CN203164097U - Measure and control device for testing electric rotary biochip and test system - Google Patents

Abstract

The utility model discloses a measure and control device for testing an electric rotary biochip and a test system. The system comprises a biochip test operating platform, an electric rotary drive power supply, a microscope image pickup system consisting of an optical microscopic magnification device and an industrial camera, a measure and control device and an input display device; and the measure and control device is respectively connected with the microscopic image pickup system, the electric rotary drive power supply and the input display device, and the output end of the electric rotary drive power supply is connected with a power connector on the biochip test operating platform. By adopting the scheme, the microscopic image pickup system and the drive power supply are maintained relatively independent, and the special measure and control device is used for substituting the existing PC (personal computer) and a collection card, the measure and control device integrates the function for collecting and processing the image information of the microscopic image pickup system as well as different parameters for controlling the image pickup of the microscopic pickup system and output signal of the drive power supply. The automation of the entire test process of the electric rotary biochip can be realized. The development requirements of the biochip test device such as miniaturization, integration and intelligentization can be met.

Description

A kind of electric rotating biological chip is tested with measure and control device and pilot system

Technical field

The utility model belongs to automatic measure control and biological technical field, relates to the technology of the control that electric rotation test is monitored, specifically a kind of biochip electricity spin stand platform.

Background technology

(electrorotation, ROT) technology is the new technique of a kind of postgraduate's object (as cell) characteristic in the electricity rotation.Its principle is, when the solution that comprises cell or biosome is placed in the specific rotating electric field, cell is because of its dielectric properties difference, and the rotation that produces response is also different, therefore the regularity rotation that cell body can carry various characteristic informations in rotating electric field.By measuring rotary body size, unit rotational speed, following the tracks of rotary body movement locus and individual in conjunction with rule etc., can extract the physical message corresponding with the study subject biochemical character.But this technology produces rotating electric field by the amplitude modulation that applies out of phase at heterogeneous electrode, the AC sine signal of frequency modulation, observes and detect the electric revolving property of biosome under this rotating electric field.This technology be applicable to research cells survival cycle and apoptosis, dynamic monitoring cell physiological state, medicament high flux screening, microorganism separate identify and industrial fermentation with fields such as many biological medicines, cell engineering such as strain improvements.

Electricity rotating biological chip is one of typical application vector of electric rotation technique, be based on the new bio chip of rotating electric field method exploitation, be that the electric rotation test of carrier is the importance that electric rotation technique is used with this chip, application is succeeded in developing and dropped into to the existing multiple electric rotating biological chip of China at present.

But the development of electric rotation technique and the needs that test is used are caught up with in the construction of present electric rotating biological chip test platform far away, are mainly reflected in following several aspect:

1, still there is not perfect robotization test TT﹠C system;

2, test platform integrated, the systematization design is blank both at home and abroad;

3, special-purpose heterogeneous wideband synchro source is too isolated, fails to realize and the centralization of other parts of test platform, unitized management;

4, be unrealized based on PC(or embedded system) image handle the method that Automatic Measurement Technique substitutes existing human eye range estimation.

Building of existing electric rotating biological chip test platform, with heterogeneous wideband synchro source independently as the special-purpose driving power of chip operation, the biological subject volume morphing passes through microscope with the rotation feature, system is input to video monitor through the CCD optical video, again to provide artificial visually examine's method second performance data such as rotational speed.In such platform, system's swelling is loose and is difficult to improve performance; Work such as the initial adjustment of rotating electric field parameter and instant adjusting still need to finish at driving power by hand, have a strong impact on detection efficiency; And the acquisition of the motion feature of tested body, depend on range estimation, not only detection efficiency is low but also cause personal error because of visual fatigue easily.Can't adapt to modern experimental technique development and electricity and revolve the needs that the biochip large-scale promotion is used.

In prior biological chip detection analysis Instrument equipment, collection to the view data of CCD optical video system output, adopt the pattern (Tang Xiaoping of " computing machine+image pick-up card ", Liu Yeyi, .CCD biochip supplies analysis such as Wang Shurong design [J]. micro-nano electronic technology, 2002, (5): 41-44), obtain and to send the further analyzing and processing of PC then to for the view data of analyzing by the scanner sampling earlier.Figure 1 shows that the test macro that is used for electric rotation test in the market, comprise PC, image pick-up card, microscope camera system, four phase wideband synchro sources and electric rotating biological chip on its structure.Four phase wideband synchro sources and microscope camera system are independent separately, the artificial separate operation of palpus, and PC is only as the video image processing terminal.This scenario-frame complexity, bulky, cost is high, the level of resources utilization is low.

In the test macro of existing electric rotation test, processing to view data, be that the regular lattice array still image that utilizes biochip scanner to obtain is handled, separate and handle each sample spot, the parameter that needing to obtain (as area, average gray value, gray integration value and the gray scale maximum of sample point, minimum value etc.).Relatively there are many-sided difference in parameter detecting and other biochip that the cell viability electricity revolves biochip: by the comprehensive detection correlation parameter of dynamic image process information; Dynamic image is derived from the video after the micro-amplification; Be in subjects images such as cell clear, edge fog, the certain background interference of existence inadequately in the solution; Uncertain factor influences image processing etc. in the process of the test.In moving, obtains dynamically in cell that eigenwert such as cell rotational speed need overcome that image is handled and a series of difficult problems such as mark.Handle research at other medical science or biological cell image, mainly concentrate on static cell edges extraction and image and cut apart the aspect.

In addition, adopt the electricity of the mode of computing machine and imaging device combination to revolve the biochip test platform, because its apparatus structure is complicated and problems such as too specialization are operated by system, also restricted the Application of Biochips scope, hindered the biochip industry development (Hu Xiangyu,, Tang Xiaoping, Yan Wei etc.Embedded biochip detects Analytical System Design [J].Computer utility research, 2008,25 (4): 1254-1256).

Summary of the invention

The purpose of this utility model is at above shortcomings in the prior art, proposes a kind of miniaturization, the supporting test apparatus equipment of integrated, intelligentized and electric rotating biological chip.

For achieving the above object, the utility model at first provides a kind of electric rotating biological chip test measure and control device.

It comprises: the driving power control module, be used for to the driving power output control information that produces the four-way power signal that excitation biochip cell rotatablely moves, in order to frequency, the amplitude of each channel power signal and the phase differential between each channel power signal of the power signal that produces of adjusting driving power; Described power signal has the synchronous characteristic of wideband, and its frequency, range value, phase place can arrange;

The microimaging control module is used for the video camera output trigger pip to microscope camera system, starts or stops video acquisition with control;

With the video acquisition processing unit, be used for receiving the video flowing of microscope camera system output, and the video that receives is analyzed, obtain and export the motion state parameters of cell in the tested biochip; Described motion state parameters comprises center displacement, rotational speed, and described analytic process comprises that the image in the video divides that frame obtains, image pre-service, cell edges extraction, celluar localization, cell rotation parameter detect.

Above-mentioned each unit all can adopt pure hardware circuit to make up and forms, also can be by the processor corresponding program instruction of arranging in pairs or groups, and cooperate the communication interface that carries out communication with object-based device to make up again and form.Wherein, the former adopts pure hardware circuit, has deficiencies such as complex structure, integrated level is low, small product size is big, cost is higher.The latter has the realization of being easy to, integrated level height, little, the low cost and other advantages of small product size by for the mode of processor configuration-direct code realizes various controls and processing capacity.So preferred latter of the utility model.

Described driving power control module, microimaging control module and video acquisition processing unit, can be separately configuration processor, storer, communication interface separately, also can share same processor, the staggered corresponding control of corresponding instruction code realization and the processing capacity carried out.In general electric rotating biological Bi sheet experiment, not frequent to the control operation of driving power and microscope camera system, need not accessory resources such as independent configuration processor, storer, so preferred plurality of units shares the scheme of same processor and storer.

