CN105403989B - Nematode identifying system and nematode recognition methods - Google Patents

Nematode identifying system and nematode recognition methods Download PDF

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Publication number
CN105403989B
CN105403989B CN201510713008.7A CN201510713008A CN105403989B CN 105403989 B CN105403989 B CN 105403989B CN 201510713008 A CN201510713008 A CN 201510713008A CN 105403989 B CN105403989 B CN 105403989B
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axis
nematode
axis guide
image information
objective table
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CN105403989A (en
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徐葳
李越
曲嘉楠
王永才
宋磊
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Tsinghua University
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Tsinghua University
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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/365—Control or image processing arrangements for digital or video microscopes
    • G02B21/367—Control or image processing arrangements for digital or video microscopes providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T2207/00—Indexing scheme for image analysis or image enhancement
    • G06T2207/10—Image acquisition modality
    • G06T2207/10056—Microscopic image

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  • Engineering & Computer Science (AREA)
  • Multimedia (AREA)
  • Physics & Mathematics (AREA)
  • Computer Vision & Pattern Recognition (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Image Processing (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

The present invention, which provides a kind of nematode identifying system and nematode recognition methods, the nematode recognition methods, to be included：The culture of nematodes ware that culture has nematode is configured at objective table；The nematode and obtaining observed by digit microscope in nematode culture dish can meet the nematode image information of image recognition requirement；Image recognition is carried out by control terminal-pair nematode image information, including：Gray proces are carried out to nematode image information；Nematode image information after gray proces is smoothed；And self-adaption thresholding processing and contours extract are carried out to the nematode image information after smoothing processing, to identify the nematode in nematode image information.Compared to prior art, the present invention has identification method simple and quick, and nematode discrimination is high and versatility is good easy to utilize, can be identified in having the advantages that miscellaneous bacteria and the even environment of uneven illumination.

Description

Nematode identifying system and nematode recognition methods
Technical field
The invention belongs to biomedical sector, especially, is related to a kind of nematode identifying system and nematode recognition methods.
Background technology
Caenorhabditis elegans (Caenorhabditis elegans) is that a kind of nontoxic, non-parasitism (can independently give birth to Deposit) nematode, its adult length is about one millimeter, the whole body transparent, very thin body.Full-length genome is completed as first to survey The multicellular organism of sequence, Caenorhabditis elegans (hereinafter referred to as nematode) is widely used as model organism in biological study to be made With.
Nematode also implies that the demand that this operation is identified for nematode for the meaning of biological study, many at present real Test room and still carry out manual identified by by digit microscope, on the one hand manual identified nematode expends energy very much, on the other hand Efficiency is not also high.
Further, those skilled in the art know Digital Image Processing and mode identification technology applied to nematode automatically Other field.But existing nematode image automatic identification technology discrimination is not high, poor robustness, majority be difficult to push to it is actual should With.
The content of the invention
It is an object of the invention to provide a kind of nematode identifying system and nematode recognition methods, to solve prior art center line Recognition methods existing for worm identification technology is cumbersome, wastes time and energy and the problems such as discrimination is relatively low.
The present invention provides a kind of nematode identifying system on the one hand, including：Frame；In the frame, for configuring The objective table of culture of nematodes ware；Digit microscope in the frame and above the objective table, for observing State the nematode in culture of nematodes ware and obtain the nematode image information that can meet image recognition requirement；And control terminal, with institute Digit microscope communication connection is stated, next nematode image information is transmitted at the digit microscope and to the line for receiving Worm image information carries out image recognition；The control terminal includes：Gray proces unit, for entering to the nematode image information Row gray proces；Smoothing processing unit, it is connected with the gray proces unit, for carrying out gray scale to the gray proces unit The nematode image information after processing is smoothed；Self-adaption thresholding processing unit, with the smoothing processing unit Connection, carried out for the nematode image information after being smoothed to the smoothing processing unit at self-adaption thresholding Reason；Contours extract unit, it is connected with the self-adaption thresholding processing unit, for the self-adaption thresholding processing unit Carry out the nematode image information after self-adaption thresholding processing and carry out contours extract, to identify the nematode image information In nematode.
In one embodiment of the present invention, the nematode identifying system also includes the observation of the corresponding culture of nematodes ware Light source, the observation light source are to provide the uniform parallel towards the culture of nematodes ware positioned at the bottom of the culture of nematodes ware Light source.
In one embodiment of the present invention, the uniform parallel light source includes LED multi-point sources, the LED multiple spots light Source is placed on the objective table, and a frosted glass is configured with the LED multi-point sources, and the culture of nematodes ware is placed in described On frosted glass.
In one embodiment of the present invention, the nematode identifying system also includes：In the frame, for adjusting The three-dimensional motion mechanism of the digit microscope and the objective table relative position relation；And with the three-dimensional motion mechanism and The control panel of the control terminal connection, described three are controlled for receiving the control instruction from the mobile terminal of communication connection Tie up motion.
In one embodiment of the present invention, the three-dimensional motion mechanism includes：The digit microscope is driven to be transported along X-axis Dynamic X-axis telecontrol equipment；The Y-axis telecontrol equipment for driving the digit microscope to be moved along Y-axis；And the driving objective table edge The Z axis telecontrol equipment of Z axis motion.
In one embodiment of the present invention, the X-axis telecontrol equipment includes：X-axis guide rod, X-axis screw mandrel, X-axis are oriented to Block and X-axis motor, the X-axis guide rod and the X-axis screw mandrel are set parallel along X-axis, and the digit microscope is fixed on The X-axis guide pad and the X-axis guide pad are sheathed on the X-axis guide rod, and the X-axis motor set is located at the X-axis screw mandrel And it is connected with the X-axis guide pad；The Y-axis telecontrol equipment includes：Y-axis guide rod, Y-axis screw mandrel, Y-axis guide pad and Y-axis Motor；Y-axis guide rod is two, and the opposite sides at the top of the frame is respectively in along Y direction；The Y-axis screw mandrel with One Y-axis guide rod is set parallel；Y-axis guide pad is two, is sheathed on two Y-axis guide rods respectively, and, it is described The opposite end of X-axis guide rod and the X-axis screw mandrel is inserted in two Y-axis guide pads respectively；The y-axis motor is sheathed on One Y-axis screw mandrel and it is connected with a Y-axis guide pad；And the Z axis telecontrol equipment includes：Z axis guide rod, Z Axial filament bar and Z axis motor, the Z axis guide rod and the Z axis screw mandrel are set parallel along Z axis, and the objective table is sheathed on The Z axis guide rod and the Z axis screw mandrel, the Z axis motor set are connected located at the Z axis screw mandrel and with the objective table.
In one embodiment of the present invention, the X-axis telecontrol equipment includes：X-axis guide rod, X-axis synchrome conveying belt, X Axle guide pad and X-axis motor, the X-axis guide rod and the X-axis synchrome conveying belt are set parallel along X-axis, the number Microscope is fixed on the X-axis guide pad and the X-axis guide pad is sheathed on the X-axis guide rod, and the X-axis motor set is located at The X-axis synchrome conveying belt and it is connected with the X-axis guide pad；The Y-axis telecontrol equipment includes：Y-axis guide rod, Y-axis are synchronous Conveyer belt, Y-axis guide pad and y-axis motor；Y-axis guide rod is two, and the top of the frame is respectively in along Y direction Opposite sides；The Y-axis synchrome conveying belt is set parallel with a Y-axis guide rod；Y-axis guide pad is two, respectively Two Y-axis guide rods are sheathed on, and, the opposite end of the X-axis guide rod is inserted in two Y-axis and is oriented to respectively Block；The y-axis motor is sheathed on a Y-axis synchrome conveying belt and is connected with a Y-axis guide pad；And the Z Axle telecontrol equipment includes：Z axis guide rod, Z axis screw mandrel and Z axis motor, the Z axis guide rod and the Z axis screw mandrel are along Z Axle is set parallel, and the objective table is sheathed on the Z axis guide rod and the Z axis screw mandrel, and the Z axis motor set is located at the Z Axial filament bar and it is connected with the objective table.
