CN110066724A - Microculture real-time monitoring device and detection method - Google Patents
Microculture real-time monitoring device and detection method Download PDFInfo
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- CN110066724A CN110066724A CN201910483656.6A CN201910483656A CN110066724A CN 110066724 A CN110066724 A CN 110066724A CN 201910483656 A CN201910483656 A CN 201910483656A CN 110066724 A CN110066724 A CN 110066724A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/48—Holding appliances; Racks; Supports
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
Abstract
The present invention relates to microculture real-time monitoring device and detection methods, monitoring device includes culture cabinet, setting is in the intracorporal storage device of incubator, imaging device, operating device, completion sample storing unit and control system, detection method is that the culture dish in the storage device is continuously moved to the imaging device by the operating device, after imaging device obtains the culture dish image information, statistics compares, and forms testing result；It is moved back in the storage device again after imaging.The beneficial effects of the present invention are: realizing the environment of constant temperature and humidity in closed culture space, it will not influence Spawn incubation process while detection, it is continuously quickly continual to be detected, shorten Micro biological Tests duration, it can also record and save the details in microorganism growth process, the function of tracing to the source of microculture and detection is provided, reduces erroneous judgement risk to the maximum extent.
Description
Technical field
The invention belongs to microculture detection fields, more particularly, to a kind of microculture real-time monitoring device and inspection
Survey method.
Background technique
In research and development, production and the quality control in the fields such as food, cosmetics, drug, clinic, environment, it is often necessary to right
The growth metabolism of microorganism is observed and is studied, its growth rhythm is grasped, and analyzes its typicalness biobehavioral and feature.By
It is minimum in microorganism individual, it needs to be analyzed under amplification cultivation to microscope or naked eyes visible state, need by one day very
To culture in more than ten days, cause culture and observation analysis workload very big.The technology of microorganism scientific research at present and examination criteria are main
It carries out carrying out by the way of manual analysis after fixed incubation time, to the details in microorganism growth process, such as micro- life
Color change, bacterium colony size, colonial morphology, the speed of growth, sprawling or situation of movement, bacterium colony overlapping, bacterium in object growth course
It falls mutually inhibition, interferon secretion, data effectively cannot be obtained to situations such as erosion of culture medium, thus lack to micro- life
Object depth enters a possibility that research and analysis.
Time-consuming for artificial observation and record microculture outcome procedure, and workload is huge, the standard of culture and testing result
True property and stability dependency cause result standardization insufficient, note in factors such as operator Personal Skills, experience, mood, sense of responsibility
The record quality of data is not high, and culture and detection process can not accomplish that whole process can trace to the source；Can also there be erroneous judgement risk.The growth of microorganism
Slow device, logarithmic growth phase, stabilizer and decline phase can be roughly classified into.In different growth phases, the apparent performance of microorganism
It is different.If any microorganism representative configuration be generally present in stationary phase, arrived decline phase because there is changing for form and color
Become, leads to missing inspection；If any microorganism can occur to spread and migrate during the growth process, or even the entire culture dish of covering causes
Other bacterium colonies are covered completely；As the standard regulation mycotic culture time be 5 days after observe, spore can be generated by growing fast mould,
And two generations even three generations's bacterium colony out is sprouted, it is completely inconsistent with mould contamination actual conditions in sample；Microorganism also exists
Mutually inhibit the phenomenon that and kill, causes to occur different microorganisms population in microculture the feelings substituted occur in different time
Condition.Such phenomenon, cannot such as record in real time microorganism growth concrete condition, the observation result of large span timing node with
Actual conditions may not be inconsistent completely, lead to scientific research and the serious erroneous judgement of testing result.As job requirement is goed deep into and is detected in research
Increase, the workload of microorganism testing sharply increases.As required to use cascade sampling scheme in the work of current tainted food, compared with
More times were increased in simple sample scheme works amount in the past.By manual type, there are poles merely for a large amount of data record and analysis
It is big difficult, not only increase fallibility, it is also difficult to provide corresponding examining report.
The patent of Patent No. CN108192814 discloses a kind of method and apparatus counted for bacterium colony, and the patent is
Notice the importance that dynamic detection is carried out to microorganism growth process, but only in accordance with the mode of bacterium colony size to judge bacterium
Presence or absence is fallen, and counts the quantity of bacterium in growth course, but cannot to a large amount of typicalness information in bacteria growth process
It is analyzed and is grasped.And the counting is only taken pictures by the periodicity to fixed position culture dish and colony counts judge,
Cause culture dish capacity small, it is bulky, it is high to temperature control and space requirement.In addition one can only be once detected in the program
Culture dish will lead to take long time when culture dish quantity is larger, its corresponding detection cycle can also extend.
Summary of the invention
In order to solve the above technical problems, the present invention provides a kind of microculture real-time monitoring device and detection method, lead to
It crosses to microorganism in sequence culture medium ware culture, takes pictures by predetermined period, using uniqueness coding as crucial retrospect condition,
All electronic photos in microorganism growth process are stored, and by analysis system, any time point microorganism photo can be carried out
Manual analysis, or according to pre-set movement, electronic photo is automatically analyzed, related analysis report sum number is exported automatically
According to for studying and detecting application.