Adopt the preferred version of above-mentioned common processor and storer, measure and control device described in the utility model can specifically comprise: processor, be used for stored program instruction code, video data storer, be used for temporary programmed instruction and pending data random access memory, be used for first communication interface and second communication interface of controlled microscope camera system communication and be used for the 3rd communication interface with the communication of controlled electric rotating biological chip drives power supply;

Described first communication interface is connected with the level triggers interface of the video camera of controlled microscope camera system, described processor is exported trigger pip by first communication interface to the level triggers interface of video camera according to programmed instruction, and then the control video camera starts or stops video acquisition.

Described second communication interface is connected with the video output interface of the video camera of controlled microscope camera system, described processor receives the test monitoring video of microscope camera system output by second communication interface, and deposit it in storer, image in the video that described processor divides frame to obtain to receive, after the image pre-service, extract cell edges, centre coordinate in the image, and then calculate motion track and the rotational speed that obtains cell.

Described the 3rd communication interface is connected with the parameter/configuration interface of controlled driving power, and described processor passes through the 3rd communication interface to the frequency of driving power export target power signal, amplitude and the phase place of each channel power signal.

In such scheme, described processor can be various processors with image-capable, as DSP, and PC CPU, ARM etc.Wherein, preferred DSP.Described second communication interface needs the high-speed transfer view data, and first ask not high with the 3rd communication interface mutually to performance, can be various digital I/O interfaces.Accordingly, controlled shooting installation also should have identical or corresponding with it communication interface with driving power.

Existing industrial camera all disposes for the high speed digital interface of output video with for the trigger interface that receives trigger pip.Described high speed digital interface generally adopts USB, 1394, gigabit Ethernet, and is wherein in the majority with USB interface.Therefore, the preferred USB master control of the utility model interface adopts the advantage of this interface to be as second communication interface, need not video camera is transformed, and namely applicable to the existing industrial camera of great majority, versatility is good.Described trigger pip is divided into two kinds of external trigger and instruction triggers.The processor of measure and control device can utilize instruction to trigger video camera by high speed digital interface to start image acquisition, also can trigger the video camera start and stop by producing the mode that a level signal inputs to the video camera trigger interface.

The existing special-purpose driving power that is exclusively used in electric rotating biological chip experiment mainly is a kind of four phase wideband synchro sources, it comprises processing module, high frequency small-signal generation module (DDS), four signal power amplification modules, man-machine interface (keyboard, liquid crystal display etc. are used for manually arranging), host computer interface (being generally the RS232 interface).Wherein DDS can produce four phase power signals, and respectively by four output port outputs, described four signal power amplification module input ends connect four output ports of DDS respectively, and output terminal is four phase output terminal mouths of driving power.Described man-machine interface and host computer interface all link to each other with processing module, described processing module receives the control parameter of man-machine interface or the input of host computer interface, and according to the control parameter that receives the frequency of power signal that DDS produces is set, reach amplitude and the phase place of each phase power signal.

The host computer interface that rotates driving power in view of existing electricity mostly is the RS232 interface, and in order to improve the versatility of measure and control device, the preferred RS232 interface of the utility model is as the 3rd digital interface.Described RS232 interface is connected with the UART interface of described processor by level transferring chip.

Described storer can be read-write memory devices such as the hard disk that is connected with processor, FLASH.

In addition, measure and control device described in the utility model can also comprise mmc card interface, other USB master control interface for other exterior storage of expansion.

Based on above-mentioned measure and control device, the utility model also provides a kind of electric rotating biological chip integrated pilot system.

Microscope camera system, measure and control device, input and display device that this system comprises biochip test operation platform, electricity rotation driving power, is made up of optical microphotograph multiplying arrangement and industrial camera;

Described biochip test operation platform is used for loading electric rotating biological chip, which is provided with the clamping device of electric rotating biological chip and four power connections that link to each other with four phase output terminal mouths of driving power respectively, four power inputs that are loaded in the electric rotating biological chip on the biochip test operation platform are connected with described four power connections respectively;

The objective end of described optical microphotograph multiplying arrangement is over against the electric rotating biological chip that is loaded on the biochip test operation platform, and the pick-up lens of industrial camera links to each other with optical microphotograph multiplying arrangement eyepiece end.

Described measure and control device

Be connected with the trigger interface of industrial camera by first communication interface, the control video camera starts or stops video acquisition.Described measure and control device is connected with the video output interface of the video camera of microscope camera system by second communication interface, receives and handle the experiment video of microscope camera system output, analyzes the motion track and the rotational speed that obtain cell.Described measure and control device is connected with the parameter/configuration interface of driving power by the 3rd communication interface, to the frequency of driving power export target power signal, amplitude and the phase place of each channel power signal.Described measure and control device is connected with input and display device by the four-way communication interface, receives the data that arrange of input and display device input, and motion track and the rotational speed that will analyze the cell of acquisition output to the input and display device demonstration.

Described electricity rotation driving power, comprise processing module, high frequency small-signal generation module (DDS), four signal power amplification modules, the parameter/configuration interface (RS232 that is connected with measure and control device, be connected with measure and control device the 3rd communication interface), can comprise man-machine interface (keyboard, liquid crystal display etc.) in addition, be used for manually driving power directly being arranged.Wherein DDS can produce four phase power signals, and respectively by four output port outputs, described four signal power amplification module input ends connect four output ports of DDS respectively, and output terminal is driving power four phase output terminal mouths.Described man-machine interface and parameter/configuration interface all link to each other with processing module, described processing module receives the configuration parameter of man-machine interface or parameter/configuration interface input, and according to the configuration parameter that receives the frequency of power signal that DDS produces is set, reach amplitude and the phase place of each phase power signal.

In said system, described measure and control device is the core of the integrated pilot system of electric rotating biological chip.Its processor is mainly being born the task of several aspects: 1. the acceptance test device is set the data of input, output test result; 2. control chip test driving power work; 3. receive the video stream from video system; 4. the processing of video information, analysis, quantitative evaluation.Described measure and control device can the aforementioned electric rotation be given birth to chip test measure and control device.

After said system started, processor configuration driven power supply disposed microscope camera system to guarantee to obtain cell electricity image rotating clearly simultaneously to produce corresponding four phase synchronous sine AC signal (power signal); Under the power signal of four phase wideband synchro sources output drove, study subject (cell) rotated in the solution of electric rotating biological chip; Microscope camera system is delivered to dsp system with the trial image (cell rotation diagram) that collects; In DSP, constantly handle acquired image data with specific algorithm, finally draw the cell movement parameter, as the rotational speed of movement locus and the cell of cell barycenter.In the process of system's operation, operating personnel also can upgrade the configuration of driving power and microscope camera system by man machine operation interface.

Scheme of the present utility model keeps the relatively independent of microscope camera system and driving power, substitutes existing PC and capture card with special-purpose measure and control device, the image information function of collection that measure and control device is integrated and processing microscope camera system; And the parameters of control microscope camera system capture and driving power output signal.Realize that electricity revolves biochip and tests whole process automation.Satisfy the miniaturization of biochip testing equipment, integrated, intelligentized growth requirement well.In addition, embedded system cost characteristics low, applying flexible make it possess wide application prospect and market potential in biochip test field.

The communication interface that the utility model is preferably reserved by existing driving power and industrial camera realize measure and control device with theirs alternately.This scheme not only can realize with microscope camera system mutual, to the control of driving power, special-purpose driving power and the application of microscope camera system in cell electricity rotation test TT﹠C system after upgrading more are conducive to upgrade, only need do the corresponding update that just can realize total system of adjusting at system software, greatly reduce complexity and the cycle of exploitation again.

Description of drawings

Fig. 1 is existing electric rotating biological chip test system architecture schematic diagram;

Fig. 2 is the improved electric rotating biological chip test system architecture schematic diagram of the utility model;

Fig. 3 is dsp system structure principle chart described in the utility model;

Fig. 4 is four phase wideband synchro source principle schematic;

Fig. 5 is dsp system and the supporting RS232 interface circuit structural drawing of four phase wideband synchro sources;

Fig. 6 is four phase wideband synchro source guidance panel synoptic diagram;

Fig. 7 is the USB master control interface hardware schematic diagram that dsp system is connected with industrial camera;

Fig. 8 is usb bus enumeration process synoptic diagram;

Fig. 9 is the USB device testing process;

Figure 10 is the utility model pilot system software flow pattern;

Figure 11 is image static treatment process flow diagram;

Figure 12 is cell parameters detection of dynamic process flow diagram;

Figure 13 is gradation of image piecewise linear transform synoptic diagram;

Figure 14 is for connecting the structural element of discontinuous point;

Figure 15 extracts the cell edges process flow diagram for morphological method;

Figure 16 is " annular window " template synoptic diagram;

Figure 17 (a) is template convergent-divergent synoptic diagram;

Figure 17 (b) is the tracking cell process flow diagram;

Figure 18 is that feature point tracking method rotating speed detects synoptic diagram;

Figure 19 is cell movement parameter detecting program flow diagram.