In one embodiment of the present invention, the digit microscope be fixed on by a microscope mounting bracket it is described X-axis guide pad.
In one embodiment of the present invention, the objective table is rectangular mesa or triangle table top, the one of the objective table Individual side is as the connection side being connected with the Z axis telecontrol equipment；And in the Z axis telecontrol equipment, including two institutes Z axis guide rod and a Z axis screw mandrel are stated, the Z axis screw mandrel is placed in the middle, and two Z axis guide rods are located at the Z axis screw mandrel Opposite sides.
In one embodiment of the present invention, the X-axis motor, the y-axis motor and the Z axis motor are step Stepper motor.
Nematode identifying system provided by the invention, including organic frame, objective table, digit microscope and control terminal, institute Control terminal is stated to transmit the nematode image information come at the digit microscope and carry out image to the nematode image information Identification, to identify the nematode in the nematode image information.Compared to prior art, nematode identifying system of the present invention, which has, to be known Other mode is simple and quick, and nematode discrimination is high and versatility is good easy to utilize, can have miscellaneous bacteria and uneven illumination even Environment in the advantages that being identified.
The present invention provides a kind of nematode recognition methods on the other hand, including：The culture of nematodes ware that culture has nematode is matched somebody with somebody It is placed in objective table；Nematode in the culture of nematodes ware as described in digital micrograph sem observation simultaneously obtains the line that can meet image recognition requirement Worm image information；And the nematode image information is received by control terminal and image knowledge is carried out to the nematode image information Not, including：Gray proces are carried out to the nematode image information；The nematode image information after gray proces is carried out smooth Processing；And self-adaption thresholding processing and contours extract are carried out to the nematode image information after smoothing processing, with identification The nematode gone out in the nematode image information.
In one embodiment of the present invention, the culture of nematodes ware that culture has nematode is configured at objective table, including：Will training The culture of nematodes ware that supporting has nematode is positioned on objective table；And provide observation light source for the culture of nematodes ware.
In one embodiment of the present invention, the observation light source is positioned at the bottom of the culture of nematodes ware, to provide Towards the uniform parallel light source of the culture of nematodes ware.
In one embodiment of the present invention, the nematode recognition methods also includes carrying out leveling to the objective table.
In one embodiment of the present invention, nematode in the culture of nematodes ware as described in digital micrograph sem observation simultaneously obtains energy Meet the nematode image information of image recognition requirement, including：By digit microscope obtain observe on the objective table The image information of nematode in the upper culture of nematodes ware, described image information transmission is communicated to connect to the digit microscope Control terminal；By it is described control terminal according to described image information and output control instruct and three-dimensional is controlled by control panel Motion is to adjust the relative position of the digit microscope and the objective table, so as to realize auto-focusing；And pass through The digit microscope obtains the nematode image information that can meet image recognition requirement.
In one embodiment of the present invention, three-dimensional motion mechanism is controlled to adjust the digit microscope and the loading The relative position of platform, including：Adjust motion of the digit microscope in the plane that X-axis and Y-axis are formed；And adjustment institute State motion of the objective table on Z axis.
In one embodiment of the present invention, three-dimensional motion mechanism is controlled to adjust the digit microscope and the loading The relative position of platform, including：Adjust three-dimensional motion mechanism of the digit microscope in X-axis, Y-axis and Z axis.
In one embodiment of the present invention, the smoothing processing uses medium filtering.
In one embodiment of the present invention, self-adaption thresholding is carried out to the nematode image information after smoothing processing Processing and contours extract, including：
Centered on each pixel, the pixel in around it one square area delimited is averaged to determine this Adaptive threshold at a little；
The pixel placement that will be above the adaptive threshold is 0, and the pixel placement that will be less than the adaptive threshold is 255；
After the result of adaptive threshold is obtained, all white areas are found by extracting in image outmost outline The pixel on the border in domain, calculate the area that each outline is included and find out the profile that those areas exceed predefined size, delete Fall be not nematode the white portion no more than predefined size, using area exceed predefined size white portion as target area Domain, rectangular bounding box and the circumscribed circle of these target areas are sketched out to mark their position in the picture.
Nematode recognition methods provided by the invention, by the nematode in culture of nematodes ware described in digital micrograph sem observation and is obtained The nematode image information of image recognition requirement can be met by taking, to the nematode image information carry out gray proces, smoothing processing, from After adapting to thresholding processing and contours extract, the nematode in the nematode image information is identified.Compared to prior art, Nematode recognition methods of the present invention has identification method simple and quickly, nematode discrimination is high and versatility is good easy to utilize, It can be identified in having the advantages that miscellaneous bacteria and the even environment of uneven illumination.
Brief description of the drawings
Fig. 1 is the overall structure diagram in one embodiment of nematode identifying system of the present invention.
Fig. 2 is the schematic diagram for putting culture of nematodes ware in Fig. 1 on objective table.
Fig. 3 is for the structural representation for the microscope mounting bracket that digit microscope is fixed on X-axis guide pad in Fig. 1 Figure.
Fig. 4 is the functional block diagram that terminal is controlled in nematode identifying system of the present invention.
Fig. 5 is the nematode image schematic diagram after being handled using self-adaption thresholding.
Fig. 6 is using the nematode image schematic diagram after contours extract.
Fig. 7 is the schematic flow sheet of nematode recognition methods of the present invention in one embodiment.
Fig. 8 is the schematic flow sheet of step S57 refinement step in Fig. 7.
Component label instructions
101 frames
102 luggage carriers
103 objective tables
104 bases
105 digit microscopes
106 observation panels
107 microscope mounting brackets
108 spiders
111 X-axis guide rods
113 X-axis synchrome conveying belts
115 X-axis guide pads
119 Y-axis guide rods
121 Y-axis guide pads
127 Z axis guide rods
129 Z axis screw mandrels
20 control terminals
201 gray proces units
203 smoothing processing units
205 self-adaption thresholding processing units
207 contours extract units
30 culture of nematodes wares
S51~S57 steps
S571~S577 steps
Embodiment
Illustrate embodiments of the present invention below by way of specific instantiation, those skilled in the art can be by this specification Disclosed content understands other advantages and effect of the present invention easily.The present invention can also pass through specific realities different in addition The mode of applying is embodied or practiced, the various details in this specification can also be based on different viewpoints with application, without departing from Various modifications or alterations are carried out under the spirit of the present invention.It should be noted that in the case where not conflicting, following examples and implementation Feature in example can be mutually combined.
It should be noted that the schema provided in following examples only illustrates the basic structure of the present invention in a schematic way Think, the component relevant with the present invention is only shown in schema rather than is painted according to component count, shape and the size during actual implement System, kenel, quantity and the ratio of each component can be a kind of random change during its actual implementation, and its assembly layout kenel also may be used Can be increasingly complex.
Fig. 1 is referred to Fig. 3, it is shown that the structural representation of nematode identifying system of the present invention in one embodiment, Wherein, Fig. 1 is the overall structure diagram of nematode identifying system of the present invention, and Fig. 2 is to put culture of nematodes ware in Fig. 1 on objective table Schematic diagram, Fig. 3 is the structural representation of microscope mounting bracket for being used to digit microscope being fixed on X-axis guide pad in Fig. 1 Figure.
It should be noted that the nematode identified needed for nematode identifying system of the present invention is with Caenorhabditis elegans Illustrated exemplified by (Caenorhabditis elegans), Caenorhabditis elegans be it is a kind of it is nontoxic, non-it is parasitic (can be with Independently survive) nematode, its adult length is about one millimeter, the whole body transparent, very thin body.Full base is completed as first Because of a group multicellular organism for sequencing, Caenorhabditis elegans (hereinafter referred to as nematode) is widely used as pattern life in biological study Thing uses.
As shown in figure 1, nematode identifying system of the present invention, including：Frame 101, objective table 103, digit microscope 105, three Tie up motion, control panel and control terminal 20.
Above-mentioned each component is described in detail below.
Frame 101 be rectangular-shape (also including cubic) outer frame skeleton structure, inner hollow, wherein, frame 101 top frame and several side frames offer opening, are easy to observe and take article.