The technical solution adopted by the present invention is that: microculture real-time monitoring device, including culture cabinet, setting are being cultivated
Case intracorporal storage device, operating device, completes sample storing unit and control system, control system connection storage at imaging device
Device, imaging device and operating device, imaging device are connected with analytical equipment, and storage device and completion sample storing unit are set to
On the intracorporal culture table top of incubator, operating device is arranged in the operating table surface under culture table top.
Preferably, operating device includes the operating arm being separately positioned in operating table surface, takes ware bracket, send ware bracket and returns
It admits a child into a nursery school frame, operating arm is rotatably arranged, and takes ware bracket, and send ware bracket and recycling bracket to operating arm center of rotation apart from identical,
Ware bracket is taken, send ware bracket and recycling bracket that can move up and down.
Preferably, operating device further includes fetching and delivering sample isolation board, fetches and delivers sample isolation board and is provided projectingly for culture table top extension
Plate face above operating table surface fetches and delivers sample isolation board equipped with sampling hole and sample presentation hole, and sampling hole is arranged on taking ware bracket
Side, sample presentation hole are arranged above sample presentation bracket；
Preferably, sample presentation hole both sides are symmetrically arranged with sample presentation movement pawl, and sample presentation moves pawl rear end and rotates axis connection by sample presentation
On fetching and delivering sample isolation board, sample presentation movement pawl front end is reached in sample presentation hole；
Preferably, sample presentation rotation axis is equipped with reset spring；
Preferably, the bending of sample presentation movement pawl leading portion is downward；
Preferably, sampling bore edges, which are equipped with, takes ware baffle, and sample presentation bore edges are equipped with and send ware baffle；
Preferably, operating arm includes integrally formed lever arm and limit handgrip, and limit handgrip is annular or semi-circular；
Preferably, operating arm includes one or more.
Preferably, storage device is rotatable mouse cage, and basket support is equipped in mouse cage, can stacked arrangement culture dish, sampling
Hole and sample presentation hole respectively correspond two adjacent basket supports；
Preferably, culture dish is equipped with identification (RFID) tag；
Preferably, basket support is 2-16, and basket support is uniformly distributed in a ring.
Preferably, completing sample storing unit is recycling isolation board, and recycling isolation board is that culture table top extension is provided projectingly
Plate face above operating table surface, recycling isolation board are equipped with recovery holes, and recovery holes setting is above recycling bracket；
Preferably, recovery holes both sides are symmetrically arranged with recycling movement pawl, and recycling movement pawl rear end passes through recycling rotation axis connection
On recycling isolation board, recycling movement pawl front end is reached in recovery holes；
Preferably, recycling rotation axis is equipped with reset spring；
Preferably, the bending of recycling movement pawl leading portion is downward；
Preferably, recycling bore edges are equipped with recycling baffle.
Preferably, imaging device includes camera and platform of taking pictures, and platform of taking pictures concordantly is arranged in operating table surface, is taken a picture
The corresponding platform of taking pictures of machine camera lens, camera linking parsing device；
Preferably, imaging device further includes lighting device, the corresponding platform of taking pictures of lighting device.
Preferably, incubator includes culture table top, operating table surface and seal bootr, and seal bootr covers culture table top and station
Face forms closed culture space；
Preferably, seal bootr is transparent material；
Preferably, temperature control equipment and humidity conditioner, temperature control equipment and humid control are equipped in seal bootr
Device connects control system.
Using the microculture real-time detection method of microculture real-time monitoring device, continuously not by operating device
Disconnected is moved to the culture dish in storage device imaging device, and after imaging device obtains culture dish image information, statistics is compared
Testing result is formed, is moved back in storage device again after sampling；
Preferably, testing result meets or exceeds preset bounds, and culture dish is transferred to by operating device completes sample storage dress
It sets.
Preferably, the specific steps are as follows:
A culture dish in storage device is moved to imaging device by operating device by step 1；
Step 2 imaging device, which takes pictures to culture dish, obtains image information；
Step 3 image information generates bacterium colony information under the action of analytical equipment；
Culture dish is moved back in storage device by step 4 operating device, return step one；
In step 3, after bacterium colony information meets or exceeds preset bounds, implementation steps five are as follows:
Culture dish is moved to and completes sample storing unit, return step one by step 5 operating device.
Preferably, presupposed information includes independent bacterium colony size, colony colour, inhibition zone size, one of incubation time
Or it is a variety of,
Preferably, it is 25-60 DEG C that temperature is kept in incubator, humidity 10-85%；
Preferably, incubator, which is equipped with, picks and places mouth, can be put into storage device new in continual detection process
Culture dish will be completed the culture dish in sample storing unit and be taken out；
Preferably, when in operating device including multiple rotating arms, step 1, step 2, step 4 or step 5 are sequentially same
Step carries out.
Preferably, step 3 specific operation process is as follows:
The identification (RFID) tag that step A identification culture dish is equipped with determines culture dish number information, and by identification (RFID) tag to training
Support ware framing；
Step B measures bacterium colony information；
Step C combines the culture dish to have information and generates bacterium colony variation report；
Preferably, bacterium colony information includes bacterium colony size, colony colour, inhibition zone size, colony shape, bacterium colony in step B
One of quantity is a variety of；
Preferably, bacterium colony variation report includes growth curve information, strain conclusion information and judges in information in step C
It is one or more.