Embodiment

With the form of an example by reference to the accompanying drawings, the integrated pilot system of electric rotating biological chip described in the utility model and measure and control device are described further below.

This example is preferably based on dsp system and designs described measure and control device, and the principle of the integrated pilot system of whole electric rotating biological chip as shown in Figure 2.Wherein four phase wideband synchro sources are as the driving power of electric rotating biological chip test, microscope camera system (being made up of optical microphotograph multiplying arrangement and industrial camera) carries out capture to the electric rotating biological chip in the experiment, and the video data that forms exported to dsp system (measure and control device), described dsp system is controlled the parameter (voltage-phase and conversion frequency) of power signal of four phase wideband synchro sources output and the capture parameter of video camera system (parameter by zoom and focus control capture scope and capture size etc.) simultaneously.

Described dsp system platform is selected the system development platform EBF533 based on the dsp processor ADSP-BF533 of ADI company for use, and its system architecture as shown in Figure 3.

With reference to Fig. 3, described dsp system mainly comprises dsp processor ADSP-BF533, and external unit: memory interface, SDRAM, FLASH, mmc card and hard disk; Man-machine interface, TFT LCD liquid crystal display and touch screen controller; Communication interface, USB device, SPI and RS232 interface.In addition, also comprise logic controller CPLD etc.

ADSP-BF533 is the Blackfin series DSP processor that ADI company produces, and is specially adapted to digital audio/video and image and handles, and has control processing capacity and real time security.Its Specifeca tion speeification is as follows: the highest 600MHz kernel clock frequency, 133MHz peripheral bus frequency on 1200MMACs and the sheet.Two 16 multipliers, two 40 totalizers, two 40 ALUs and four 8 digital video ALUs.32 bit address buses, 4G byte addressing space, 80K byte instruction SRAM, 64K byte data SRAM on the sheet; Can extend out 132M byte external memory storage (ADI.Blackfin Embedded Processor[DB] .http: //www.analog.com/static/imported-files/data sheets/ADSP-BF531 BF532 BF533.pdf.2008/2009).

Above-mentioned dsp system platform to cell electricity rotation test TT﹠C system overall objective to satisfy situation as follows:

(1) memory capacity

As shown in Figure 2, extend out the outer 32M byte SDRAM of sheet on the DSP platform, satisfied in the systematic technical indicator RAM capacity greater than the requirement of 1M byte.Also extend out 4M byte FLASH on this platform, reserved hard-disk interface (IDE), mmc card interface and usb host interface, satisfied the demand of large capacity image data storage fully.

(2) picture size and resolution

TFT LCD liquid crystal display is of a size of 3.5 inches, and (70.08mm * 52.56mm), resolution are 320 * RGB * 240(pixel: 76800), satisfy 2.5 inches, the designing requirement of 76800 pixels.This liquid crystal display links to each other with DSP by parallel Peripheral Interface (PPI), and this interface is supported dma mode.

(3) system interface

The SPI interface, this interface is supported dma mode, interrupts configurable.When the ADSP-BF533 system clock frequency was 100MHz, the SPI transfer rate can reach 25MHz.Touch screen controller TSC2200 links to each other with DSP by the SPI interface.

The RS232 interface, on the DSP platform, identical with the MAX232 function by ADM3202() link to each other with the UART pin of ADSP-BF533.This UART interface is supported dma mode, interrupts configurable.

Most USB 2.0 agreements of supporting of video equipment are on the market changeed the integrated of TT﹠C system for finishing electric rotation test at present, must extend out the usb host interface of supporting USB 2.0 agreements.In addition, also memory stores data such as USB flash disk convenient for users of usb host interface.

Described four phase wideband synchro sources are the power source specials for the rotation test of cell electricity, the synchronous sinusoidal signal of four tunnel equiphase differences of its output is applied on four electrodes of electric rotating biological chip and produces rotating electric field, and the meeting of biological cell under this rotating electric field rotates.Its principle schematic as shown in Figure 4.

In this example, four phase wideband driven in synchronism power technology indexs and the function selected for use are as follows:

Frequency range: in 100Hz ~ 70MHz, frequency is adjustable continuously; But the above single-point of 70MHz arranges 100,120,150 and the 200MHz sine wave.Frequency resolution is 1Hz.

Amplitude output signal: 1 ~ 50V _PpAdjustable.

4 channel signal phase places are freely set for 0 ° ~ 360 °; Every passage is independent or the combination interlock is adjustable, and can select a certain passage is the benchmark passage.

But equipment separate panels operating and setting or by the RS232 interface setting.The setting recording function is all arranged, and parameter can show in liquid crystal display.

Four phase wideband synchro source amplitudes, frequency, phase place is all adjustable, and the RS232 interface of reserving possesses interrupt function, is the integrated facility of having brought of cell electricity rotation test TT﹠C system.

In view of the four phase wideband synchro sources of selecting for use are reserved with the RS232 interface, therefore, this routine described dsp system is selected to be connected with it by the RS232 interface.At the dsp system end, circuit structure as shown in Figure 5 with RS232 interface (the 3rd communication interface) that four phase wideband synchro sources are supporting.Wherein, ADM3202 is the RS232 communication level conversion chip that ADI company produces, and capacitor C is got 0.1uF.The RS232 interface level is followed the EIA-RS-232C standard.

At the dsp system end, in the interface routine of configuration, the serial communication stipulations of four phase wideband synchro sources regulation are as follows:

Baud rate: 9600,1 start bits, 8 data bit, no parity position, a position of rest

Data packet length: 36 bytes

Data packet format: be followed successively by data head+frequency values+cps+standby+amplitude+phase differential, concrete form is as shown in the table:

Data structure according to this communication protocol design is as follows:

For realizing manually the random setting to the work reference of four phase wideband synchro sources, this example also provides four phase wideband synchro source guidance panels that are used for man-machine interaction, as shown in Figure 6.This panel left side be used for the amplitude of display frequency, each passage and phase place and state parameter (as whether working properly, independent or synchronously, output is logical or disconnect); The right side is touch keyboard, and wideband synchro source separate panels is in full accord mutually with four in the button setting.This guidance panel can be that physical surface is analysed, and also can be the picture that shows the tangible operation that forms at touch-screen.

Described video camera system is used for by the optical microphotograph multiplying arrangement electricity being executed the experimentation that changes experimental subjects and carries out imaging, its video camera effect be to sample experimental image of biological cell.Suppose that the cell maximum rotative speed is n _m(r/min), the cell domestic demand that rotates a circle is gathered the N two field picture, and then image acquisition rates must satisfy formula (1):

$c\≥\frac{n_{m}N}{60}\mathrm{fps}---\left(1\right)$

At present, general n in the electric rotation test _m≤ 60r/min, if get N=10, c 〉=10fps then, namely image sampling speed can not be lower than 10fps.

Given this, the video camera of video camera system described in this example is selected USB 2.0 high resolution digital camera MVC1000M for use, and this video camera is mainly used in fields such as micro image analysis processing, picture and text identification.The camera Specifeca tion speeification is as follows:

USB 2.0 interfaces are supported (12Mb/s) transmission at full speed;

Power supply mode: usb bus power supply; The maximum power dissipation of obtaining from usb bus: 128mA;

1/2 inch black and white CMOS photo-sensitive cell of lining by line scan, Pixel Dimensions 5.2 * 5.2 μ m;

Valid pixel (maximum) 1280 * 1024 satisfies 320 * 240 pixel demand (be suitable for observe and handle Minimum requirements);

The requirement of the maximum acquisition rate of 16fps frame per second (1280 * 1024, the time shutter: 10ms), satisfy 10fps(320 * 240);

Output image data form: gray-scale value.In electric rotation test, image is handled based on gray-scale map, adopts the black and white camera can reduce the expense of image acquisition, transmission, storage and processing.