Objective table 103 is in the frame, for configuring nematode culture dish 30.In a preferred embodiment, nematode is trained Supporting ware 30 can be positioned on objective table 103 by certain mode or instrument.Culture has nematode in culture of nematodes ware 30, in reality In, nematode is stored in air-conditioned room, and temperature is maintained between 20 DEG C to 25 DEG C, in case nematode influences into stress situation Experiment.In addition, it need to preferably be preserved for the culture medium for cultivating nematode in 4 DEG C of environment.
Digit microscope 105, can by digit microscope 105 in frame 101 and positioned at the top of objective table 103 Clearly observe nematode in culture of nematodes ware 30 and obtain the nematode image information of the corresponding nematode.
Three-dimensional motion mechanism is mainly used for adjusting digit microscope 105 and the relative position relation of objective table 103, so that The image information of nematode in nematode culture dish 30 on objective table 103 can more clearly be obtained in favor of sight by obtaining digit microscope 105 Examine and identify.In a preferred embodiment, the three-dimensional motion mechanism includes：For driving digit microscope 105 along X-axis The X-axis telecontrol equipment of motion；For the Y-axis telecontrol equipment for driving digit microscope 105 to be moved along Y-axis；And carried for driving The Z axis telecontrol equipment that thing platform 103 moves along Z axis.And in optional other embodiment, the three-dimensional motion mechanism can also wrap Include：For the X-axis telecontrol equipment for driving digit microscope 105 to be moved along X-axis；For driving digit microscope 105 to be moved along Y-axis Y-axis telecontrol equipment；And the Z axis telecontrol equipment for driving digit microscope 105 to be moved along Z axis.
For above-mentioned preferred embodiment, in the first embodiment, the X-axis telecontrol equipment includes：X-axis guide rod 111, X-axis synchrome conveying belt 113, X-axis guide pad 115 and X-axis motor (not identifying in the drawings), X-axis guide rod 111 and X-axis are same Step conveyer belt 113 is set parallel along X-axis, and X-axis guide pad 115 is sheathed on the X-axis guide rod, and digit microscope 105 is fixed In on X-axis guide pad 115, X-axis motor set is connected located at X-axis synchrome conveying belt 113 and with X-axis guide pad 115.The Y-axis fortune Dynamic device includes：Y-axis guide rod 119, Y-axis synchrome conveying belt (not identifying in the drawings), Y-axis guide pad 121 and Y-axis electricity Machine (does not identify) in the drawings；Y-axis guide rod 119 is two, and relative the two of the top of frame 101 is respectively in along Y direction Side；Y-axis synchrome conveying belt is set parallel with Y-axis guide rod 119；Y-axis guide pad 121 is two, is sheathed on two Y-axis respectively Guide rod 119, and, the opposite end of X-axis guide rod 111 is inserted in two Y-axis guide pads 121 respectively；Y-axis motor is to be sheathed on Y-axis synchrome conveying belt and it is connected with Y-axis guide pad 121.In the above-described embodiments, the Y-axis in the Y-axis telecontrol equipment synchronously passes Send band and y-axis motor supporting, i.e. Y-axis synchrome conveying belt and y-axis motor can be one or two：An if (Y Axle synchrome conveying belt and a y-axis motor), this Y-axis synchrome conveying belt located at frame 101 top side and with that That Y-axis guide rod 119 in side is set parallel, and this supporting y-axis motor is arranged with this Y-axis synchrome conveying belt It is connected in this Y-axis synchrome conveying belt and with that Y-axis guide pad 121 in that side；If two (two Y-axis are same Walk conveyer belt x and two y-axis motors), the two Y-axis synchrome conveying belts are respectively in the opposite sides at the top of frame 101, and divide Do not set parallel with two Y-axis guide rods 119, two y-axis motor sets are respectively arranged on the two Y-axis synchrome conveying belts and right respectively That Y-axis guide pad 121 answered connects.The Z axis telecontrol equipment includes：Z axis guide rod 127, Z axis screw mandrel 129 and Z Spindle motor (is not identified) in the drawings, and Z axis guide rod 127 and Z axis screw mandrel 129 are set parallel along Z axis, and objective table 103 is arranged In Z axis guide rod 127 and Z axis screw mandrel 129, Z axis motor set is connected located at Z axis screw mandrel 129 and with objective table 103.In addition, X-axis Motor, y-axis motor and Z axis motor are both preferably stepper motor.
For above-mentioned preferred embodiment, X-axis telecontrol equipment, Y-axis telecontrol equipment in the three-dimensional motion mechanism and Z axis telecontrol equipment can still make other changes.For example, in a second embodiment, the X-axis telecontrol equipment includes：X-axis guide rod, X Axial filament bar, X-axis guide pad and X-axis motor, the X-axis guide rod and the X-axis screw mandrel are set parallel along X-axis, the number Code microscope is fixed on the X-axis guide pad and the X-axis guide pad is sheathed on the X-axis guide rod, and the X-axis motor is arranged It is connected in the X-axis screw mandrel and with the X-axis guide pad；The Y-axis telecontrol equipment includes：Y-axis guide rod, Y-axis screw mandrel, Y-axis Guide pad and y-axis motor；Y-axis guide rod is two, and the opposite sides at the top of the frame is respectively in along Y direction； The Y-axis screw mandrel is one or two, and is set parallel with the corresponding Y-axis guide rod；Y-axis guide pad is two, respectively Two Y-axis guide rods are sheathed on, and, the opposite end of the X-axis guide rod and the X-axis screw mandrel is inserted in two respectively The Y-axis guide pad；The y-axis motor for it is corresponding with the Y-axis screw mandrel one or two, be sheathed on corresponding to the Y-axis Screw mandrel and it is connected with the Y-axis guide pad；The Z axis telecontrol equipment includes：Z axis guide rod, Z axis screw mandrel and Z axis motor, The Z axis guide rod and the Z axis screw mandrel are set parallel along Z axis, and the objective table is sheathed on the Z axis guide rod and described Z axis screw mandrel, the Z axis motor set are connected located at the Z axis screw mandrel and with the objective table.In addition, X-axis motor, y-axis motor, And Z axis motor is both preferably stepper motor.
Control panel, it is connected with the three-dimensional motion mechanism and control terminal 20, for controlling X in the three-dimensional motion mechanism Spindle motor, y-axis motor and Z axis motor.In the present invention, the circuit board can be used based on STM32F103 chips YXRL_001 circuit control panels, on the one hand, the circuit control panel can pass through X-axis electricity in Du Pont's line and the three-dimensional motion mechanism Machine, y-axis motor and Z axis motor are electrically connected with, and on the other hand, the conversion that the circuit control panel can turn USB by a serial ports connects Head and be connected with control terminal 20, in this way, the circuit control panel can be just received from the control command that transmits of control terminal 20, To control X-axis motor in the three-dimensional motion mechanism, y-axis motor and Z axis motor.Specifically, in order that the electricity Road control panel can receive and perform the control command transmitted from control terminal 20, when necessary, it is also necessary to the circuit The firmware of control panel is changed accordingly, and needs to control the communication of terminal 20 to support.
It is described in detail below for the firmware operation of the YXRL_001 circuit control panels based on STM32F103 chips：
Firmware used in the circuit control panel is provided from STMicroelectronics (ST Microelectronics) STM32F10x chip standard firmware templates.In order to allow chip to respond the move sent from computer serial line interface, need Firmware template is modified, we define control instruction first, and the control instruction length of definition is unified for 5, and encodes Form is ASCII character (see the table below one).
