, before and after can adjusting micro- object in the middle of the visual field Position；Number platform of No. five motor Motor drives drives object to be seen disposed thereon to be moved left and right, and can adjust Right position of the whole micro- object in the middle of the visual field；Driven by number platform of No. three step motor drive and disposed thereon wait to see Examine object and carry out angle of inclination a and be adjusted, reach and realize entering the not ipsilateral of micro- object by inclining different angles Row observation；Number platform of No. four step motor drive drives object to be seen disposed thereon to move up and down, and reaches to The platform at the adjusted good anglec of rotation and angle of inclination carries out automatic focusing；Microlens on No. 1 platform to treating micro- object It is amplified, is then imaged on afterwards on the sensitive chip in the middle of photographic head, finally transmit is carried out at image to the software of PC Reason, reaches the purpose of three-dimensional modeling, and the software that can pass through in addition on operation PC realizes the control worked each motor, The purpose of auto-focusing is reached, such that it is able to reduce the error for artificially causing, that is, realizing automatic focusing and have can realize Three-dimensional Gravity Build.

Further, realize that micro- object auto-focusing and the microscopic system of three-dimensional reconstruction also include No. two platforms and No. three Platform a, motor is fixedly arranged on No. two platforms and drives a platform to rotate, No. three steppings Motor is arranged between No. two platforms and No. three platforms and drives No. two platforms to drive a platform inclination, No. two steps Stepper motor, No. four motors and No. five motors are connected with No. three platforms and drive No. three platforms drive described No. two Platform and a platform are moved forward and backward respectively, move up and down and moved left and right.

Further, because the field range of micro-vision is smaller, thus tilt angle alpha meet -5 ° >=α >=5 ° can be with Obtain satisfied visual effect.

It is above-mentioned to realize that auto-focusing and the microscopic system micro-image acquisition methods of three-dimensional reconstruction include step：

S10：The demarcation of photographic head, the plane reference method proposed using Zhang Zhengyou carries out the demarcation of photographic head；

S20：The acquisition of left view and right view, a certain frame of video signal is used as left view after automatic focusing is completed Figure, a certain frame of video signal is used as right view after the anglec of rotation is completed；

S30：Image characteristics extraction and matching, characteristic point is extracted using GFTT algorithms, using SIFT algorithm conducts The algorithm of Image Feature Matching；

S40：Three-dimensional reconstruction process, using the method for feature based zone broadening three-dimensional reconstruction is carried out.

In micro-image acquisition methods, the demarcation of photographic head adopts the plane reference method of Zhang Zhengyou propositions, with high accuracy And the features such as high robust, applied well in plane reference method；Transmitted to upper computer software due to video camera It is video signal, it is due to before left view and right view are chosen, needing to carry out automatic focusing operation therefore automatic focusing is complete The a certain frame of video signal is used as left view into after, and a certain frame of video signal is regarded as the right side after the anglec of rotation is completed Figure；Using GFTT algorithms characteristic point is carried out extraction be due to the algorithm as feature point detection algorithm under microscopy environment Applied very well, and SIFT algorithms adopt KD-tree search strategys, with higher matching precision and stronger robustness； The method of feature based zone broadening mainly rebuilds the sparse point in space using Feature Points Matching, and by diffusion body surface is being obtained Dense point cloud, the method is simple to operate without the need for any initial information, and arithmetic accuracy depends on the matching of characteristic point.

Specifically, in step S20, the acquisition of left view and right view comprises the following steps process：

S21:Observation micro-image, No. two motors of control and No. five motor work so that micro-image is being regarded Within the scope of open country；

S22：No. three motor work of control, carry out microimaging focusing, until automatic focusing terminates, wherein in focusing During control No. four motors work, in set point, wherein microimaging focusing is adopted at a control number platform inclination angle Focused with the method based on laplacian sharpness evaluation functions；

S23：A two field picture in selecting video signal is stored as left view；

S24：The work of number motor of control room, a two field picture during platform rotational angle β as right view, and Save, wherein 225 ° >=β >=145 °.