In this example, the interface that described dsp system is connected with microscope camera system is selected the usb host interface for use, its main task is to realize order and the data interaction of dsp system and USB video camera, and then realizes enumerating (identification), disposes the USB video camera and from USB video camera reads image data.

The USB technology is very ripe data transmission technology, the universal standard is the USB2.0 standard at present, at present, the main interface shape of console controller chip that is used for embedded design comprises two kinds: 8 or 16 bit data bus interfaces and other interface shape (SPI, UART etc.), the former is as ISP1161, HL811HS etc., the latter such as MAX3421E, CYP7C67300 etc.Aspect transmission speed, the most the highest support of console controller chip (12Mbps) transmission at full speed that are used for embedded design, also have and support what high speed (480Mbps) was transmitted, as ISP1760, R8A66597, support the usb host controller chip of high-speed transfer be 16 or 32(ISP1760) the bit data bus interface.The dsp system that this example adopts is reserved with the SPI expansion interface, therefore selects for use the USB 2.0 HOST chips that have the SPI interface to expand the usb host interface of dsp system.

MAX3421E and CYP7C67300 are the USB 2.0 console controller chips of SPI interface, all support transmission at full speed.Wherein the SPI interface top speed (bps) of MAX3421E is 26MHz, and the SPI interface top speed of CYP7C67300 is 2MHz.

If image sampling speed is c=10fps, frame-frame time t at interval then _Ff=0.1s, therefore, dsp system must be finished transmission and the processing of a two field picture in 0.1s, if view data is handled the time of reserving 0.02s, then the usb host interface must transmit a frame and is not less than 320 * 240 gray level image (76800 bytes) in 0.08s, and corresponding usb host interface transmission speed is: 76800 * 8/0.08=7.68Mbps.Therefore, the transmission speed of usb host interface is at least 7.68Mbps.Obviously CYP7C67300 does not satisfy the designing requirement of 7.68Mbps; To transmit maximal rate at full speed be 12Mbps to regulation in USB 2.0 agreements, and the SPI speed of MAX3421E then can satisfy the designing requirement of this example and usb protocol, so this example is selected MAX3421E expansion usb host interface for use.

MAX3421E ^[37]Be the SPI interface USB 2.0 host/device controllers that MAXIM company produces, the top speed of its SPI interface reaches 26MHz, sends and accept buffer memory to be two 64 byte IFO.From the usb protocol layer of MAX3421E ^[32]Support, it is supported at full speed and low speed transmissions; Support control transmission, synchronous transmission, interruption transmission and bulk transfer; Self-verifying and conversion DATA0/DATA1 packet; Detect the mistake in all transport-types; Automatic delta frame origin identification (SOF, every millisecond) also guarantees that main frame transmission and frame are initial synchronously; The result of report host requests; The user can realize the setting of data read, device address, end points and transport-type and transmit initialization etc. by the SPI interface; By interruption and transmission object code, the user can understand and respond state and the error of transmission of USB device.In addition, MAX3421E also has 8 road GPIN and 8 road GPOUT, interrupts configurable.The interface shape of MAX3421E and to being supported in of USB 2.0 agreements reduced to a great extent the usb host interface design difficulty, shortened the construction cycle.

Below respectively from two aspects of hardware and software, describing in detail in this example with MAX3421E is the usb host interface of core design.

At first from hardware view, with MAX3421E be the usb host interface principle figure of core design as shown in Figure 7, below respectively to power supply, clock, reset, the design of usb bus, SPI interface, power switch and over-current detection and interrupt circuit is described in detail.

(1) Power supply

The 5V power supply power supply that the usb host expansion board is provided by dsp system, this 5V power supply are the MAX3421E chip power supply by 3.3V voltage adjuster (AMS1117) on the one hand, are the usb bus power supply by USB power switch (TPS2042) on the other hand.

The limit consumed power (maximum) of MAX3421E is 1047mW, AMS1117(3.3V) can provide the 800mA electric current, i.e. the power of 2640mW, and therefore adopting AMS1117 is that the MAX3421E power supply can be satisfied power demand.

As required, usb bus need provide the power of 500mA, and TPS2042 is the dual channel source switch, and under the 5V input, each passage can continue to provide the electric current of 500mA, meets design requirement.

The power supply of usb host expansion board (5V) is provided by dsp system, and usb host expansion board overall power requirement is that 3547mW(1047mW+5V * 500mA), the dsp system power supply is 5V, 2A(10W) Switching Power Supply, can satisfy the demands.

VL is the logic level input pin of MAX3421E among Fig. 7, and link to each other with VCC (3.3V) is as the internal logic datum.

(2) Clock

MAX3421E needs external 12MHz(± 0.25%) crystal oscillator, its load capacitance C _LoadBe the 18pF(maximum), the input capacitance of crystal oscillator input pin XI and output XO pin is 3pF.The value of external capacitor C 1 and C2 (2) calculating in the following manner among Fig. 7:

$C_{\mathrm{Load}}=\frac{\mathrm{<figure-callout\; id="1"\; label="C"\; filenames="HDA00002633173900022.png,HDA00002633173900023.png"\; state="\{\{state\}\}">C</figure-callout>}1\&times;C2}{\mathrm{<figure-callout\; id="1"\; label="C"\; filenames="HDA00002633173900022.png,HDA00002633173900023.png"\; state="\{\{state\}\}">C</figure-callout>}1+\mathrm{<figure-callout\; id="2"\; label="C"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">C</figure-callout>}2}+C_{\mathrm{Stray}}---\left(2\right)$

C wherein _StrayFor stray capacitance (comprising integrated circuit input electric capacity and pcb board electric capacity), be taken as 5pF at this, obtain the C1=C2=26pF(maximum thus), get 22pF during actual design.

The inner USB fixed cycle operator of carrying out of MAX3421E, and after finishing, operation provides interruption, therefore, SPI main frame (DSP) need no timer satisfies the timing requirement of USB, and the register by SPI interface accessing MAX3421E can not impact the actual transmissions process of USB.

(3) Reset

The MAX3421E reset circuit is electrify restoration circuit in this example, and low level is effective.The pulsewidth that resets of MAX3421E is the narrowest to be 200ns, is to guarantee the MAX3421E active homing, and delay time is made as 1ms, gets R0 and C0 and is respectively 1K Ω and 1 μ F.

(4) Usb bus

For preventing bus power source VBUS and signal wire D+ and D-short circuit, the resistance of 33 Ω that near the D+ at USB interface place and D-link, connect respectively.D+ and D-are that the difference mode signal line is right, and as far as possible near wiring, length of arrangement wire and impedance are consistent as far as possible when drawing PCB.

(5) The SPI interface

The SPI interface of dsp system is as main frame (MASTER), and the conduct of the SPI interface of MAX3421E is from equipment (SLAVE), and dsp system is selected input pin by the slave that logic controller produces chip selection signal and MAX3421E

Link to each other, SCLK pin clock is produced by the SPI interface of dsp system.

(6) USB power switch and over-current detection

Adopt TPS2042 as USB power switch and over-current detector in this example.TPS2042 is binary channels over-current detection power switch, and input pin IN is 5V input, and MAX3421E enables pin EN, the switching of gauge tap by certain GPOUT foot control certain passage of TPS2042 processed.When EN was high level, the output OUT of TPS2042 respective channel was 5V.If the usb bus electric current surpasses short circuit thresholding (900mA, representative value) or temperature surpasses 140 ℃, will cut off the electricity supply automatically switch and putting of TPS2042

(Over Current) is low level.