The control instruction of table one
Control instruction Instruct effect
uXXXX The motor of Z axis is controlled to rotate forward XXXX units, objective table moves up
dXXXX The motor reversal XXXX units of Z axis are controlled, objective table moves down
rXXXX The motor of X-axis is controlled to rotate forward XXXX units, shower nozzle platform moves right
lXXXX The motor reversal XXXX units of X-axis are controlled, shower nozzle platform is moved to the left
fXXXX The motor of Y-axis is controlled to rotate forward XXXX units, shower nozzle platform moves forward
bXXXX The motor reversal XXXX units of Y-axis are controlled, shower nozzle platform is moved rearwards
Wherein, XXXX 0000--9999, represent that stepper motor rotates how many individual units
, it is necessary to change firmware template after control instruction is defined so that it can be received sends from control terminal 20 Instruction.It is to take the mode sent by turn during due to serial communication interface transmission data, we adapt firmware so that it is often received Single treatment is carried out to 5 message.Motor (X-axis motor, y-axis motor and Z axis are calculated by simple string processing Motor) after the units that rotates, make motor carry out the rotation of corresponding unit to respective direction.The reorganization of firmware completes it Afterwards, by corresponding software, the firmware after reorganization is written in the circuit control panel using cd-rom recorder.
For above-mentioned objective table 103, more use rectangular mesa or the triangle table top of objective table 103, the one of objective table 103 Individual side is as the connection side being connected with the Z axis telecontrol equipment.In a preferred embodiment：In the Z axis telecontrol equipment In, including two Z axis guide rods 127 and a Z axis screw mandrel 129, wherein, Z axis screw mandrel 129 is placed in the middle, two Z axis guide rods 127 Positioned at the opposite sides of Z axis screw mandrel 129；Objective table 103 is preferably triangle table top, then one side of objective table 103 is as connecting side Side, the both ends of the connection side are sheathed on two Z axis guide rods 127, and the middle part of the connection side is then socketed on that Z Axial filament bar 129.In addition, objective table 103 is placed on a luggage carrier 102, the shape of luggage carrier 102 is and objective table 103 Shape it is consistent, as it was previously stated, objective table 103 be preferably triangle table top, luggage carrier 102 be preferably also tripod, as load Three corners of the tripod of rack 102 are equipped with adjustable knob, and the adjustable knob is made up of tightening screw and spring, screwed Screw is attached to corner portion connection protrusive board, and spring housing is located at the tightening screw and contacts the objective table 103 in top, screws spiral shell The bottom of nail is additionally provided with knob, in this way, can carry out leveling to objective table 103 by screwing and unclamping the knob.
Data microscope 105 is turned again to, in the present embodiment, digit microscope 105 is fixed on X-axis guide pad 115 On, two-dimensional movement can be carried out in X-axis and Y-axis with X-axis guide pad 115.In addition, digit microscope 105 can pass through wired biography Transferring technology (such as USB transmission line) with control terminal 20 (such as PC PC, laptop Notebook or work Stand) establish communication connection.Digit microscope 105 is located at the top of objective table 103, is seen underface by built-in shooting function The micro- effect of nematode in the culture of nematodes ware 30 measured preserves and forms image information, by analog-to-digital conversion, by mould Number converted images information transmission is to controlling terminal 20 and being shown on the display screen of control terminal, by controlling terminal The micro image analysis software installed on 20 is tracked analysis, carries out image recognition processing to complete the identification of nematode and determine A series of position, so as to obtain valuable qualitative, quantitative data.Further, to cause digit microscope 105 to be fixed on X-axis guide pad More firm on 115, in a preferred embodiment, digit microscope 105 is by a microscope mounting bracket 107 (as shown in Figure 3) And it is fixed on X-axis guide pad 115.From the figure 3, it may be seen that microscope mounting bracket 107 includes base 104, offered on base 104 Observation panel 106, for assembling the camera lens of digit microscope 105, to cause the camera lens can pass through observation panel 104 to observe under it Nematode on objective table 103 in nematode culture dish 30, the spider 108 set up through support frame is also included on base 104, for binding round Firmly digit microscope 105, play a part of spacing and firm digit microscope 105.
It should be noted that in the present embodiment, digit microscope 105 is fixed on X-axis guide pad 115 and permeable described three Tie up motion and adjust with the relative position of objective table 103 to be carried out to the nematode in nematode culture dish 30 on objective table 103 Observation and positioning.To lift observing effect, in the present invention, observation light source also is provided specifically for culture of nematodes ware 30, it is described Observation light source is located at the bottom of the culture of nematodes ware to provide the uniform parallel light source towards culture of nematodes ware 30.It is optional one In embodiment, the observation light source (not shown in the drawings) is LED multi-point sources, and the LED multi-point sources are placed in On the objective table, a frosted glass is put on the LED multi-point sources, culture of nematodes ware 30 is placed in the frosted glass On, in this way, with LED multi-point sources frosted glass combination so that digit microscope 105 more can be observed clearly The nematode into culture of nematodes ware 30.Preferably, to cause the LED multi-point sources to produce source of parallel light as far as possible, it is desirable to which LED is more Each LED itself size in spot light wants the spacing between small and adjacent LED lamp as small as possible, in the certain feelings of area The LED gathered under shape will be as more as possible, in this way, when the visual field of digit microscope 105 corresponds to culture of nematodes ware 30, it is described Some light emitting diode during LED multi-point sources are put can be right against digit microscope 105, and light can be vertically from nematode It is irradiated to come immediately below culture dish 30, may be considered substantially in the visual field uniformly, nematode can be seen.Further, I A convex lens can be being put above light source, allow all LED all on the focal plane of convex lens, then by culture dish frame convex Directly over lens (if but use plus the method for convex lens is, it is necessary to the less magnifying glass of focal length because wanting one and original big The same uniformly light-emitting region of small area, it would be desirable to which culture dish and back light source intermediate distance are 2 times of focal lengths).
Control terminal 20, digit microscope 105 and control panel (not shown in the drawings) communication connection.It is preferred one In embodiment, control terminal 20 is, for example, PC PC, laptop Notebook or work station etc..Control terminal 20 receive at digit microscope 105 transmit come on the image information observed by digit microscope 105 and by figure As being shown on the display screen after carrying out corresponding identifying processing, according to the image information of display, terminal 20 is controlled to the control Making sheet exports the control instruction for controlling the three-dimensional motion mechanism, digital corresponding to the three-dimensional motion mechanism adjustment to make Relative position between microscope 105 and objective table 103, realizes auto-focusing so that digit microscope 105 is it is observed that nematode Nematode in culture dish 30 simultaneously obtains the nematode image information that can meet image recognition requirement, and by control terminal 20 according to described Nematode image information carries out image recognition to identify the nematode in the nematode image information.
It is described in detail below for auto-focusing part：
In the present embodiment, we evaluate letter from squared gradient function (formula (1) in seeing below) as definition Count to calculate the definition of image：
When the maximum of sharpness evaluation function is found by moving up and down objective table 103, it is meant that Alignment is burnt.
In the focusing application of reality, gray level image need to be processed the image into.It is transmitted across in order to reduce picture signal Influence, is smoothed to image, is gaussian filtering (window herein, used by smoothing processing caused by noise in journey 33) mouth size is, for example,.
When culture of nematodes ware 30 is with the distance and the larger range deviation for being directed at Jiao of digit microscope 105, scheme in the visual field The gradient of picture is smaller, the variation unobvious of sharpness evaluation function when moving up and down objective table 103, now, definition evaluation Function does not just play effect.In view of this, in the present embodiment, the three-dimensional motion is controlled first with the circuit control panel X-axis motor and y-axis motor planar Z axis in mobile digital microscope 105 and the control three-dimensional motion mechanism in mechanism Motor moves up and down objective table 103 so that digit microscope 105 can find the place of nematode and substantially see (fuzzy ) image, most clearly image is then obtained by the algorithm of auto-focusing again.