In step S22, also include modifying laplacian sharpness evaluation functions, amending method is as follows：

It is by former Laplacian sharpness evaluation functions：

Laplace operator is Second-order Operator, and it is defined as：

Obtained by the definition of second order derived function：

Then Laplacian function representations are：

If it is f (x, y) that space any position p (x, y, z) projects to the pixel on sensor devices, then any position in space Put place p (x, y, z) and be projected as p (x, y) in vertical direction, and the angle between p (x, y, z) and p (x, y) is tilt angle alpha, Therefore the corresponding sharpness evaluation functions of p (x, y) are obtained indirectly for f (x, y) cos θ, the definition of second order derived function is done afterwards as Lower modification：Modification Laplacian functions afterwards can be expressed as：

Finally using amended Laplacian functions as automatic focusing algorithm sharpness evaluation function.

In step s 24,185 ° >=β >=175 ° are chosen.

Specifically, in step s 30, GFTT algorithms are concretely comprised the following steps：

S31：Calculate the second order local derviation (I of each pixel_X)²、(I_y)²、I_xI_y；

S32：Calculate the second-order matrix M of each pixel

In formulaRepresent convolution,

S33：Obtain corresponding two eigenvalue λs of Metzler matrix₁And λ₂；

S34：Choose max (min (λ₁,λ₂)) determine final angle point.

What the present invention was provided realizes that auto-focusing and the microscopic system of three-dimensional reconstruction are realized automatically by image acquiring method Focusing and three-dimensional reconstruction, circuit system and light path design are simple, and integrated level is high；By corresponding upper computer software, it is possible to achieve right The operation of bottom hardware circuit, can thus reduce the error for artificially causing.

Description of the drawings

The above-mentioned and/or additional aspect and advantage of the present invention will become from the description with reference to accompanying drawings below to embodiment It is substantially and easy to understand, wherein：

The microscopic system structural representation for realizing auto-focusing and three-dimensional reconstruction of Fig. 1 present invention；

The microscopic system image acquiring method process schematic of Fig. 2 present invention；

Fig. 3 is that the rotation in the present invention program is taken pictures illustraton of model.

Specific embodiment

It is below in conjunction with the accompanying drawings and concrete real in order to be more clearly understood that the above objects, features and advantages of the present invention Apply mode to be further described in detail the present invention.It should be noted that in the case where not conflicting, the enforcement of the application Feature in example and embodiment can be mutually combined.

Many details are elaborated in the following description in order to fully understand the present invention, but, the present invention may be used also Implemented with being different from mode described here using other, therefore, protection scope of the present invention does not receive following public tool The restriction of body embodiment.

The embodiment of the present invention is further described referring to Fig. 1 to Fig. 3.