(7) Interrupt

Two interrupt pin INT and GPX are arranged on the usb host expansion board shown in Figure 7, and wherein, INT is the USB service disruption pin of MAX3421E, links to each other with the interrupt logic INT1 of dsp system; GPX is the general interrupts pin (configurable) of GPIN and GPOUT, links to each other with the interrupt logic INT2 of dsp system.When making a mistake in the USB service process, the INT pin of MAX3421E produces look-at-me, and the INT1 pin of dsp system is had no progeny in capturing this and done corresponding response.When bus overload or temperature were too high, certain GPIN pin of MAX3421E was because of by TPS2042's

Pin drags down and triggers the GPX interruption, and then produces look-at-me at the INT2 of dsp system pin.

Secondly from software view, usb host software interarea in this example can be divided into USB system layer and client software layer, below will do detailed introduction to the USB device user program of host controller driver (HCD), USB driver (USBD) and the client software layer of system layer.

Host controller driver(HCD) be the bottom of USB software stack, it is the direct interface of USB driver and console controller (HC) hardware, it realizes HC and abstract with hub Hub are comprised: the data transmission on HC hardware, the HC hardware and distribution and the management of HC resource by a kind of software interface (HCDI) is provided.

This paper is divided into HC read-write and initialization, two parts of data transmission with HCD.

(1) HC read-write and initialization

This part is finished the reading and the configuration of setting and hardware initialization and storage allocation resource etc. of HC register, and mainly function is as shown in the table.

Function	Describe
		byte SPI_RWByte(byteTxData)	The read-write operation of SPI interface
byte H_ReadReg(byteaddr)	Read the value of MAX3421E particular register addr
		void H_WriteReg(byteaddr,byteval)	Put MAX3421E particular register addr value and be val
void Reset_Host(void)	MAX3421E resets
		void MAX3421_Init(void)	The MAX3421E initialization

(2) data transmission

The data transfer request of this section processes USB control transmission and bulk transfer, main function is as shown in the table.

The USB driver(USBD) provide service by two cover mechanism, this two covers mechanism is respectively order mechanism and pipeline mechanism.Usb protocol has been done detailed description to the function that this two covers mechanism will realize, in this article, the major function that the USB driver will be finished is the detection (bus is enumerated) of USB device and the bulk transfer of data.Fig. 8 has described process and the state of Equipment Inspection, and its idiographic flow as shown in Figure 9.

After Equipment Inspection is finished, illustrate that the new equipment that inserts has been ready to have used (the USB client software can be operated equipment).Following table has been listed the main function realization of USB driver herein.

The USB user programEffect is to read the view data that the USB camera acquisition arrives.USB user program function list is as shown in the table.

Function	Describe
		void Read_Image(byte Y，byte X)	Reading a frame pixel is the image of Y * X
void Video_IN(byte Y，byte X，byte F)	Reading in F frame pixel is the image of Y * X

The system software overall flow figure of this routine dsp system as shown in figure 10.

Operating personnel can select following operation by operation interface and touch-screen: four amplitude, frequency and the phase places of each road signal of wideband synchro source mutually are set; Drainage pattern (low noise, normal, highly sensitive), the Exposure mode (automatically, manually) of USB video camera are set; Open the detection of cell movement parameter; Preserve and detect data; Record test video (being stored in external memory storage); Finish test.

In above-mentioned flow process, cell parameters detects and to refer to biological cell experiment video is carried out analyzing and processing with the rotational speed that obtains cell barycenter movement locus and cell etc., it is the core of electric rotation test TT﹠C system software that cell movement parameter detecting algorithm and program thereof realize, be the tracking of final realization cell centroid trajectory and the automatic detection of cell rotational speed, at the cell movement image, the process that this is detected comprises the static treatment of image and two links of detection of dynamic of parameter.The static treatment of image mainly comprise image greyscale transformation, cell edges extraction, cell segmentation, center-of-mass coordinate determine and edge-description that its process flow diagram as shown in figure 11; The parameter detection of dynamic is on the basis of image static treatment, at each independent cell, by barycenter computing, the edge-description to corresponding cell in frame-two field picture, draw angular displacement and the rotational speed of cell barycenter movement locus, calculating cell, its process flow diagram is as shown in figure 12.

Launch successively from the static treatment of cell image and two main aspects of detection of dynamic of cell movement parameter below.

The static treatment of cell image

As shown in figure 11, the static treatment of cell image comprises the description of the definite and cell edges of greyscale transformation, edge extracting, cell segmentation, cell barycenter.Greyscale transformation is the pre-service to image, its objective is in order to obtain cell edges; Cell segmentation is prerequisite to obtain complete cell edges, after correct identification cell, cell to be studied is separated from the image that contains a plurality of cells; The description of barycenter and cell edges is research object with the cell that splits all, and the described function of center-of-mass coordinate and cell edges is the basis of follow-up parameter detection of dynamic.

Greyscale transformation

Owing to reasons such as exposure, environment and noise of equipment, image is being made edge extracting and is being cut apart to go to toward doing original image to strengthen processing, greyscale transformation be one of method of spatial domain figure image intensifying (Li Na. utilize the greyscale transformation method to strengthen digital picture [J]. Beijing Polytechnic College's journal, 2009, (3): 36-37).

Greyscale transformation to as if the image described with gray-scale value of pixel.If coloured image then needs coloured image is converted to gray level image.Basic color space commonly used comprises RGB and YUV etc., wherein the RGB color space is mainly used in colour picture monitor and color video video camera, the YUV color space focuses on gray scale and the aberration of describing image, is easy to compression, transmission and processing, is widely used among the processing of computer video and image.Y in the YUV color space is the gray-scale value of representative image, to be converted to by the coloured image that the RGB color space is described gray level image method (Shen Jun, Ge Suihe. the YUV[J in the computer system]. computer engineering, 2000, (1): 94-9), its available formula (3) is expressed as follows:

Y=0.299R+0.587G+0.114B (3)

Wherein Y represents gray scale (brightness) value of image after the conversion, and R, G, B represent redness, green and the blue component of coloured image respectively.

Greyscale transformation strengthens image with the method for grey scale mapping, such as the contrast that strengthens image.Basic gray-scale transformation method commonly used comprise image inversion, log-transformation, power time conversion, piecewise linear transform etc. (Paul Gonzales. Digital Image Processing [M]. the 2nd edition. Beijing: Electronic Industry Press, 2007), which kind of gray-scale transformation method no matter, its purpose all is for certain a part of characteristic in the outstanding image.At cell image, this paper adopts the method for piecewise linear transform to strengthen the effect of cell edges in the image.The piecewise linear transform synoptic diagram shown in Figure 13 (a), gray scale (Y1, the pixel dynamic range of its gray scale after conversion in Y2) interval is stretched, and its gray scale dynamic range of other interval pixel is compressed; Figure 13 (b) is the special case of piecewise linear transform, is called threshold value again and cuts apart, and namely gray-scale value drops on (Y1, Y2) interval interior pixel, its gray-scale value unification becomes a certain particular value Y2' after conversion, and other pixel then becomes another gray-scale value Y1', and what finally obtain is a width of cloth bianry image.