In theory, sharpness evaluation function is unimodal, i.e. head with the function of distance change between camera lens and shooting object First can with the increase of distance monotonic increase, reach after peak value monotone decreasing again.In view of this, it is necessary first to detect to work as Front position is to be in monotonic increase state or monotone decreasing state, then controls digit microscope 105 and culture of nematodes ware 30 The distance between each time with fixed step-length to peak value direction move.(after being incremented by and rolled over when the state of dullness is broken It is curved to successively decrease), illustrate to have had passed over peak value, at this point it is possible to utilize the thought of similar dichotomy, relatively current location is with being somebody's turn to do every time The size of the value of the sharpness evaluation function of above current location and current location lower section equidistant position is (i.e.：Current location Sharpness evaluation function value Sc, the sharpness evaluation function value S of the position of one determining deviation of current location topc-Δd, present bit Put the sharpness evaluation function value S of the position of one determining deviation of lower sectionc+Δd), then move in three at maximum, then shorten one Half displacement.It steps be repeated alternatively until that displacement reaches the minimum value that can be moved, now, it is assumed that to have focused Into.During above-mentioned mobile control, in order to calculate the value of the sharpness evaluation function after movement, it is necessary to wait mobile end And image stabilization, so a time interval (such as 1500 milliseconds) preferably can be set in mobile and last movement every time. In this time interval, still to continue to image and show the visual field, the control of time interval can be by a timer come real It is existing.
Further referring to Fig. 4, it is the functional block diagram that terminal 20 is controlled in nematode identifying system of the present invention.Such as figure Shown in 4, control terminal 20 further comprises：Gray proces unit 201, smoothing processing unit 203, self-adaption thresholding processing are single Member 205 and contours extract unit 207.Gray proces unit 201, for carrying out gray proces to the nematode image information. Smoothing processing unit 203 is connected with gray proces unit 201, for the institute after the progress gray proces of gray proces unit 201 Nematode image information is stated to be smoothed.Self-adaption thresholding processing unit 205 is connected with smoothing processing unit 203, is used for The nematode image information after being smoothed to smoothing processing unit 203 carries out self-adaption thresholding processing.Profile carries Unit 207 is taken to be connected with self-adaption thresholding processing unit 205, it is adaptive for being carried out to self-adaption thresholding processing unit 205 The nematode image information after thresholding processing is answered to carry out contours extract.
In a preferred embodiment：
Gray proces are carried out to the nematode image information, nematode image information progress gray proces can be caused Image is more prone to processed and calculated, by taking OpenCV (Open Source Computer Vision Library) as an example, because To be stored in colour picture among matrix in the form of 83 passages in OpenCV, i.e., each pixel saves as 0 and arrived 3 integers between 255, R are corresponded to respectively, tri- kinds of colors of G, B, after gray proces are carried out, each pixel can save as 0 and arrive An integer between 255, image are then changed into 8 single pass images.
The nematode image information after gray proces is smoothed, the target of nematode identification is to identify the visual field In big nematode, the eelworm in image typically in 1mm to 2mm, in identification, is regarded as noise by the length of big nematode, first right Image is smoothed (also referred to as Fuzzy Processing), and more commonly used smoothing processing method is to use medium filtering.
Image recognition processing is carried out to the nematode image information to complete the identification of nematode, at self-adaption thresholding The identification method that reason and contours extract are combined, is specifically included：A kind of algorithm of automatic calculating threshold value is provided, and needs this Threshold value can be different with the region value of brightness in different colours.Briefly, algorithm used herein is：Using each pixel as Center, the pixel in around the pixel one square area delimited is averaged to determine the adaptive threshold at this point (for example, can be averaged to the pixel in around the pixel 21 × 21 square inner region and then the average value is subtracted into 7 Determine the adaptive threshold at this point)；During processing, the pixel placement that will be above the adaptive threshold is 0, and will be less than this certainly The pixel placement for adapting to threshold value is 255.After the result of adaptive threshold is obtained (as shown in Figure 5), by extracting in image most The outline of outside finds the pixel on the border of all white portions, calculates the area that each outline is included and finds out that A little areas exceed the profile of predefined size, delete be not nematode the white portion no more than predefined size, area is exceeded in advance Determining the white portion of size, (for example, in the case of screen resolution is 640 × 480, we can will be pre- as target area Determine size and be set as 1000, in this way, in actual applications, exactly calculating the area that each outline is included and finding out those faces Profile of the product more than 1000, deleted be not nematode those be no more than 1000 small white portion, area is more than 1000 White portion as target area), sketch out rectangular bounding box and the circumscribed circle of these target areas to mark in the picture Their position (as shown in Figure 6).
Nematode identifying system provided by the invention, including organic frame, objective table, digit microscope and control terminal, institute Control terminal is stated to transmit the nematode image information come at the digit microscope and carry out image to the nematode image information Identification, to identify the nematode in the nematode image information.Compared to prior art, nematode identifying system of the present invention, which has, to be known Not simple in construction, identification is quick and nematode discrimination is high and versatility is good easy to utilize, can have miscellaneous bacteria and illumination not The advantages that being identified in uniform environment.
Present invention also offers a kind of nematode recognition methods based on above-mentioned nematode identifying system, referring to Fig. 7, showing The schematic flow sheet of nematode recognition methods of the present invention in one embodiment.With reference to Fig. 1 to Fig. 7, nematode recognition methods of the present invention Comprise the following steps：
Step S51, nematode identifying system is debugged in advance.In a preferred embodiment, debugging nematode identifying system may include But it is not limited to herein below：The connection of digit microscope 105, digit microscope 105 and control panel is installed, to the control Plate carries out firmware operation, debugs three-dimensional motion mechanism.
In the present invention, the three-dimensional motion mechanism is mainly used for adjusting digit microscope 105 and objective table 103 is relative Position relationship, so that digit microscope 105 can more clearly obtain the figure of nematode in nematode culture dish 30 on objective table 103 As information in favor of observing and identify.In a preferred embodiment, the three-dimensional motion mechanism includes：For driving number The X-axis telecontrol equipment that microscope 105 moves along X-axis；For the Y-axis telecontrol equipment for driving digit microscope 105 to be moved along Y-axis； And the Z axis telecontrol equipment moved for driving objective table 103 along Z axis.For above-mentioned preferred embodiment, the X-axis motion Device includes：X-axis guide rod 111, X-axis synchrome conveying belt 113, X-axis guide pad 115 and X-axis motor, X-axis guide rod 111 Set parallel along X-axis with X-axis synchrome conveying belt 113, X-axis guide pad 115 is sheathed on the X-axis guide rod, digit microscope 105 are fixed on X-axis guide pad 115, and X-axis motor set is connected located at X-axis synchrome conveying belt 113 and with X-axis guide pad 115.Institute Stating Y-axis telecontrol equipment includes：Y-axis guide rod 119, Y-axis synchrome conveying belt, Y-axis guide pad 121 and y-axis motor；Y-axis is oriented to Bar 119 is two, and the opposite sides at the top of frame 101 is respectively in along Y direction；Y-axis synchrome conveying belt is oriented to Y-axis Bar 119 is set parallel；Y-axis guide pad 121 is two, is sheathed on two Y-axis guide rods 119 respectively, and, X-axis guide rod 111 Opposite end is inserted in two Y-axis guide pads 121 respectively；Y-axis motor be sheathed on Y-axis synchrome conveying belt and with Y-axis guide pad 121 connections.In the above-described embodiments, the Y-axis synchrome conveying belt in the Y-axis telecontrol equipment and y-axis motor are supporting, i.e. Y-axis Synchrome conveying belt and y-axis motor can be one or two：If (a Y-axis synchrome conveying belt x and a Y-axis electricity Machine), this Y-axis synchrome conveying belt located at frame 101 top side and with that Y-axis guide rod in that side 119 parallel settings, this supporting y-axis motor is sheathed on this Y-axis synchrome conveying belt with this Y-axis synchrome conveying belt And it is connected with that Y-axis guide pad 121 in that side；If two (two Y-axis synchrome conveying belt x and two Y-axis electricity Machine), the two Y-axis synchrome conveying belts are respectively in the opposite sides at the top of frame 101, and respectively with two Y-axis guide rods 119 Parallel to set, two y-axis motor sets are respectively arranged on the two Y-axis synchrome conveying belts and respectively that corresponding Y-axis guide pad 121 connections.The Z axis telecontrol equipment includes：Z axis guide rod 127, Z axis screw mandrel 129 and Z axis motor, Z axis guide rod 127 Set parallel along Z axis with Z axis screw mandrel 129, objective table 103 is sheathed on Z axis guide rod 127 and Z axis screw mandrel 129, Z axis motor set It is connected located at Z axis screw mandrel 129 and with objective table 103.In addition, X-axis motor, y-axis motor and Z axis motor are both preferably stepping Motor.