As shown in figure 1, the microscopic system of auto-focusing and three-dimensional reconstruction is realized, including：PC, USB2.0 ports it is micro- Photographic head, amplification in front of photographic head are the microlens of 100X, motor, No. two motors, three Number motor, No. four motors, No. five motors and a corresponding step motor drive plate, No. two steps Stepper motor driving plate, No. three step motor drive plates, No. four step motor drive plates, No. five step motor drive plates, single-chip microcomputer, RS232 serial communication modulars.Communicated by RS232 modules between PC and single-chip microcomputer, single-chip microcomputer is according to the PC for receiving Machine (i.e. host computer) command signal to corresponding step motor drive plate sends corresponding control signal, afterwards motor root Corresponding action is performed according to the control signal for receiving, these actions include rotation time, rotation direction and rotating speed.It is concrete to perform Manner of execution is：A number motor is fixedly arranged in the control received on No. 2 platforms and according to a step motor drive plate Signal processed can drive a platform (object stage) to carry out 360 ° of rotations；Its Main Function is to obtain different rotary angle conditions Lower micro-image, and mainly want in the present embodiment by way of rotation come the binocular function in the middle of simulated implementation three-dimensional reconstruction, Therefore when angle rotation is carried out, rotated frequently with the mode plus β angles on the basis of being rotated, wherein 225 ° >=β >=145 ° can realize basic object, and at 185 ° >=β >=175 °, effect is more preferable, be then during control β=180 ° One excellent symmetric position, thus can be with the left view in the middle of simulated implementation binocular three-dimensional reconstruction and the collection work(of right view Energy.The control signal that No. two motors are received according to No. two step motor drive plates can to a platform (object stage), two Number platform and No. three platforms are moved forward and backward；Its Main Function is to adjust micro- object in the middle of the microlens visual field Front and back position；No. three motors can be to a platform (loading according to the control signal that No. three step motor drive plates are received Platform) and No. two platform inclination angle [alpha]s be controlled, precision controlling is in 5 ° >=α >=-5 °, i.e., No. one flat surface and microscope head Between angle be less than or equal to 5 °；Its Main Function is to realize entering the not ipsilateral of micro- object by rotating different angles Row observation, obtains satisfied visual effect, if inclination angle will fog more than 5 ° of micro-images, does not reach and preferably may be used Recognition effect；No. four motors can be to a platform (loading according to the control signal that No. four step motor drive plates are received Platform), No. two platforms and No. three platforms are controlled up and down；Its Main Function is to having adjusted the anglec of rotation and angle of inclination Platform carry out automatic focusing；No. five motors can be to No. one according to the control signal that No. five step motor drive plates are received Platform (object stage), No. two platforms and No. three platforms or so are controlled；Its Main Function is being regarded to adjust micro- object Right position in the middle of open country.Microlens (object lens) are amplified to the micro- object on a platform, and image in photographic head On central sensitive chip, transmitting finally by USB2.0 communication modules carries out image procossing to PC software.Wherein image Process comprises the step of acquisition, image characteristics extraction and matching, the Three-dimensional Gravity of demarcation, left view and the right view of photographic head Build.

In above example, motor can enter line translation and combine with the annexation of each platform, as long as realizing adjustment one The position of platform, for example, can only drive a platform and No. two platform motions to adjust anteroposterior position with No. two motors Put.

The demarcation of video camera：The plane reference method proposed using Zhang Zhengyou is demarcated, of the prior art with reference to text Offer it can be found that the scaling method of video camera there are many kinds, but the plane reference method that Zhang Zhengyou is proposed has high accuracy and high robust The features such as property, applied well in plane reference method.

The acquisition of left view and right view：What is transmitted to PC (host computer) software due to video camera is video signal, because , when left view is carried out and right view chooses, usually a certain frame of selecting video signal is used as left view or right view for this；By In before left view and right view are chosen, need to carry out automatic focusing operation, therefore the backsight for usually completing automatic focusing The a certain frame of frequency signal is used as left view, and the selection of right view, the backsight that usually No. three motor anglecs of rotation are completed The a certain frame of frequency signal is used as right view.

Image characteristics extraction and matching：Image characteristic point is extracted using GFTT algorithms, GFTT algorithm concrete steps For：

1st, the second order local derviation (I of each pixel is calculated_X)²、(I_y)²、I_xI_y。

2nd, the second-order matrix M of each pixel is calculated

WhereinRepresent convolution；

3rd, corresponding two eigenvalue λs of Metzler matrix are obtained₁And λ₂；

4th, max (min (λ are chosen₁,λ₂)) determine final angle point；

List of references finds that feature point detection algorithm is relatively more, and GFTT feature point detection algorithms are obtained under microscopy environment Very well application is given, therefore characteristic point is extracted using GFTT algorithms.

Feature Correspondence Algorithm adopts SIFT algorithms, and characteristic matching SIFT algorithm adopts KD-tree search strategys, with higher Matching precision and stronger robustness.