The functional form that piecewise linear transform shown in Figure 13 and threshold value are cut apart is as follows:

$Z^{\&prime;}=t\left(Z\right)=\left\{\begin{array}{cc}\frac{{\mathrm{<figure-callout\; id="1"\; label="Y"\; filenames="HDA00002633173900022.png,HDA00002633173900023.png"\; state="\{\{state\}\}">Y</figure-callout>}1}^{\&prime;}}{\mathrm{<figure-callout\; id="0"\; label="Y"\; filenames="HDA00002633173900023.png,HDA00002633173900054.png"\; state="\{\{state\}\}">Y</figure-callout>}1}Z,& 0\&le;Z<Y1\\ \frac{{\mathrm{<figure-callout\; id="2"\; label="Y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">Y</figure-callout>}2}^{\&prime;}-{\mathrm{<figure-callout\; id="1"\; label="Y"\; filenames="HDA00002633173900022.png,HDA00002633173900023.png"\; state="\{\{state\}\}">Y</figure-callout>}1}^{\&prime;}}{Y2-Y1}(Z-Y1)+\mathrm{<figure-callout\; id="1"\; label="Y"\; filenames="HDA00002633173900022.png,HDA00002633173900023.png"\; state="\{\{state\}\}">Y</figure-callout>}1,& \mathrm{<figure-callout\; id="1"\; label="Y"\; filenames="HDA00002633173900022.png,HDA00002633173900023.png"\; state="\{\{state\}\}">Y</figure-callout>}1\&le;Z\&le;\mathrm{<figure-callout\; id="2"\; label="Y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">Y</figure-callout>}2\\ \frac{255-{\mathrm{<figure-callout\; id="2"\; label="Y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">Y</figure-callout>}2}^{\&prime;}}{255-Y2}(Z-Y2)+\mathrm{<figure-callout\; id="2"\; label="Y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">Y</figure-callout>}2,& \mathrm{<figure-callout\; id="2"\; label="Y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">Y</figure-callout>}2<Z\&le;255\end{array}\right.---\left(4\right)$

$Z^{\&prime;}=t\left(Z\right)=\left\{\begin{array}{cc}{\mathrm{<figure-callout\; id="2"\; label="Y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">Y</figure-callout>}2}^{\&prime;},& \mathrm{<figure-callout\; id="1"\; label="Y"\; filenames="HDA00002633173900022.png,HDA00002633173900023.png"\; state="\{\{state\}\}">Y</figure-callout>}1\&le;Z\&le;Y2\\ {\mathrm{<figure-callout\; id="1"\; label="Y"\; filenames="HDA00002633173900022.png,HDA00002633173900023.png"\; state="\{\{state\}\}">Y</figure-callout>}1}^{\&prime;},& 0\&le;Z<\mathrm{<figure-callout\; id="1"\; label="Y"\; filenames="HDA00002633173900022.png,HDA00002633173900023.png"\; state="\{\{state\}\}">Y</figure-callout>}1,\mathrm{<figure-callout\; id="2"\; label="Y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">Y</figure-callout>}2<Z\&le;255\end{array}\right.---\left(5\right)$

The method that threshold value shown in this routine employing formula (5) is cut apart is done greyscale transformation to the cell gray level image, obtains the binary map at the outstanding rough edge of cell.

Cell edges is extracted

Edge (border) detects and to belong to the content that image is cut apart in digital image processing field, its objective is the part (another subclass of image) except the scenery edge in the edge (subclass of image) of scenery in the image and the image is separated.It then is method and the sequence of operations of from the image that contains cell the edge separation of cell being come out that cell edges is extracted.

Generally there are following characteristics at the edge of scenery in the image: milder along the variation of edge direction pixel, and change more violent perpendicular to edge direction (normal direction) pixel.Therefore, when to the gray-scale value differentiate of image (replacing differentiate with difference in the digital picture), to obtain extreme value at image border place, many edge detection operators are based on the derivative operation of pixel, as Robert gradient operator, Sobel gradient operator and Laplacian operator etc.Utilize the method for differentiate to obtain main problem of image border existence, that is exactly when having noise in the image, derivative operation can play the effect of amplifying noise undoubtedly, for addressing this problem, smoothing processing is done to image by general elder generation, and then do derivative operation, based on the edge detection method of this thought such as Canny operator etc.The edge detection algorithm that also has other in addition, as template matching method, small echo method of changing with, Hough converter technique and Mathematical Morphology Method etc. (Xia Liangzheng, Li Jiuxian. Digital Image Processing [M]. the 2nd edition. Nanjing: publishing house of Southeast China University, 2005).

Realize the accurate detection of cell rotating speed, its prerequisite is to obtain cell edges accurately, Robert, Sobel and Laplacian operator anti-interference are poor, not good (the Hou Chen of Canny algorithm polarization, Chen Wen. based on the edge extracting [J] of morphology algorithm. forum of the association for science and technology (the second half of the month), 2009, (6): 94-95).Mathematical Morphology Method can be extracted complete edge when suppressing picture noise, and the edge details information that reservation is enriched (Niu Baoqing, Li Wenlin. Edge-Detection Algorithm analysis [J]. Anyang Institute of Technology's journal, 2009, (6): 52-55).This example adopts the method for mathematical morphology to extract cell edges in the image, finally obtains the closed cell edges that single pixel connects.

Next briefly introduce used Mathematical Morphology Method.

Expanding and corroding is the most basic behaviour that bianry image morphology is handled.At bianry image, represent input picture with A, B represents structural element, A, B are two-dimentional integer space Z ²In set.

The inflation table of the A of B is shown:

Its mathematic(al) representation ^[45]For:

This formula represents that the expansion of the A of B is B about the reflection of himself initial point

All displacement z satisfy the set of this formula.

The corrosion of the A of B is expressed as:

Its mathematic(al) representation ^[45]For:

This formula shows that the corrosion of the A of B is the set that B is contained in all displacement z of A.

It is the discontinuous point that connects in the image A that the typical case of expanding uses, and when occurring being interrupted through the image border after the greyscale transformation, available this method is repaired the edge complete.The simplest structural element B that plavini connection discontinuous point adopts as shown in figure 14.

Typical case's application of corrosion is with the uncorrelated details among the selected structural element B elimination bianry image A, reaches the purpose of getting rid of interference, removing noise.

Because the many pixel values of cell interior and cell edges pixel value approach in image, disturb for getting rid of, obtain cell edges accurately, need do to fill to the cell in the image and handle.This operating in is called hole and fills on the Mathematical Morphology Method, it is to set up on the basis of expanding that hole is filled, its objective is the enclosed region that will comprise in the bianry image fill complete, make in this zone pixel value different with background.Hole be filled in cell image feature extraction aspect have the application precedent (Yang Hongwei, Zhang Yun. the blood cell feature extraction of handling based on image [J]. bioinformatics, 2006, (2): 76-78).Therefore, this paper made hole to image and fills processing before extracting cell edges.

In the morphology, the Boundary Extraction of bianry image A can be corroded A by it by choosing suitable structural element B, deducts the result of corroding gained with A then and realizes.If with the border of β (A) expression A, then the mathematic(al) representation of leaching process as the formula (8).

This paper utilizes basic step that Mathematical Morphology Method extracts cell edges in the image as shown in figure 15.

For through the bianry image after the greyscale transformation, select structural element shown in Figure 14 for use.Flow process is as follows: at first carry out expansive working to repair the edge that is interrupted; Then cell is filled complete, for the subsequent extracted cell edges is prepared; Next corrode operation with the incoherent details of filtering, get rid of and disturb; Utilize formula (8) to extract cell edges at last.The cell edges that extracts with the method is the closed cell edges that single pixel connects.

Cutting apart of cell dispersion

The purpose of cell segmentation is to identify each cell and it is separated from the image that contains a plurality of cells, belongs to the object content identified in Digital Image Processing.In the actual tests process some cell can adhere to, phenomenon such as overlapping, deal with more complicated, this paper only limits to the discussion to the cell dispersion partitioning algorithm.

The method of object identification has a variety of in the digital picture, wherein based on the recognition methods of decision theory mainly comprise matching method, best statistical sorter and neural network (Paul Gonzales. Digital Image Processing [M]. the 2nd edition. Beijing: Electronic Industry Press, 2007).In view of the study subject of electric rotation test is a certain fixing colony---cell, the structure of its areal model is relatively easy, therefore adopts matching method to identify cell in this article.

Matching method mainly contains two kinds, and a kind of is minimum distance classifier, and another kind is that this paper selects the latter for use according to relevant matching method.

Before with relevant matches method identification cell, should at first construct a certain cell shuttering.Cell shape is similar to circle or oval, and certain rotational symmetry is arranged, and therefore adopts the variation that can adapt to image in the cell rotary course with a kind of template.According to the general shape of cell, this paper constructs " annular window " template, and its synoptic diagram is shown in Figure 16 (a).