Digit microscope 105 is installed, refers to digit microscope 105 being installed on the top of objective table 103.It is preferred real one Apply in example, it is fixed on digit microscope 105 on the X-axis guide pad 115 of the top of objective table 103, so, digit microscope 105 can carry out two-dimensional movement with X-axis guide pad 115 in X-axis and Y-axis.To cause digit microscope 105 to be fixed on X-axis More firm on guide pad 115, in a preferred embodiment, digit microscope 105 is fixed by a microscope mounting bracket 107 In on X-axis guide pad 115.Microscope mounting bracket 107 includes base 104, observation panel 106 is offered on base 104, for filling Camera lens with digit microscope 105, to cause the permeable observation panel 106 of the camera lens to observe nematode on the objective table 103 under it Nematode in culture dish 30, the spider 108 set up through support frame is also included on base 104, for waling digit microscope 105, play a part of spacing and firm digit microscope 105.
The connection of digit microscope 105 and control panel, refer to the connection for digit microscope 105 and control The connection of making sheet.The connection of digit microscope 105 is specifically included by data wire (such as USB transmission line) and control Terminal 20 (such as PC PC, laptop Notebook or work station etc.) establishes communication connection, certainly, circuit Connection be also not limited to data wire, can also the transmission technology such as wireless blue tooth or wireless WiFi with control terminal 20 foundation communicate Connection.The connection of control panel specifically include by X-axis motor in Du Pont's line and the three-dimensional motion mechanism, y-axis motor, with And Z axis motor is electrically connected with and turns USB crossover sub with controlling terminal 20 to be connected by a serial ports.
Firmware operation is carried out to the control panel：In order that the control panel can be received and performed from control terminal 20 The control command transmitted, when necessary, it is also necessary to changed accordingly the firmware of the circuit control panel, and need to control The communication of terminal 20 processed is supported.In a preferred embodiment, the control panel can use the YXRL_ based on STM32F103 chips 001 circuit control panel, illustrate for the firmware operation of the YXRL_001 circuit control panels based on STM32F103 chips before can referring to The associated description stated, will not be repeated here.
Debug three-dimensional motion mechanism：Graphical operation and control interface for three-dimensional motion mechanism is provided in control terminal 20, The graphical operation and control interface show " left side " and " right side " of X-axis in corresponding three-dimensional motion, correspondence Y-axis " preceding " and The parameters such as "up" and "down" of " rear " and corresponding Z axis, by user on graphical operation and control interface in corresponding parameter field it is defeated Enter required displacement numerical value, control terminal 20 can export phase according to these parameters and its corresponding displacement numerical value The control instruction answered simultaneously is sent to the control panel, to make the control panel control the X-axis in the three-dimensional motion mechanism accordingly One or more of motor, y-axis motor and Z axis motor are accordingly operated and realize the shifting of corresponding displacement numerical value It is dynamic.In addition, as it was previously stated, digit microscope 105 is affixed on the X-axis guide pad, therefore, can also be by described in control Three-dimensional motion mechanism realizes the automatic focusing function of digit microscope, specifically：Opened on the display screen of control terminal 20 Qi Xin display window shows the image of digit microscope 105, by controlling the Z axis motor in the three-dimensional motion mechanism to adjust The relative position of the whole digit microscope and the objective table, so as to realize the function of auto-focusing.
Step S53, the culture of nematodes ware 30 that culture has nematode is positioned at observation position.In the present embodiment, step S53 includes：The culture of nematodes ware 30 that has nematode will be cultivated to be positioned on objective table 103 and provide observation for culture of nematodes ware 30 Light source.In actual applications, culture of nematodes ware 30 can be positioned on objective table 103 by certain mode or instrument.Such as preceding institute State, objective table 103 is preferably triangle table top, and a side of objective table 103 is as the connection being connected with the Z axis telecontrol equipment Side.In a preferred embodiment：In the Z axis telecontrol equipment, including two Z axis guide rods 127 and led positioned at two Z axis To a Z axis screw mandrel 129 between bar 127, the both ends of the connection side are sheathed on two Z axis guide rods 127, the connection The middle part of side is then socketed on that Z axis screw mandrel 129.In addition, objective table 103 is placed on a luggage carrier 102, loading The shape of frame 102 is, preferably tripod consistent with the shape of objective table 103, three as the tripod of luggage carrier 102 Corner is equipped with adjustable knob, and the adjustable knob is made up of tightening screw and spring, and tightening screw is attached to corner portion connection Protrusive board, spring housing is located at the tightening screw and contacts the objective table 103 in top, and the bottom of tightening screw is additionally provided with knob, In this way, leveling can be carried out to objective table 103 by screwing and unclamping the knob.
To lift observing effect, in the present invention, also observation light source, the observation are provided specifically for culture of nematodes ware 30 Light source is located at the bottom of the culture of nematodes ware to provide the uniform parallel light source towards culture of nematodes ware 30.In an optional implementation In example, the observation light source (not shown in the drawings) is LED multi-point sources, and the LED multi-point sources are placed in described On objective table, a frosted glass is put on the LED multi-point sources, culture of nematodes ware 30 is placed on the frosted glass, such as This, the combination of frosted glass with LED multi-point sources, so that digit microscope 105 more can clearly observe line Nematode in worm culture dish 30.Further, we can put a convex lens above light source, allow all LED all in convex lens On the focal plane of mirror, then by culture dish frame directly over convex lens (if but use plus convex lens method, it is necessary to which focal length is smaller Magnifying glass because want a uniformly light-emitting region as original area, it would be desirable to culture dish and back light source Intermediate distance is 2 times of focal lengths).
Therefore,, can also root after the culture of nematodes ware 30 that culture has nematode is placed in into objective table 103 in step S53 Require according to observation and objective table 103 is adjusted using the adjustable knob in three corners of the tripod as luggage carrier 102 Put down operation and the exit positions of LED multi-point sources are adjusted according to the radiation response of light source and go out luminous intensity.
Step S55, the nematode on objective table 103 in nematode culture dish 30 is observed using digit microscope 105, and with reference to three Motion is tieed up to adjust the relative position of digit microscope 105 and objective table 103, obtains the line that can meet image recognition requirement Worm image information.In actual applications：Digit microscope 105 is adjusted on objective table 103 using the three-dimensional motion mechanism The surface of culture of nematodes ware 30, so that the nematode in digit microscope 105 and culture of nematodes ware 30 realizes focusing and can be to line Nematode in worm culture dish 30 is observed；Being trained on nematode on objective table 103 observed by being obtained by digit microscope 105 The image information of nematode in ware 30 is supported, by described image information transmission to the control terminal, by the display of the control terminal Screen shows described image information in display window；By it is described control terminal according to described image information and output control refers to Make (such as:On graphical operation and control interface required displacement numerical value is inputted in corresponding parameter field) and by the control Making sheet come control one or more of X-axis motor, y-axis motor and Z axis motor in the three-dimensional motion mechanism carry out phase Should operate to adjust the relative position of digit microscope 105 and objective table 103, pass through such observed image -- three-dimensional motion machine The circulation step of structure adjustment -- observed image -- three-dimensional motion mechanism adjustment, it is final to obtain energy by digit microscope 105 Meet the nematode image information of image recognition requirement.In this step, it is preferable that the three-dimensional motion mechanism includes：Driving number The X-axis telecontrol equipment that code microscope 105 moves along X-axis, the Y-axis telecontrol equipment that driving digit microscope 105 moves along Y-axis, with And the Z axis telecontrol equipment that driving objective table moves along Z axis, therefore, control the three-dimensional motion mechanism to adjust digit microscope 105 and the relative position of objective table 103 refer to：Adjust fortune of the digit microscope 105 in the plane that X-axis and Y-axis are formed Motion dynamic and that adjustment objective table 103 is on Z axis.Certainly it is not limited thereto, in other embodiments, if three maintenance and operation Motivation structure includes：For the X-axis telecontrol equipment for driving digit microscope 105 to be moved along X-axis；For driving digit microscope 105 The Y-axis telecontrol equipment moved along Y-axis；And the Z axis telecontrol equipment for driving digit microscope 105 to be moved along Z axis, then, and control The three-dimensional motion mechanism is made to adjust the relative position of the digit microscope and the objective table, including：Adjust the number Three-dimensional motion of the code microscope in X-axis, Y-axis and Z axis.