Three-dimensional reconstruction：

Three-dimensional reconstruction is carried out to micro- object using the method for feature based zone broadening, it is real that micro- object dimensional is rebuild The step of key one of existing three dimensional structure accurate measurement, current Chinese scholars have been developed over various reconstruction techniques.Wherein The more commonly used method having based on silhouettes and the method for feature based zone broadening, are profits based on the method for silhouettes The shape of object is estimated with objects in images profile information, its precision depends primarily on the accuracy of contour of object extraction, obtains The quantity of image sequence and the complexity of object structures are taken, factor of influence is more, and precision is wayward, is confined to convex object Reconstruction；And the method for feature based zone broadening mainly rebuilds the sparse point in space using Feature Points Matching, by spreading To the dense point cloud of body surface, without the need for any initial information, simple to operate, arithmetic accuracy is only dependent upon characteristic point to the method Matching, precision controlling is easily realized therefore three-dimensional reconstruction algorithm is by using the method for feature based zone broadening in the embodiment of the present invention To realize.

In fig. 2, microscopic system image acquiring method process, the method implementation process is as follows：

Start upper computer software first, demarcated by upper computer software video camera, after demarcation is completed, to reaching PC Video image on machine (host computer) software carries out denoising, while micro-image is observed whether within field range, if aobvious Not within field range, upper computer software is by RS232 communication modules to single-chip microcomputer transmitting control commands, control two for micro- image Number motor and No. five motors are rotated, before and after driving a platform to carry out and right position adjustment, to realize microgram As the position in the visual field；When micro-image is in the visual field, and when being in a rational position, upper computer software afterwards By RS232 communication modules to single-chip microcomputer transmitting control commands, No. three motors of control are rotated, and realize a platform and No. two Platform inclination certain angle a；After angle of inclination adjustment is completed, automatic focusing algorithm in the middle of upper computer software, and root are performed By RS232 communication modules to single-chip microcomputer transmitting control commands during the fructufy calculated according to automatic focusing algorithm, No. four are controlled Motor is rotated, and the adjustment of a platform, No. two platforms and No. three platform upper-lower positions is realized, so as to realize automatic focusing； After focusing is completed, the two field picture in selecting video signal stores the image as left view；After storage is completed, on By RS232 communication modules to single-chip microcomputer transmitting control commands, number motor of control is rotated position machine software, realizes that No. one is put down Platform rotates to an angle, and in order to preferably find the image of left view symmetric position, the anglec of rotation is set to 180 °；The anglec of rotation After completing, the two field picture in selecting video signal is as right view and stores the image.Afterwards to the left view to store Image characteristic point extraction is carried out to its characteristic point using GFTT algorithms with right view, afterwards again using SIFT algorithms to extracted Image characteristic point matched；After Feature Points Matching is completed, three-dimensional reconstruction operation is performed on upper computer software；It is three-dimensional After reconstruction is completed, upper computer software respectively to by RS232 communication modules to single-chip microcomputer transmitting control commands, control one, No. two, No. three, No. four, No. five motors rotate and are allowed to platform and return to initial position.

In figure 3, give rotation to take pictures model, next the automatic focusing principle based on rotating model will be situated between Continue.When focusing to micro-image, usually using being focused based on the method for sharpness evaluation function, but using being based on When sharpness evaluation function is focused, micro- plane is all not inclined plane, once because what planar tilt can not be focused on Pixel will increase, although not inclined plane cannot guarantee that all of pixel point focusing in best focus position, but aobvious The general depth of field of speck body is all smaller, therefore the number of out-of-focus pixel can be far smaller than the number of the pixel for focusing on, So when micro- object is in best focus position, the readability of the micro-image integrally seen can receive in human eye Within the scope of.But when micro- planar tilt, it is equivalent to indirectly stretch the depth of field of micro- object, it is clear that work as micro-platform In best focus position when, out of focus pixel will increase.Find in FIG, the object on clinoplain is in sensor devices On projection, have certain difference with projection of the object on sensor devices on horizontal plane, and existing definition evaluates letter The object that number is both on horizontal plane carries out automatic focusing, so if not projecting to shape on sensor devices to clinoplain Pixel into image is done and is processed, then the existing focusing algorithm based on sharpness evaluation function cannot be used, automatically Focusing also just cannot be realized.