The advantages and positive effects of the present invention are:
1 is integrated in the microculture in microorganism detection, Data Detection statistical function in one device, can not stop
Microculture situation is continuously cultivated and monitored to machine；Culture can not be only counted according to the difference of Detection task content
The number of bacterium colony on ware, moreover it is possible to according to the calculation formula of industry regulation defined, final detection knot be calculated according to extension rate
Fruit quickly forms examining report；
2 this method can also record and save the details in microorganism growth process, in microorganism growth process
Color change, bacterium colony size, colonial morphology, the speed of growth, sprawling or situation of movement, bacterium colony overlapping, bacterium colony mutually inhibits, do
Disturb element secretion, data effectively cannot be obtained to situations such as erosion of culture medium, cause to lose to microorganism further investigation and
A possibility that analysis；
3 present invention can greatly mitigate the working strength of operator, improve the accuracy of detection, and provide micro- life
The function of tracing to the source of object culture and detection reduces erroneous judgement risk to greatest extent；
The environment of constant temperature and humidity is realized in 4 closed culture spaces, detection while will not influence Spawn incubation process, make
It is more stable and reliable that data must be cultivated；
5 continuously quickly continual can be detected, and shorten Micro biological Tests duration, data volume is bigger more quasi-
Really, scientific research demand can preferably be assisted.
Detailed description of the invention
Fig. 1 is one embodiment of the present of invention structural schematic diagram；
Fig. 2 is one embodiment of the invention operating table surface structural schematic diagram；
Fig. 3 is that one embodiment of the invention fetches and delivers sample partition plate structure schematic diagram；
Fig. 4 is that one embodiment of the invention fetches and delivers sample isolation board using structural schematic diagram；
Fig. 5 is one embodiment of the invention sample presentation movement pawl structural schematic diagram.
In figure:
1, cabinet 2, storage device 21, basket support are cultivated
3, sample storing unit 31, recycling isolation board 4, operating device are completed
41, arm 42 is operated, ware bracket 43 is taken, send ware bracket
44, bracket 5, imaging device 51, platform of taking pictures are recycled
6, sample isolation board 61, sampling hole 62, sample presentation hole are fetched and delivered
63, it takes ware baffle 64, send ware baffle 65, sample presentation movement pawl
66, sample presentation rotation axis
Specific embodiment
It explains with reference to the accompanying drawing to one embodiment of the present of invention.
As shown in Figure 1, microculture real-time monitoring device, including culture cabinet 1, depositing in culture cabinet 1 is set
Storage device 2, imaging device 5, operating device 4, completion sample storing unit 3 and control system, control system connection storage device 2,
Imaging device 5 and operating device 4, imaging device 5 are connected with analytical equipment.
Cultivating cabinet 1 includes culture table top, operating table surface and seal bootr, and seal bootr covers culture table top and operating table surface shape
At closed culture space, it is equipped with temperature control equipment and humidity conditioner in seal bootr, provides adaptation for microorganism growth
Stable growing environment, seal bootr are transparent material, facilitate situation in observation incubator；Temperature control equipment and humid control dress
Connection control system is set, temperature and humidity in incubator is adjusted by control system, wherein temperature control equipment and humid control
Device can be used the prior art that existing common microbial cultivation device uses, for example, reaction real time status temperature sensor and
Humidity sensor adjusts the heater of temperature and humidity, radiator, humidifier, the combination such as air circulating device.It is protected in incubator
Holding temperature is 25-60 DEG C, humidity 10-85%, and it is suitable that guarantee can also provide during detecting to each culture dish
Culture environment.Culture table top sink a part of region formation operation table top, storage device 2 and completes sample storing unit 3 and is set to
It cultivates on the culture table top in cabinet 1, operating device 4 is arranged in the operating table surface under culture table top.
Storage device 2 for storing microbial culture dish, for can horizontal direction circular-rotation mouse cage, mouse cage annular is equal
It is even to be equipped with basket support 21, each basket support 21 can stacked arrangement culture dish, each basket support 21 is made of the fixed link of circumferential arrangement, trains
It supports ware stacking to be placed in basket support 21, mouse cage driving device drives mouse cage rotation, and basket support 21 is driven to rotate, and basket support 21 rotates same
When drive inside culture dish rotate together, mouse cage driving device can be drivable rotation axis common in the art；
Basket support 21 can be 2-16, be evenly distributed in turning circumference, the distance of each basket support 21 to center of rotation is identical.Training
It supports ware and is equipped with identification (RFID) tag, such as bar code, two dimensional code or a string of numbers being made of text, letter and number, the label
Main application is first is that identify culture dish, second is that culture dish framing is used for, in order to which the different time of the same culture dish is clapped
The photo taken the photograph, which is done, subtracts shadow, to get rid of the influence that dust in culture dish, particle or spot etc. count bacterium colony.