Realize that the part of focusing is retouched in detail with the nematode in culture of nematodes ware 30 below for digit microscope 105 State：
When the maximum of sharpness evaluation function is found by moving up and down objective table 103, it is meant that Alignment is burnt.
In the focusing application of reality, gray level image need to be processed the image into.It is transmitted across in order to reduce picture signal Influence, is smoothed to image, is gaussian filtering (window herein, used by smoothing processing caused by noise in journey 33) mouth size is, for example,.
When culture of nematodes ware 30 is with the distance and the larger range deviation for being directed at Jiao of digit microscope 105, scheme in the visual field The gradient of picture is smaller, the variation unobvious of sharpness evaluation function when moving up and down objective table 103, now, definition evaluation Function does not just play effect.In view of this, in the present embodiment, the three-dimensional motion is controlled first with the circuit control panel X-axis motor and y-axis motor planar Z axis in mobile digital microscope 105 and the control three-dimensional motion mechanism in mechanism Motor moves up and down objective table 103 so that digit microscope 105 can find the place of nematode and substantially see (fuzzy ) image, most clearly image is then obtained by the algorithm of auto-focusing again.
In theory, sharpness evaluation function is unimodal, i.e. head with the function of distance change between camera lens and shooting object First can with the increase of distance monotonic increase, reach after peak value monotone decreasing again.In view of this, it is necessary first to detect to work as Front position is to be in monotonic increase state or monotone decreasing state, then controls digit microscope 105 and culture of nematodes ware 30 The distance between each time with fixed step-length to peak value direction move.(after being incremented by and rolled over when the state of dullness is broken It is curved to successively decrease), illustrate to have had passed over peak value, at this point it is possible to utilize the thought of similar dichotomy, relatively current location is with being somebody's turn to do every time The size of the value of the sharpness evaluation function of above current location and current location lower section equidistant position is (i.e.：Current location Sharpness evaluation function value Sc, the sharpness evaluation function value S of the position of one determining deviation of current location topc-Δd, present bit Put the sharpness evaluation function value S of the position of one determining deviation of lower sectionc+Δd), then move in three at maximum, then shorten one Half displacement.It steps be repeated alternatively until that displacement reaches the minimum value that can be moved, now, it is assumed that to have focused Into.During above-mentioned mobile control, in order to calculate the value of the sharpness evaluation function after movement, it is necessary to wait mobile end And image stabilization, so a time interval (such as 1500 milliseconds) preferably can be set in mobile and last movement every time. In this time interval, still to continue to image and show the visual field, the control of time interval can be by a timer come real It is existing.
Step S57, image recognition is carried out to the nematode image information, to identify the line in the nematode image information Worm.
Referring to Fig. 8, show the schematic flow sheet of the further refinement steps of step S57.As shown in figure 8, step S57 Further comprise：
Step S571, gray proces are carried out to the nematode image information.The nematode image information is carried out at gray scale Reason, image can be caused to be more prone to processed and calculate nematode image information progress gray proces, with OpenCV Exemplified by (Open Source Computer Vision Library), because by colour picture with 83 passages in OpenCV Form be stored among matrix, i.e., each pixel saves as 3 integers between 0 to 255, corresponds to R, G, B tri- respectively Kind color, after gray proces are carried out, each pixel can save as an integer between 0 to 255, and image is then changed into 8 Single pass image.
Step S573, the nematode image information after gray proces is smoothed.To the institute after gray proces State nematode image information to be smoothed, the target of nematode identification is to identify the big nematode in the visual field, the length of big nematode Typically in 1mm to 2mm, in identification, the eelworm in image is regarded as noise, first image is smoothed (also referred to as Fuzzy Processing), more commonly used smoothing processing method is to use medium filtering.
Step S575, self-adaption thresholding processing is carried out to the nematode image information after smoothing processing.It is preferred one In implementation, there is provided a kind of algorithm of automatic calculating threshold value, and need this threshold value to be taken in the region of different colours and brightness Value is different.Specifically, the algorithm for calculating threshold value automatically is：Centered on each pixel, delimit around the pixel one Pixel in square area average determine at this point adaptive threshold (for example, can to around the pixel 21 × Pixel in 21 square inner region is averaged and then the average value is subtracted into 7 to determine the adaptive thresholding at this point Value)；During processing, the pixel placement that will be above the adaptive threshold is 0, and the pixel placement that will be less than the adaptive threshold is 255, so as to obtain the result of adaptive threshold (as shown in Figure 5).
Step S577, contours extract is carried out to the nematode image information after self-adaption thresholding processing.Continue from the preceding paragraph, After the result of adaptive threshold is obtained, the border of all white portions is found by extracting in image outmost outline Pixel, calculate the area that each outline is included and find out those areas exceed predefined size profile, it is not line to delete The white portion no more than predefined size of worm, using area exceed predefined size white portion as target area (for example, In the case of screen resolution is 640 × 480, predefined size can be set as 1000 by us, in this way, in actual applications, Exactly calculate the area that each outline is included and find out profile of those areas more than 1000, deleted be not nematode that A little small white portions no more than 1000, using white portion of the area more than 1000 as target area), sketch out these mesh The rectangular bounding box and circumscribed circle for marking region mark their position (as shown in Figure 6) in the picture.
By above-mentioned steps S571 to step S577, the nematode in the nematode image information can be efficiently identified out.
Nematode recognition methods provided by the invention, by the nematode in culture of nematodes ware described in digital micrograph sem observation and is obtained The nematode image information of image recognition requirement can be met by taking, to the nematode image information carry out gray proces, smoothing processing, from After adapting to thresholding processing and contours extract, the nematode in the nematode image information is identified.Compared to prior art, Nematode recognition methods of the present invention has identification method simple and quickly, nematode discrimination is high and versatility is good easy to utilize, It can be identified in having the advantages that miscellaneous bacteria and the even environment of uneven illumination.So the present invention effectively overcomes prior art In various shortcoming and have high industrial utilization.
The above-described embodiments merely illustrate the principles and effects of the present invention, not for the limitation present invention.It is any ripe Know the personage of this technology all can carry out modifications and changes under the spirit and scope without prejudice to the present invention to above-described embodiment.Cause This, those of ordinary skill in the art is complete without departing from disclosed spirit and institute under technological thought such as Into all equivalent modifications or change, should by the present invention claim be covered.

Claims (19)

  1. A kind of 1. nematode identifying system, it is characterised in that including：
    Frame；
    Objective table in the frame, for configuring nematode culture dish；
    Digit microscope in the frame and above the objective table, for observing in the culture of nematodes ware Nematode simultaneously obtains the nematode image information that can meet image recognition requirement；And
    Terminal is controlled, is communicated to connect with the digit microscope, the nematode of coming is transmitted at the digit microscope for receiving Image information simultaneously carries out image recognition to the nematode image information；The control terminal includes：Gray proces unit, for pair The nematode image information carries out gray proces；Smoothing processing unit, it is connected with the gray proces unit, for the ash The nematode image information after degree processing unit progress gray proces is smoothed；Self-adaption thresholding processing unit, It is connected with the smoothing processing unit, for the nematode image information after being smoothed to the smoothing processing unit Carry out self-adaption thresholding processing；Contours extract unit, be connected with the self-adaption thresholding processing unit, for it is described from Adapt to thresholding processing unit and carry out the nematode image information progress contours extract after self-adaption thresholding processing, with identification The nematode gone out in the nematode image information.
  2. 2. nematode identifying system as claimed in claim 1, it is characterised in that also include the observation of the corresponding culture of nematodes ware Light source, the observation light source are to provide the uniform parallel towards the culture of nematodes ware positioned at the bottom of the culture of nematodes ware Light source.