List of references finds that conventional sharpness evaluation function has many kinds, and every kind of sharpness evaluation function has respective Pluses and minuses, wherein Laplacian sharpness evaluation functions are：

Laplace operator is Second-order Operator, and it is defined as：

Can be obtained by the definition of second order derived function：

Then Laplacian functions can be expressed as：

If the pixel that space any position p (x, y, z) is projected on sensor devices is f (x, y), then space is any P (x, y, z) is projected as p (x, y) in vertical direction at position, and the angle between p (x, y, z) and p (x, y) is angle of inclination A, therefore the corresponding sharpness evaluation functions of p (x, y) can be indirectly obtained for f (x, y) cos θ；Second order derived function is determined afterwards Justice makes following modification：

Amended Laplacian functions can be expressed as：

Finally using amended Laplacian functions as automatic focusing algorithm sharpness evaluation function；

Method of the algorithm using fixed step size in combination with Step-varied back propagation of wherein focusing is focused.

The ultimate principle and principal character and advantages of the present invention of the present invention, the technology of the industry has been shown and described above Personnel can be that to be mechanically connected can also be electrically connected it should be appreciated that term therein " connection ", " connected " all should broadly understood Connect, can be fixedly connected, integrally connected can also be flexibly connected or be detachably connected, and can be that physical connection can also be Signal connects, and can be that to be joined directly together can also be indirectly connected to by intermediate media, without departing from actual essence connection Relation, the understanding that all should be normally received, these are only the preferred embodiments of the present invention, be not limited to the present invention, For a person skilled in the art, the present invention can have various modifications and variations.It is all the present invention creativeness spirit and Within principle, any modification, equivalent substitution and improvements made etc. should be included within the scope of the present invention.

Claims (8)

1. a kind of microscopic system for realizing auto-focusing and three-dimensional reconstruction, including：Including PC；Microimaging head, with the PC Machine is communicated to connect, and microimaging head front is provided with microlens, and microlens are by the micro- amplification of object to be seen；Monolithic Machine, communicates to connect with the PC and receives the command signal of PC and be converted into control signal output；A number motor, with A number platform connects and for driving a platform to rotate；No. two motors, are connected simultaneously with a platform For driving a platform to move forward and backward；No. three motors, are connected and for driving described one with a platform Number platform inclination angle [alpha]；No. four motors, are connected with a platform and move down on a platform for driving It is dynamic；No. five motors, are connected and for driving a platform to move forward and backward with a platform；A number motor Driving plate, is connected with the single-chip microcomputer and controls the motor work according to the control signal of single-chip microcomputer output；Two Number step motor drive plate, is connected with the single-chip microcomputer and controls No. two electricity according to the control signal of single-chip microcomputer output Machine works；No. three step motor drive plates, are connected and according to the control signal control of single-chip microcomputer output with the single-chip microcomputer No. three motors work；No. four step motor drive plates, are connected and according to the control of single-chip microcomputer output with the single-chip microcomputer Signal processed controls No. four motors work；No. five step motor drive plates, are connected and according to the monolithic with the single-chip microcomputer The control signal of machine output controls No. five motors work.

2. the microscopic system for realizing auto-focusing and three-dimensional reconstruction according to claim 1, it is characterised in that：Also include two Number platform and No. three platforms a, motor is fixedly arranged on No. two platforms and drives a platform to rotate, No. three motors are arranged between No. two platforms and No. three platforms and drive No. two platforms to drive described No. one to put down Platform is inclined, and No. two motors, No. four motors and No. five motors are connected with No. three platforms and drive described No. three to put down Platform drives No. two platforms and a platform to move forward and backward respectively, move up and down and move left and right.