As shown in Fig. 2, operating device 4 includes the operating arm 41 being separately positioned in operating table surface, ware bracket 42 is taken, ware is sent
Bracket 43 and recycling bracket 44, operating arm 41 are rotatably arranged, and take ware bracket 42, send ware bracket 43 and recycling bracket 44 to operating
41 center of rotation of arm takes ware bracket 42 apart from identical, send ware bracket 43 and recycling bracket 44 that can move up and down.Take ware bracket 42
It is made of bracket support and driving device, bracket support top is circular supporting plate, and lower part is the pressure pin being fixedly connected, and is connected under drawbar
There is driving device that bracket support is enabled to move up and down, operating table surface is equipped with and the matched round cavity of bracket support, bracket support
It is moved up and down in round cavity, while culture dish being driven to move up and down, bracket support extreme lower position is concordant with operating table surface；It send
Ware bracket 43 and recycling bracket 44 are identical as 42 structure of ware bracket is taken, and are the structures that can be moved up and down, can will be moved into
Culture dish in bracket support is held up to above culture table top, by moving the cooperation of pawl 65 or recycling movement pawl with sample presentation, is realized
The pick-and-place of culture dish.Wherein bracket support upper surface is concordant with operating table surface, and culture dish is facilitated to move left and right, and wherein driving device can
Think common drive, be connected to the control system, such as uses the cylinder connecting with control system.
Operating device 4 further includes fetching and delivering sample isolation board 6, as shown in Fig. 3 Fig. 4, fetches and delivers sample isolation board 6 and extends for culture table top
The plate face being provided projectingly above operating table surface fetches and delivers sample isolation board 6 equipped with sampling hole 61 and sample presentation hole 62, and sampling hole 61 is set
It sets and is taking above ware bracket 42, sample presentation hole 62 is arranged above sample presentation bracket, and 61 edge of sampling hole is equipped with and takes ware baffle 63, takes
Ware baffle 63 is curved or bending sheet metal, takes 63 leading portion of ware baffle prominent, can touch culture dish side wall, trains when filling with
The mouse cage for supporting ware turns at sampling hole 61, and individual culture dish outstanding trim locations under the action of taking ware baffle 63 make it
Ware bracket 42 can be taken below face, facilitate sampling；62 both sides of sample presentation hole are symmetrically arranged with sample presentation movement pawl 65, as shown in figure 5,
Sample presentation movement 65 rear end of pawl is connected to and is fetched and delivered on sample isolation board 6 by sample presentation rotation axis 66, enable sample presentation movement pawl 65 around
About 66 sample presentation rotation axis is stirred, and sample presentation movement 65 front end of pawl reaches in sample presentation hole 62, is used to support in sample presentation hole 62
The culture dish of top can be equipped with reset spring in sample presentation rotation axis 66, make to increase the rebound rate of sample presentation movement pawl 65
Obtaining sample presentation movement pawl 65 can quickly spring back；Sample presentation movement pawl 65 is U-shaped, and sample presentation moves the bending of 65 leading portion of pawl downwards in wedge angle
It is hook-shaped, it also avoids increasing the distance that sample presentation movement pawl 65 carries culture dish while increasing sample presentation movement 65 length of pawl, improves work
Make efficiency, guarantees culture dish stability after sample presentation；In order to guarantee that the culture dish of sample presentation is put neatly, 62 edge of sample presentation hole is equipped with and send
Ware baffle 64 send ware baffle 64 can be in sample presentation when culture dish is out of alignment for the sheet metal of arc or bending shape
Process increases the stability of sample presentation accumulative culture ware by sending the effect of ware baffle 64 to be finely adjusted position.During taking ware,
One culture dish height of decline after taking ware can be preset, is rotated further by mouse cage, or at the culture next culture dish height of table top
Position sensor is set, after sensing that the last one culture dish reaches this position, stops and takes ware bracket 42, rotates mouse cage.
It completes sample storing unit 3 to be used to place the culture dish for completing culture detection, pre-sets culture to control system
The upper limit of time or indices, can be by operating device 4 by culture dish when culture dish meets or exceeds corresponding index
It moves on to and completes sample storing unit 3；Completing sample recyclable device is specially to recycle isolation board 31, and recycling isolation board 31 is that culture table top prolongs
The plate face being provided projectingly above operating table surface is stretched, recycling isolation board 31 is equipped with recovery holes, recovery holes and 62 structure of sample presentation hole
Similar, culture dish passes through recovery holes under the action of recycling bracket 44 and reaches above recycling isolation board 31, moves pawl by recycling
It blocks, is retained on recycling isolation board 31.Recovery holes both sides are symmetrically arranged with recycling movement pawl, and recycling movement pawl rear end passes through recycling
Rotation axis is connected on recycling isolation board 31, and recycling movement pawl can be rotated around rotation axis, and recycling movement pawl front end reaches back
In batter, the culture dish of recycling is moved up under the action of recycling bracket 44 across recovery holes, and recycling movement pawl front end rides over culture
As outside is opened upwards for the rising of culture dish on ware, when culture dish rises to a certain extent, recycling movement pawl front end is taken not
Firmly culture dish is fallen after rise under gravity, at this time puts down culture dish, and culture dish can be stuck in the front end of recycling movement pawl, from
And complete the operating process of recycling culture dish；In order to increase the rebound rate of recycling movement pawl, can be equipped in recycling rotation axis
Reset spring enables recycling movement pawl quickly to spring back；Recycling movement pawl leading portion bending increases back downwards in the hook-shaped of wedge angle
It also avoids increasing the distance that recycling movement pawl carries culture dish while receiving movement pawl length, improves working efficiency, guarantee recycling
Culture dish stability afterwards.In order to guarantee that the culture dish of recycling is put neatly, recycling bore edges are equipped with recycling baffle, and recycling baffle is
The sheet metal of arc or bending shape can pass through the work of recycling baffle in removal process when culture dish is out of alignment
It is finely adjusted with to position, increases the stability of recycling accumulative culture ware.62 two sides of sample presentation hole are respectively equipped with mounting hole, and sample presentation turns
Moving axis 66 is arranged in mounting hole.