Be initial point with the center of circle, " annular window " template expression formula is as follows:

$h(x,y)=\left\{\begin{array}{cc}1,& d\&le;\sqrt{x^2+{\mathrm{<figure-callout\; id="2"\; label="y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">y</figure-callout>}}^2}\&le;\mathrm{<figure-callout\; id="0"\; label="D"\; filenames="HDA00002633173900023.png,HDA00002633173900054.png"\; state="\{\{state\}\}">\; <figure-callout\; id="0"\; label="D"\; filenames="HDA00002633173900023.png,HDA00002633173900054.png"\; state="\{\{state\}\}">D</figure-callout>\; </figure-callout>}\\ 0,& 0\&le;\sqrt{x^2+{\mathrm{<figure-callout\; id="2"\; label="y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">y</figure-callout>}}^2}<d\end{array}\right.---\left(9\right)$

In the formula, d and D are respectively internal radius and external diameter, if with d _cAnd D _cRepresent cell minimum diameter and maximum gauge respectively, then d and D should satisfy:

{(d,D)|0<d≤d _c<D _c≤D,d,D∈R} (10)

For ease of handling, interior circle that will " annular window " template and cylindrical are respectively with its circumscribed square replacement, and shown in Figure 16 (b), at this moment, formula (9) is adjusted into:

$h(x,y)=\left\{\begin{array}{cc}1,& d\&le;\sqrt{x^2+{\mathrm{<figure-callout\; id="2"\; label="y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">y</figure-callout>}}^2},\left|x\right|\&le;D,\left|y\right|\&le;\mathrm{<figure-callout\; id="0"\; label="D"\; filenames="HDA00002633173900023.png,HDA00002633173900054.png"\; state="\{\{state\}\}">\; <figure-callout\; id="0"\; label="D"\; filenames="HDA00002633173900023.png,HDA00002633173900054.png"\; state="\{\{state\}\}">D</figure-callout>\; </figure-callout>}\\ 0,& 0\&le;\sqrt{x^2+{\mathrm{<figure-callout\; id="2"\; label="y"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">y</figure-callout>}}^2}<d,\left|x\right|\&le;d,\left|y\right|\&le;d\end{array}\right.---\left(11\right)$

TFT LCD liquid crystal display used herein, it is of a size of 70.08mm * 52.56mm, resolution is 320 * 240, average every millimeter 4.57 pixels.Cell dia after the amplification is between 0.5cm ~ 1cm, and its interior ring of the template of then constructing and outer shroud resolution are respectively 23 * 23 and 45 * 45.

Relevant matches is template h _{D * D}With image f _{M * N}(D＜min (M, N)) makes computing cross-correlation, for two dimensional image, Normalized Cross Correlation Function as the formula (12):

$\hat{R}(k,l)=\frac{\underset{i=0}{\overset{M-1}{\mathrm{\&Sigma;}}}\underset{j=0}{\overset{N-1}{\mathrm{\&Sigma;}}}h(i,j)f(i+k,j+l)}{\underset{i=0}{\overset{M-1}{\mathrm{\&Sigma;}}}\underset{j=0}{\overset{N-1}{\mathrm{\&Sigma;}}}{h}^2(i,j)\underset{i=0}{\overset{M-1}{\mathrm{\&Sigma;}}}\underset{j=0}{\overset{N-1}{\mathrm{\&Sigma;}}}{f}^2(i+k,j+l)}---\left(12\right)$

Make computing cross-correlation by " annular window " template and the cell image through edge extracting according to (12), can identify certain independent cell after obtaining its maximum value, the template zone that will comprise the cell that has identified this moment just extracts from original image and cell can be split from the numerous cell of original image, for the description definite and cell edges of follow-up cell center-of-mass coordinate is prepared.

Cell center-of-mass coordinate and edge-description

After being partitioned into each separate cell, with the left upper apex (x of " annular window " housing ₀, y ₀) set up coordinate system as initial point, establish cell edges and constituted by N pixel, on the cell edges arbitrarily the coordinate of any be (x _n, y _n), this pixel value is m _n, cell center-of-mass coordinate (x in this coordinate system then _r, y _r) be:

$x_r=\frac{\underset{n=1}{\overset{N}{\mathrm{\&Sigma;}}}{x}_n{m}_n}{\underset{n=1}{\overset{N}{\mathrm{\&Sigma;}}}{m}_n},$ $y_r=\frac{\underset{n=1}{\overset{N}{\mathrm{\&Sigma;}}}{y}_n{m}_n}{\underset{n=1}{\overset{N}{\mathrm{\&Sigma;}}}{m}_n}---\left(13\right)$

Then can by formula (14) calculate the absolute coordinates of this cell barycenter in original image (x, y).

x=x _r+x ₀,y=y _r+y ₀ (14)

The cell center-of-mass coordinate of obtaining with formula (13) is that initial point is set up polar coordinate system, then (x more arbitrarily on the cell edges _n, y _n) to initial point (x _r, y _r) distance be:

$r_n=\sqrt{{(x_n-x_r)}^2+{(y_n-y_r)}^2}---\left(15\right)$

Itself and pole axis angle are:

${\mathrm{\&theta;}}_n=\arctan \frac{y_n-y_r}{x_n-x_r}---\left(16\right)$

All has ordinal number to (θ _n, r _n) having constituted the edge of cell jointly, cell edges can be represented by formula (17).

r=R(θ) (17)

The detection of dynamic of cell movement parameter

The cell movement parameter detecting is carried out through after the static treatment at cell image, comprises that mainly the description of cell barycenter movement locus and cell rotating speed detect two aspects.

The description of cell barycenter movement locus

Under the normal condition, cell barycenter movement locus can be described with the coordinate sequence (vector) of cell barycenter, and the prerequisite that obtains this sequence or vector is accurately to follow the tracks of subject cell in each two field picture.For the cell dispersion in the electric rotation test, its main forms of motion is rotation, and with translation motion, this translation motion is inviolent in rotation, and moving range is limited in the less space.At this situation, this example proposes a kind of easy method track cells---template convergent-divergent, its basic ideas (shown in Figure 17 (b)) are at first the housing of " annular window " template to be amplified, inside casing dwindles, centered by the absolute center-of-mass coordinate of in last two field picture, being obtained then of cell (as the formula (14)) with corresponding zone in this template " in the circle " present frame, then in this zone, calculate the center-of-mass coordinate of this cell in the present frame with formula (13), use the absolute coordinates of using formula (14) to calculate this cell in the present frame again at last, thereby realize the tracking of cell in the different frame.Template convergent-divergent synoptic diagram is shown in Figure 17 (a), and wherein frame of broken lines is represented original template, realizes the template after frame table shows convergent-divergent.

In successfully following the tracks of each two field picture on the basis of corresponding cell, the coordinate sequence of being made up of this cell barycenter absolute coordinates is the movement locus of cell barycenter.The movement locus of preceding p two field picture cell barycenter is shown in (186).

P(f)={(x,y) ₁,(x,y) ₂,…,(x,y) _p} (18)

If regular the following of cell barycenter movement locus can obtain the equation of cell barycenter movement locus by the mode of curve match.

The cell rotating speed detects

The cell rotational speed is the angular velocity of cell, and the prerequisite of angular velocity detection is to detect the angular displacement of cell.Two kinds of cell angular velocity detection algorithms are arranged: feature point tracking method and cross-correlation method in the prior art.

Described feature point tracking ratio juris is: look for a unique point in cell edges, follow the tracks of this characteristic point coordinates in each two field picture, utilize the change calculations of this unique point coordinate between frame-two field picture to go out the angle that cell turns over.

When using this method, can choose the corresponding marginal point of cell greatest radius (characteristic radius) as unique point, namely on the cell edges apart from cell barycenter point farthest.Be that initial point is set up rectangular coordinate system with the cell barycenter, the unique point coordinate of establishing cell to be studied in the p two field picture is (x _p, y _p), its expression formula is:

$\left\{(x_p,y_p)\right|\max \left(\sqrt{{\mathrm{<figure-callout\; id="2"\; label="x\; i"\; filenames="HDA00002633173900021.png,HDA00002633173900022.png"\; state="\{\{state\}\}">x</figure-callout>}}_i^{}+y_i^2}\right)\}---\left(19\right)$

(x wherein _i, y _i) be any coordinate of any on the cell edges.The angle that cell turns between p frame and the reference frame (b frame) is:

${\mathrm{\&Delta;\&theta;}}_p={\mathrm{\&theta;}}_p-{\mathrm{\&theta;}}_b=\arctan \frac{y_p}{x_p}-\arctan \frac{y_b}{x_b}---\left(20\right)$

If the time difference is Δ t between frame-frame, then can be drawn by formula (21) to p frame cell mean angular velocity from the b frame.