3. the microscopic system for realizing auto-focusing and three-dimensional reconstruction according to claim 1, it is characterised in that：Described No. one Platform inclination angle [alpha] meets 5 ° >=α >=-5 °.

4. micro-image acquisition methods of a kind of microscopic system, realization of the microscopic system as described in claims 1 to 3 is arbitrary The microscopic system of auto-focusing and three-dimensional reconstruction, it is characterised in that the micro-image acquisition methods are comprised the following steps：

S10：The demarcation of photographic head, the plane reference method proposed using Zhang Zhengyou carries out the demarcation of photographic head；

S20：The acquisition of left view and right view, a certain frame of video signal, will used as left view after automatic focusing is completed The anglec of rotation complete after video signal a certain frame as right view；

S30：Image characteristics extraction and matching, are extracted using GFTT algorithms to image characteristic point, using SIFT algorithm conducts The algorithm of Image Feature Matching；

S40：Three-dimensional reconstruction process, using the method for feature based zone broadening three-dimensional reconstruction is carried out.

5. micro-image acquisition methods of microscopic system according to claim 4, it is characterised in that in step S20, it is left The acquisition of view and right view comprises the following steps process：

S21:Observation micro-image, No. two motors of control and No. five motor work so that micro-image is in visual field model Within enclosing；

S22：No. three motor work of control, carry out microimaging focusing, until automatic focusing terminates, wherein in focussing process No. four motor work of middle control, in set point, wherein microimaging focusing adopts base at a control number platform inclination angle Focused in the method for laplacian sharpness evaluation functions；

S23：A two field picture in selecting video signal is stored as left view；

S24：Number motor work of control room a, two field picture during platform rotational angle β is deposited as right view Store, wherein 225 ° >=β >=145 °.

6. micro-image acquisition methods of microscopic system according to claim 5, it is characterised in that in step S22, also Including modifying to laplacian sharpness evaluation functions, amending method is as follows：

Former Laplacian sharpness evaluation functions are：

Laplace operator is Second-order Operator, and it is defined as：

Obtained by the definition of second order derived function：

${\▿}^{2}f(x,y)=f(x+1,y)+f(x-1,y)+f(x,y+1)+f(x,y-1)-4f(x,y)$

Then Laplacian function representations are：

If it is f (x, y) that space any position p (x, y, z) projects to the pixel on sensor devices, then space any position P (x, y, z) is projected as p (x, y) in vertical direction, and the angle between p (x, y, z) and p (x, y) is tilt angle alpha, therefore It is f (x, y) cos θ to obtain the corresponding sharpness evaluation functions of p (x, y) indirectly, the definition of second order derived function is done repair as follows afterwards Change：

Amended Laplacian functions can be expressed as：

$F\left(k\right)=\frac{1}{N*N}\underset{x}{\Σ}\underset{y}{\Σ}\[f(x+1,y)+f(x-1,y)+f(x,y+1)+f(x,y-1))cos\mathrm{\θ}-4f(x,y){\]}^2$

Finally using amended Laplacian functions as automatic focusing algorithm sharpness evaluation function.

7. micro-image acquisition methods of microscopic system according to claim 5, it is characterised in that in step s 24, choosing Take 185 ° >=β >=175 °.

8. micro-image acquisition methods of microscopic system according to claim 4, it is characterised in that in step s 30, GFTT algorithms are concretely comprised the following steps：

S31：Calculate the second order local derviation (I of each pixel_X)²、(I_y)²、I_xI_y；

S32：Calculate the second-order matrix M of each pixel

$M=w(x,y)\⊗\[\begin{array}{cc}{I}_x^2& I_X{I}_Y\\ I_x{I}_y& I_Y^2\end{array}\]$

In formulaRepresent convolution,

S33：Obtain corresponding two eigenvalue λs of Metzler matrix₁And λ₂；

S34：Choose max (min (λ₁,λ₂)) determine final angle point.