Operating arm 41 includes integrally formed lever arm and limit handgrip, and lever arm bottom is equipped with rotation drive device, and rotation is driven
Dynamic device is the rotation axis or rotate gear with driving motor common in the art, rotation drive device and the control
System connection, actuated lever arm rotate clockwise or counterclockwise, and limit handgrip is annular or semi-circular, and it is flat to be able to drive culture dish
It moves；Operating arm 41 includes one or more, and culture dish can be moved realization detection process in sequence, be by an operating arm 41
Detection speed is improved, multiple operating arms 41 can be set, successively carries out detecting according to clockwise or counterclockwise
At this moment journey can set gradually sample presentation hole 62, sampling hole 61,51 He of platform of taking pictures according to clockwise or counterclockwise
Recovery holes, wherein sample presentation hole 62 and sampling hole 61 are corresponding in turn to two adjacent basket supports 21.
Imaging device 5 includes camera and platform 51 of taking pictures, and platform 51 of taking pictures concordantly is arranged in operating table surface, camera
The corresponding platform 51 of taking pictures of camera lens, camera linking parsing device, camera can be set on the side wall of culture cabinet 1, imaging
Device 5 further includes lighting device, is similarly disposed on culture 1 side wall of cabinet, corresponding platform 51 of taking pictures, when take pictures insufficient light when
For illuminating light filling；Camera is connected with analytical equipment, is specifically as follows computer, and camera passes through Ethernet interface and computer
It is connected, the information for obtaining culture dish is analyzed by computer, and report is formed by further analysis.
Control system can be control circuit board, and control circuit board controls each driving device and realizes automatic detection function
Can, outside is connect by input and output interfaces with computer, to realize control function or set-up function.
In use, multiple culture dishes are marked, it is placed into the basket support 21 of culture cabinet 1, when basket support 21 turns to
When at sampling hole 61, ware bracket 42 is taken to rise, bracket support is concordant with culture table top, and the culture dish of lowermost end connects with ware bracket 42 is taken
Touching, takes ware bracket 42 to move down the height of a culture dish, and basket support 21 turns round the distance of half of culture dish, penultimate training
It supports ware half of culture dish under the action of 21 fixed link of basket support and culture table top to turn to outside 61 range of sampling hole, takes ware bracket 42
It moves down, the culture dish of least significant end is driven to drop in operating table surface, remaining culture dish is also retained in basket support 21；Work as training
Feeding ware drop to it is concordant with operating table surface, operating arm 41 rotate, drive drop into limit handle in culture dish be moved to take pictures it is flat
Platform 51 is analyzed after camera sampling by computer, and operating arm 41 drives culture dish to turn to again and send at ware bracket 43, lead to
It crosses and the cooperation for lifting and sending ware movement pawl of ware bracket 43 is sent to send culture dish back to basket support 21, ware bracket 43 is sent to lift on culture dish
It moves, after passing through sample presentation hole 62, sample presentation movement pawl 65 is ridden on culture dish, and when sending ware bracket 43 to continue to move up, sample presentation moves pawl 65
Spring back and fall after being detached from culture dish side wall, sample presentation moves pawl 65 and drops back into horizontal, send ware bracket 43 to move down, culture dish, which is stuck in, to be sent
Sample moves on pawl 65, avoids falling, ware bracket 43 is sent to continue to move down, and waits and transports next culture dish；In one basket support 21
For culture dish all after detection, mouse cage rotation detects the culture dish in new basket support 21, keeps having in mouse cage a basket support 21 be
Sky, for storing the culture dish detected, such structure sequentially continuously continual can be detected, and incubator is equipped with and takes
Mouth is put, new culture dish can be put into storage device 2 in continual detection process, will be completed in sample storing unit 3
Culture dish take out, when there is new culture dish to need to be added, only need to open seal bootr addition, without stop detection.
Using the microculture real-time detection method of microculture real-time monitoring device, continuously not by operating device
The culture dish in storage device disconnectedly is moved to imaging device, after imaging device obtains culture dish image information, statistics is compared
Testing result is formed, is moved back in storage device again after sampling；Testing result meets or exceeds preset bounds, and operating device will be trained
Feeding ware, which is transferred to, completes sample storing unit.