${\stackrel{\&OverBar;}{\mathrm{\&omega;}}}_p=\frac{\underset{i=b}{\overset{p-1}{\mathrm{\&Sigma;}}}{\mathrm{\&Delta;\&theta;}}_i}{(p-b)\mathrm{\&Delta;t}}---\left(21\right)$

Levy a synoptic diagram of tracing detection cell rotating speed as shown in figure 18.

Described cross-correlation method detects in distance, the speed context of detection has more application, and for example the mode of handling with image utilizes cross-correlation method to detect particle rotating speed [49].Cross-correlation method operation to as if the described cell edges of different frame Chinese style (17).

For the one dimension function, the cross correlation function discrete expression is:

${\hat{R}}_{\mathrm{xy}}\left(k\right)=\frac{1}{N}\underset{n=0}{\overset{N-1}{\mathrm{\&Sigma;}}}y\left(n\right)x(n-k)---\left(22\right)$

The cross correlation function that calculates p frame and reference frame (b frame) r according to formula (22) is as follows:

Wherein

Be deviation angle.Obtain in the formula (23)

Maximal value corresponding

Can draw the angle that cell turns between two frames:

Can be drawn the angular velocity of cell at last by formula (21).

Comprehensive above the description, cell movement parameter detecting program flow diagram of the present utility model as shown in figure 19.

After receiving " cell parameters detection " order, just begin to detect the cell movement parameter according to flow process, after receiving " stopping to detect " order, stop.Test findings and some variable parameters are temporary among the RAM, and the user can order in the memory storages such as these data being saved in Flash by " detecting the data storage ".

Above algorithm realizes with MATLAB on PC that earlier empirical tests effectively back (detailed content please refer to chapter 5 algorithm test portion) is transplanted in dsp system.The utility model has mainly been taked Several Measures in the program portable process:

(1) consideration of global variable and local variable

Global variable does not belong to some functions, but belongs to a source files of program, and its action scope is whole source program.When variable was declared as local static variable, it can only use in the local action territory, go beyond the scope and can not use, but it is but taking internal memory all the time.So in the compiling procedure of code, should adopt global variable to reduce the expense of the re-defining of like variable, economize on hardware resource as far as possible.

(2) setting of data type

Data type in the MATLAB computation process adopts the double-precision floating point type usually, and its dynamic range is big, generally can not consider the overflow problem of variable.The ADSP-BF533 processor is fixed-point processor, in data handling procedure, should avoid using the floating type data as far as possible, each variable, particularly intermediate variable should fully be estimated its dynamic range when the definition data type, cause makeing mistakes of result of calculation to avoid overflowing of factor certificate.The code of this paper algorithm is before transplanting to dsp system, the dynamic range of its variable obtains monitoring in the PC debugging of algorithm, so when the DSP of algorithm realizes, defined data type rightly, there is not the data overflow problem, not only guaranteed result's accuracy but also saved the expense of system resource.

(3) use of C language library function

The C language has abundant built-in function, can consider to call the built-in function of C language in the implementation procedure of algorithm, realizes the construction cycle of difficulty, shortening software with the code that reduces algorithm.

(4) use of division arithmetic

DSP has very strong multiply-add operation ability, in the process that code is write, based on multiplication and addition, adopts division arithmetic less as far as possible.

(5) processing of redundant circulation

In the code implementation procedure of the algorithm of Digital Image Processing a lot of cyclic processes are arranged, adopt rational algorithm and program structure can effectively reduce the cyclic redundancy amount, improve program operation speed.

Claims (6)

1. measure and control device is used in an electric rotating biological chip test, it is characterized in that, comprising:

The driving power control module, be used for to the driving power output control information that produces the four-way power signal that excitation biochip cell rotatablely moves, in order to frequency, the amplitude of each channel power signal and the phase differential between each channel power signal of the power signal that produces of adjusting driving power;

The microimaging control module is used for the video camera output trigger pip to microscope camera system, starts or stops video acquisition with control;

With the video acquisition processing unit, be used for receiving the video flowing of microscope camera system output, and the video that receives is analyzed, obtain and export the motion state parameters of cell in the tested biochip.

2. electric rotating biological chip according to claim 1 is tested and is used measure and control device, it is characterized in that, this device specifically comprises processor, is used for the stored program instruction code, the storer of video data, be used for temporary programmed instruction and pending data random access memory, be used for first communication interface and second communication interface of controlled microscope camera system communication and be used for the 3rd communication interface with the communication of controlled electric rotating biological chip drives power supply;

Described first communication interface is connected with the level triggers interface of the video camera of controlled microscope camera system, described processor is exported trigger pip by first communication interface to the level triggers interface of video camera according to programmed instruction, and then the control video camera starts or stops video acquisition;

Described second communication interface is connected with the video output interface of the video camera of controlled microscope camera system, described processor receives the test monitoring video of microscope camera system output by second communication interface, and deposit it in storer, image in the video that described processor divides frame to obtain to receive, after the image pre-service, extract cell edges, centre coordinate in the image, and then calculate motion track and the rotational speed that obtains cell;

Described the 3rd communication interface is connected with the parameter/configuration interface of controlled driving power, and described processor passes through the 3rd communication interface to the frequency of driving power export target power signal, amplitude and the phase place of each channel power signal.

3. electric rotating biological chip according to claim 2 is tested and is used measure and control device, it is characterized in that, described processor is dsp processor, described second communication interface is USB master control interface, described the 3rd communication interface is the RS232 interface, and described RS232 interface is connected with the UART interface of described processor by level transferring chip.

4. integrated pilot system of electric rotating biological chip based on the described measure and control device of claim 1, it is characterized in that, microscope camera system, the measure and control device that this system comprises biochip test operation platform, electricity rotation driving power, is made up of optical microphotograph multiplying arrangement and industrial camera, reach input and display device;

Described biochip test operation platform is used for loading electric rotating biological chip, which is provided with the clamping device of electric rotating biological chip and four power connections that link to each other with four phase output terminal mouths of driving power respectively, four power inputs that are loaded in the electric rotating biological chip on the biochip test operation platform are connected with described four power connections respectively;

The objective end of described optical microphotograph multiplying arrangement is over against the electric rotating biological chip that is loaded on the biochip test operation platform, and the pick-up lens of industrial camera links to each other with optical microphotograph multiplying arrangement eyepiece end;

Described measure and control device is connected with the trigger interface of video camera by first communication interface, and the control video camera starts or stops video acquisition; Described measure and control device is connected with the video output interface of the video camera of microscope camera system by second communication interface, receives and handle the experiment video of microscope camera system output, analyzes the motion track and the rotational speed that obtain cell; Described measure and control device is connected with the parameter/configuration interface of driving power by the 3rd communication interface, to the frequency of driving power export target power signal, amplitude and the phase place of each channel power signal; Described measure and control device is connected with input and display device by the four-way communication interface, receives the data that arrange of input and display device input, and motion track and the rotational speed that will analyze the cell of acquisition output to the input and display device demonstration.

5. the integrated pilot system of electric rotating biological chip according to claim 4, it is characterized in that, described electricity rotation driving power comprises processing module, high frequency small-signal generation module, four signal power amplification modules, the parameter/configuration interface that is connected with measure and control device; Described high frequency small-signal generation module can produce four phase power signals, and respectively by four output port outputs, described four signal power amplification module input ends connect four output ports of high frequency small-signal generation module respectively, and output terminal is driving power four phase output terminal mouths; Described parameter/configuration interface links to each other with processing module, described processing module receives the configuration parameter of parameter/configuration interface input, and according to the configuration parameter that receives the frequency of high frequency small-signal power signal that generation module produces is set, reach amplitude and the phase place of each phase power signal.

6. the integrated pilot system of electric rotating biological chip according to claim 5 is characterized in that, described electricity rotation driving power also comprises the Man Machine Interface that driving power is directly arranged for manually.

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