  3. 3. nematode identifying system as claimed in claim 2, it is characterised in that the uniform parallel light source includes LED multiple spot light Source, the LED multi-point sources are placed on the objective table, and a frosted glass, the nematode are configured with the LED multi-point sources Culture dish is placed on the frosted glass.
  4. 4. nematode identifying system as claimed in claim 1, it is characterised in that also include：
    Three-dimensional motion machine in the frame, for adjusting the digit microscope and the objective table relative position relation Structure；And
    The control panel being connected with the three-dimensional motion mechanism and the control terminal, for receiving the mobile end from communication connection The control instruction at end and control the three-dimensional motion mechanism.
  5. 5. nematode identifying system as claimed in claim 4, it is characterised in that the three-dimensional motion mechanism includes：
    The X-axis telecontrol equipment for driving the digit microscope to be moved along X-axis；
    The Y-axis telecontrol equipment for driving the digit microscope to be moved along Y-axis；And
    The Z axis telecontrol equipment for driving the objective table to be moved along Z axis.
  6. 6. nematode identifying system as claimed in claim 5, it is characterised in that
    The X-axis telecontrol equipment includes：X-axis guide rod, X-axis screw mandrel, X-axis guide pad and X-axis motor, the X-axis guide rod Set parallel along X-axis with the X-axis screw mandrel, the digit microscope is fixed on the X-axis guide pad and the X-axis guide pad The X-axis guide rod is sheathed on, the X-axis motor set is connected located at the X-axis screw mandrel and with the X-axis guide pad；
    The Y-axis telecontrol equipment includes：Y-axis guide rod, Y-axis screw mandrel, Y-axis guide pad and y-axis motor；Y-axis guide rod is two It is individual, the opposite sides at the top of the frame is respectively in along Y direction；The Y-axis screw mandrel and a Y-axis guide rod are simultaneously Row is set；Y-axis guide pad is two, is sheathed on two Y-axis guide rods respectively, and, the X-axis guide rod and the X-axis The opposite end of screw mandrel is inserted in two Y-axis guide pads respectively；The y-axis motor be sheathed on a Y-axis screw mandrel and It is connected with a Y-axis guide pad；And
    The Z axis telecontrol equipment includes：Z axis guide rod, Z axis screw mandrel and Z axis motor, the Z axis guide rod and the Z axis Screw mandrel is set parallel along Z axis, and the objective table is sheathed on the Z axis guide rod and the Z axis screw mandrel, the Z axis motor set It is connected located at the Z axis screw mandrel and with the objective table.
  7. 7. nematode identifying system as claimed in claim 5, it is characterised in that
    The X-axis telecontrol equipment includes：X-axis guide rod, X-axis synchrome conveying belt, X-axis guide pad and X-axis motor, the X-axis Guide rod and the X-axis synchrome conveying belt are set parallel along X-axis, the digit microscope be fixed on the X-axis guide pad and The X-axis guide pad is sheathed on the X-axis guide rod, the X-axis motor set located at the X-axis synchrome conveying belt and with the X Axle guide pad connects；
    The Y-axis telecontrol equipment includes：Y-axis guide rod, Y-axis synchrome conveying belt, Y-axis guide pad and y-axis motor；Y-axis is oriented to Bar is two, and the opposite sides at the top of the frame is respectively in along Y direction；The Y-axis synchrome conveying belt and an institute Y-axis guide rod is stated to set parallel；Y-axis guide pad is two, is sheathed on two Y-axis guide rods respectively, and, the X-axis is led Two Y-axis guide pads are inserted in respectively to the opposite end of bar；The y-axis motor is sheathed on a Y-axis and synchronously passed Send band and be connected with a Y-axis guide pad；And
    The Z axis telecontrol equipment includes：Z axis guide rod, Z axis screw mandrel and Z axis motor, the Z axis guide rod and the Z axis Screw mandrel is set parallel along Z axis, and the objective table is sheathed on the Z axis guide rod and the Z axis screw mandrel, the Z axis motor set It is connected located at the Z axis screw mandrel and with the objective table.
  8. 8. nematode identifying system as claimed in claims 6 or 7, it is characterised in that the digit microscope is micro- by one Mirror mounting bracket and be fixed on the X-axis guide pad.
  9. 9. nematode identifying system as claimed in claims 6 or 7, it is characterised in that
    The objective table is rectangular mesa or triangle table top, a side of the objective table as with the Z axis telecontrol equipment The connection side of connection；And
    In the Z axis telecontrol equipment, including two Z axis guide rods and a Z axis screw mandrel, the Z axis screw mandrel occupy In, two Z axis guide rods are located at the opposite sides of the Z axis screw mandrel.
  10. 10. nematode identifying system as claimed in claims 6 or 7, it is characterised in that the X-axis motor, the y-axis motor, with And the Z axis motor is stepper motor.
  11. 11. a kind of nematode recognition methods of the nematode identifying system based on as any one of claim 1 to 10, its feature It is, including：
    The culture of nematodes ware that culture has nematode is configured at objective table；
    Nematode in the culture of nematodes ware as described in digital micrograph sem observation simultaneously obtains the nematode image that can meet image recognition requirement Information；And
    The nematode image information is received by control terminal and image recognition is carried out to the nematode image information, including：To institute State nematode image information and carry out gray proces；The nematode image information after gray proces is smoothed；It is and right The nematode image information after smoothing processing carries out self-adaption thresholding processing and contours extract, to identify that the nematode is schemed As the nematode in information.
  12. 12. nematode recognition methods as claimed in claim 11, it is characterised in that configure the culture of nematodes ware that culture has nematode In objective table, including：
    The culture of nematodes ware that culture has nematode is positioned on objective table；And
    Observation light source is provided for the culture of nematodes ware.
  13. 13. nematode recognition methods as claimed in claim 12, it is characterised in that the observation light source is to be trained positioned at the nematode The bottom of ware is supported, to provide the uniform parallel light source towards the culture of nematodes ware.
  14. 14. nematode recognition methods as claimed in claim 12, it is characterised in that also include carrying out leveling to the objective table.
  15. 15. nematode recognition methods as claimed in claim 11, it is characterised in that the culture of nematodes as described in digital micrograph sem observation Nematode in ware simultaneously obtains the nematode image information that can meet image recognition requirement, including：
    The image information on nematode in the culture of nematodes ware on the objective table observed is obtained by digit microscope, By described image information transmission to the control terminal with digit microscope communication connection；
    By it is described control terminal according to described image information and output control instruct and three-dimensional motion machine is controlled by control panel Structure is to adjust the relative position of the digit microscope and the objective table, so as to realize auto-focusing；And
    The nematode image information of image recognition requirement can be met by being obtained by the digit microscope.
  16. 16. nematode recognition methods as claimed in claim 15, it is characterised in that control three-dimensional motion mechanism to adjust the number The relative position of code microscope and the objective table, including：
    Adjust motion of the digit microscope in the plane that X-axis and Y-axis are formed；And
    Adjust motion of the objective table on Z axis.
  17. 17. nematode recognition methods as claimed in claim 15, it is characterised in that control three-dimensional motion mechanism to adjust the number The relative position of code microscope and the objective table, including：Adjust three-dimensional of the digit microscope in X-axis, Y-axis and Z axis Motion.
  18. 18. nematode recognition methods as claimed in claim 11, it is characterised in that the smoothing processing uses medium filtering.
  19. 19. nematode recognition methods as claimed in claim 11, it is characterised in that believe the nematode image after smoothing processing Breath carries out self-adaption thresholding processing and contours extract, including：
    Centered on each pixel, the pixel in around it one square area delimited is averaged to determine this point The adaptive threshold at place；
    The pixel placement that will be above the adaptive threshold is 0, and the pixel placement that will be less than the adaptive threshold is 255；
    After the result of adaptive threshold is obtained, all white portions are found by extracting in image outmost outline The pixel on border, calculate the area that each outline is included and find out the profile that those areas exceed predefined size, delete not It is the white portion no more than predefined size of nematode, the white portion that area is exceeded into predefined size hooks as target area Rectangular bounding box and the circumscribed circle of these target areas are drawn to mark their position in the picture.
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