Specific step is as follows:
A culture dish in storage device is moved to imaging device by operating device by step 1；
Step 2 imaging device, which takes pictures to culture dish, obtains image information；
Step 3 image information generates bacterium colony information under the action of analytical equipment；
Culture dish is moved back in storage device by step 4 operating device, return step one；
In step 3, after bacterium colony information meets or exceeds preset bounds, such as independent bacterium colony size, colony colour are antibacterial
Circle size, the conditions such as incubation time, implementation steps five are as follows:
Culture dish is moved to and completes sample storing unit, return step one by step 5 operating device.
Analytic process specific operation process is as follows in step 3:
The identification (RFID) tag that step A identification culture dish is equipped with determines culture dish number information, and by identification (RFID) tag to training
Support ware framing；；
Step B measures bacterium colony information such as bacterium colony size, colony colour, inhibition zone size, colony shape, colony counts etc.；
Step C combines the culture dish to have information and generates bacterium colony variation report, it may include growth curve information, strain conclusion
Information and judge information.
When including multiple rotating arms in operating device, step 1, step 2, sequentially synchronization is carried out for step 4 or step 5,
It can accelerate to detect speed, improve detection efficiency.
Concrete operation step can be as follows,
1, identify the method for culture dish: identification (RFID) tag is posted in culture dish bottom (can be bar code, two dimensional code or by text
A string of numbers of word, letter and number composition).The label main application, first is that identification culture dish, second is that being used for culture dish image
Positioning, subtract shadow in order to which the photo that the different time of the same culture dish takes is done, thus get rid of dust in culture dish,
The influence that grain or spot etc. count bacterium colony.
2, realize the method for bacterium colony dimensional measurement: bacterium colony size is to convert to obtain by pixel.In system program, if
It is equipped with calibrating length function.The basic skills of calibrating length is the article (such as culture dish diameter) for shooting known length, software
System calculates pixel-length conversion coefficient K (mm/ pixel), the i.e. length of the actual object of each pixel representative of image automatically
Degree.When needing to measure bacterium colony size, the pixel number in some direction of the bacterium colony identified multiplied by COEFFICIENT K, can be obtained
Bacterium colony physical length.
3, the realization of color analysis function: different bacterium colonies has different colors, colored digital used by the present apparatus
The available colouring information to bacterium colony of camera is aided with other parameters (such as shape) according to color, and computer software can identify
Bacterium colony type out.
4, inhibition zone measurement function: when studying inhibiting effect of certain drug to microorganism, drug drip to be measured can be arrived
In culture dish, drug is spread in agar plate, is suppressed surrounding bacterial growth to forming transparent circle, i.e., antibacterial
Circle, can be determined that the inhibitory potency of drug to be measured according to inhibition zone size.This system computer program contains inhibition zone measurement function,
I.e. by image recognition, the size of inhibition zone is measured, to calculate the inhibitory potency of tested drug automatically.
5, bacterium colony clustering, colony shape are fitted function: the growth of every kind of bacterium colony has reapective features, is mainly reflected in bacterium
It falls in shape.Present apparatus software systems, by being continuously shot the growth course of bacterium colony in the same culture dish, using a variety of images
Recognition methods (including cluster algorithm), in conjunction with known colonial morphology, to identify bacterium colony type.
6, bacterium colony number growth curve: each culture dish can be photographed once at regular intervals (such as 30,60 minutes),
Bacterium colony number on every photo can be counted automatically once by software systems again, thus can be the bacterium colony of each culture dish
Number, which changes with time, draws curve, i.e. bacterium colony number growth curve.It can be seen that, some bacterium colonies are being grown into most from curve
After multiple numbers, because two adjacent bacterium colonies may be fused into a bacterium colony, total plate count is caused to decline.It, can be with by the curve
The most bacterium colony number of culture dish is read, to reduce counting error.
The course of work of this example:
1) culture dish transport and process of taking pictures
After booting, under the control of the computer, initial position is found in cultivation region mouse cage rotational positioning.Operator will prepare
Good culture dish overlays in each basket support, keeps an empty basket support；Control temperature unit work, guarantees that temperature in chassis reaches setting
Temperature value (such as 37 DEG C)；
Operating arm rotates to and takes ware position, takes ware bracket uplink, touches in basket support downlink one after the last one culture dish
A culture dish height, basket support revolution, fixed residue culture dish take ware bracket that the last one culture dish is driven to be moved downward to and grasp
It is concordant to make table top, culture dish is operated arm and catches, and operating arm turns to shooting area, cameras work, and bat takes a culture dish
Photo, photo are transmitted in computer；The dynamic culture dish of operating armband, which rotates together, send ware position, send ware bracket uplink, will cultivate
Ware is sent into empty basket support, and ware bracket is sent to come downwards to minimum point；
In one basket support after detection, mouse cage rotation makes the basket support with culture dish turn to fetch bit and sets, and repetition takes training
Ware is supported, takes pictures, send basket support back to.It repeats the above process, until the plate in basket support is all taken pictures one time.
2) bacterium colony detection, counting process
The number and bacterium colony of culture dish were photographed on each photo, system configuration microorganism image analysis system can be right
Each microbial culture dish photo carries out detailed image analysis, wherein including but not limited to identification culture dish number, bacterium colony
Dimensional measurement function, color analysis function, clustering function, condition count function, inhibition zone measurement function, colony shape are quasi-
Close function, draw bacterium colony number growth curve function etc..
3) culture dish is sent out
Stationary phase is come into when determining that a certain culture dish bacterium colony is grown, or the incubation time (such as 24 hours) set is
Through arriving, which, which is operated arm and is sent to, completes area.Bracket uplink is recycled, is ejected and is completed on sample isolation board.
4) examining report generates
The algorithm of various microorganism monitorings and detection, calculating and report generation, Ke Yigen are stored in computer software system
According to monitoring and testing result, final detection result is calculated according to corresponding algorithm automatically, and print report.
5) detection process is traced to the source
The in store previous whole testing results of database in computer, including the photograph of each sample shot in the process
Piece.Operator can inquire at any time, recall these photos and result.Detect the weight such as source, operator's name of sample
Information is wanted also to save in the database.
Embodiment 1:
Microculture real-time monitoring device, including culture cabinet, setting is in the intracorporal storage device of incubator, imaging dress
It sets, operating device, complete sample storing unit and control system, control system connects storage device, imaging device and operating device,
Imaging device is connected with analytical equipment, and storage device and completion sample storing unit are set on the intracorporal culture table top of incubator,
Operating device is arranged in the operating table surface under culture table top.
Storage device is rotatable mouse cage, and basket support is equipped in mouse cage, and basket support is 16, and basket support annular is uniformly distributed, energy
Enough stacked arrangement culture dishes, culture dish are equipped with identification (RFID) tag, and sampling hole and sample presentation hole respectively correspond two adjacent basket supports.
Operating device includes the operating arm being separately positioned in operating table surface, takes ware bracket, send ware bracket and recycling bracket,
Operating arm is rotatably arranged, and takes ware bracket, send ware bracket and recycling bracket to arm center of rotation is operated apart from identical, takes ware support
Frame send ware bracket and recycling bracket that can move up and down；Further include fetching and delivering sample isolation board, fetches and delivers sample isolation board and prolong for culture table top
The plate face being provided projectingly above operating table surface is stretched, fetches and delivers sample isolation board equipped with sampling hole and sample presentation hole, sampling hole setting exists
It takes above ware bracket, sample presentation hole is arranged above sample presentation bracket；Sample presentation hole both sides are symmetrically arranged with sample presentation movement pawl, and sample presentation moves pawl
Rear end is connected to by sample presentation rotation axis and is fetched and delivered on sample isolation board, and sample presentation rotation axis is equipped with reset spring, before sample presentation moves pawl
End reaches in sample presentation hole, and leading portion bending is downward；Sampling bore edges, which are equipped with, takes ware baffle, and sample presentation bore edges are equipped with and send ware baffle；
Operating arm includes integrally formed lever arm and limit handgrip, and limit handgrip is semi-circular.
Completing sample storing unit is recycling isolation board, and recycling isolation board is that culture table top extension is provided projectingly in operating table surface
The plate face of top, recycling isolation board are equipped with recovery holes, and above recycling bracket, recovery holes both sides are symmetrically arranged with for recovery holes setting
Recycling movement pawl, recycling movement pawl rear end are connected on recycling isolation board by recycling rotation axis, and recycling rotation axis is equipped with multiple
Position spring, downwards, recycling movement pawl front end reaches in recovery holes the bending of recycling movement pawl leading portion, and recycling bore edges are equipped with recycling
Baffle.
Imaging device includes camera and platform of taking pictures, and platform of taking pictures concordantly is arranged in operating table surface, camera gun
Corresponding platform of taking pictures, camera linking parsing device；Imaging device further includes lighting device, the corresponding platform of taking pictures of lighting device.
Incubator includes culture table top, operating table surface and seal bootr, and seal bootr is transparent material, and seal bootr covers culture table top and operation
Table top forms closed culture space, is equipped with temperature control equipment and humidity conditioner in seal bootr, temperature control equipment and
Humidity conditioner connects control system.
One embodiment of the present invention has been described in detail above, but the content is only preferable implementation of the invention
Example, should not be considered as limiting the scope of the invention.It is all according to all the changes and improvements made by the present patent application range
Deng should still be within the scope of the patent of the present invention.
Claims (10)
1. microculture real-time monitoring device, it is characterised in that: including cultivating cabinet, setting is deposited the incubator is intracorporal
Storage device, operating device, completes sample storing unit and control system at imaging device, and the control system connects the storage dress
It sets, the imaging device and the operating device, the imaging device is connected with analytical equipment, the storage device and described complete
It is set on the intracorporal culture table top of the incubator at sample storing unit, the operating device is arranged under the culture table top
Operating table surface on.
2. microculture real-time monitoring device according to claim 1, it is characterised in that: the operating device includes point
Operating arm in the operating table surface is not set, ware bracket is taken, send ware bracket and recycling bracket, the operating arm is rotatably set
It sets, it is described to take ware bracket, it is described to send ware bracket and the recycling bracket to the operating arm center of rotation apart from identical, it is described to take
Ware bracket, it is described to send ware bracket and the recycling bracket that move up and down.
3. microculture real-time monitoring device according to claim 2, it is characterised in that: the operating device further includes
Sample isolation board is fetched and delivered, the sample isolation board of fetching and delivering is that the culture table top extends the plate being provided projectingly above the operating table surface
Face, equipped with sampling hole and sample presentation hole, the sampling hole setting takes above ware bracket the sample isolation board of fetching and delivering described, described
Sample presentation hole is arranged above the sample presentation bracket；
Preferably, sample presentation hole both sides are symmetrically arranged with sample presentation movement pawl, and sample presentation movement pawl rear end passes through sample presentation rotation axis
It is fetched and delivered on sample isolation board described in being connected to, sample presentation movement pawl front end reaches in the sample presentation hole；
Preferably, the sample presentation rotation axis is equipped with reset spring；
Preferably, the sample presentation movement pawl leading portion bending is downward；
Preferably, the sampling bore edges, which are equipped with, takes ware baffle, and the sample presentation bore edges are equipped with and send ware baffle；
Preferably, the operating arm includes integrally formed lever arm and limit handgrip, and the limit handgrip is annular or semi-circular；
Preferably, the operating arm includes one or more.
4. microculture real-time monitoring device according to claim 3, it is characterised in that: the storage device is that can turn
Dynamic mouse cage is equipped with basket support in the mouse cage, can stacked arrangement culture dish, the sampling hole and the sample presentation hole respectively correspond
Two adjacent basket supports；
Preferably, the culture dish is equipped with identification (RFID) tag；
Preferably, the basket support is 2-16, and the basket support is uniformly distributed in a ring.
5. microculture real-time monitoring device according to claim 3, it is characterised in that: the completion sample storing unit
To recycle isolation board, the recycling isolation board is that the culture table top extends the plate being provided projectingly above the operating table surface
Face, the recycling isolation board are equipped with recovery holes, and the recovery holes are arranged above the recycling bracket；
Preferably, the recovery holes both sides are symmetrically arranged with recycling movement pawl, and the recycling movement pawl rear end passes through recycling rotation axis
It is connected on the recycling isolation board, recycling movement pawl front end reaches in the recovery holes；
Preferably, the recycling rotation axis is equipped with reset spring；
Preferably, the recycling movement pawl leading portion bending is downward；
Preferably, the recycling bore edges are equipped with recycling baffle.
6. microculture real-time monitoring device according to claim 2 or 3, it is characterised in that: the imaging device packet
It includes camera and platform of taking pictures, the platform of taking pictures concordantly is arranged in the operating table surface, the camera gun corresponds to institute
Platform of taking pictures is stated, the camera connects the analytical equipment；
Preferably, the imaging device further includes lighting device, platform of taking pictures described in the lighting device correspondence.
7. microculture real-time monitoring device according to claim 1 to 3, it is characterised in that: the incubator
Including the culture table top, the operating table surface and seal bootr, the seal bootr covers the culture table top and the station
Face forms closed culture space；
Preferably, the seal bootr is transparent material；
Preferably, temperature control equipment and humidity conditioner are equipped in the seal bootr, the temperature control equipment and described
Humidity conditioner connects the control system.
8. using the microculture real-time detection side of any microculture real-time monitoring device in claim 1-7
Method, it is characterised in that: by the operating device continuously by the culture dish in the storage device be moved to it is described at
As device, after imaging device obtains the culture dish image information, statistics compares and forms testing result, moves back to institute after sampling again
It states in storage device；
Preferably, the testing result meets or exceeds preset bounds, and the culture dish is transferred to described by the operating device
Complete sample storing unit.
9. microculture real-time detection method according to claim 8, it is characterised in that: specific step is as follows:
A culture dish in storage device is moved to imaging device by operating device by step 1；
Step 2 imaging device, which takes pictures to culture dish, obtains image information；
Step 3 image information generates bacterium colony information under the action of analytical equipment；
Culture dish is moved back in storage device by step 4 operating device, return step one；
In step 3, after bacterium colony information meets or exceeds preset bounds, implementation steps five are as follows:
Culture dish is moved to and completes sample storing unit, return step one by step 5 operating device.
Preferably, presupposed information includes independent bacterium colony size, colony colour, inhibition zone size, one of incubation time or more
Kind,
Preferably, it is 25-60 DEG C that temperature is kept in incubator, humidity 10-85%；
Preferably, the incubator, which is equipped with, picks and places mouth, can be put into Xiang Suoshu storage device in continual detection process
New culture dish takes out the culture dish in the completion sample storing unit；
Preferably, when in the operating device including multiple rotating arms, the step 1, step 2, step 4 or step 5 are suitable
Subsynchronous progress.
10. microculture real-time detection method according to claim 9, it is characterised in that: the step 3 is specifically grasped
It is as follows to make process:
The identification (RFID) tag that step A identification culture dish is equipped with determines culture dish number information, and by identification (RFID) tag to culture dish
Framing；
Step B measures bacterium colony information；
Step C combines the culture dish to have information and generates bacterium colony variation report；
Preferably, bacterium colony information includes bacterium colony size, colony colour, inhibition zone size, colony shape, bacterium colony in the step B
One of quantity is a variety of；
Preferably, bacterium colony variation report includes growth curve information, strain conclusion information and judges in information in the step C
It is one or more.
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