CN109321429A - A kind of automatical bacterium instrument and microbe to screen method - Google Patents
A kind of automatical bacterium instrument and microbe to screen method Download PDFInfo
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- CN109321429A CN109321429A CN201811140500.XA CN201811140500A CN109321429A CN 109321429 A CN109321429 A CN 109321429A CN 201811140500 A CN201811140500 A CN 201811140500A CN 109321429 A CN109321429 A CN 109321429A
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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
- C12M23/02—Form or structure of the vessel
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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
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
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Abstract
The invention discloses a kind of automatical bacterium instrument and microbe to screen methods, automatical bacterium instrument includes liquor removing workstation and nephelometer, nephelometer is mounted on liquor removing workstation, and the invention also discloses the microbe to screen method based on automatical bacterium instrument, this method is used for the acquisition of single bacterial strain.Liquor removing workstation combination nephelometer of the invention, compared to previous liquor removing workstation combination microplate reader, it is lower to measure influence of the environment for the growth of microorganism, it enables the invention to also can fast and accurately measure the growing state of determining microorganism in the case where anaerobic environment reduces exposure duration to the greatest extent with other；In addition, nephelometer can directly read the picking sequential file for generating liquor removing workstation in hole after turbidity, for liquor removing workstation picking microorganism.The present invention is based on the screening techniques for the acquisition for single bacterial strain that automatical bacterium instrument provides, and are screened by using automatical bacterium instrument, come out single strain isolation from complex sample, obtain the single bacterial strain obviously grown.
Description
Technical field
The present invention relates to a kind of automatical bacterium instrument and its microbe to screen method more particularly to a kind of high throughputs of microorganism
That cultivates and screen chooses bacterium instrument and using the method for choosing bacterium instrument screening microorganism.
Background technique
Liquor removing workstation is that one kind can carry out high-throughput liquid reagent transfer, diluted instrument automatically.It can be according to micropore
The setting of board type and significant hole location is automatically performed the work such as transfer, dilution and distribution.High pass of the equipment in microorganism
There is important value in amount culture and screening operation, each hole location of microwell plate is read by using instruments such as microplate reader
Then value determines microorganism growing state, then the hole that artificial setting needs to extract and shift according to the result of reading and threshold value
Position, thus considerably increases the working time.When culture have to anaerobic environment or other to reduce exposed to the greatest extent when
Between in the case where, microplate reader, which reads and then carries out shifting this process, may cause the destruction of culture.
Nephelometer can overcome this defect, and nephelometer is the device for measuring liquid turbidity, there is scattering type, transmitted light formula
With transmitted scattered light formula etc., it is referred to as optical profile type nephelometer.Its principle is, when light is irradiated on liquid level, incident intensity, transmitted light
By force, there are certain correlativities between the mutual ratio of scattered light intensity and water sample turbidity, pass through measurement transmitted light intensity, scattering
The ratio of light intensity and incident intensity or transmitted light intensity and scattered light intensity measures the turbidity of water sample.Optical profile type nephelometer has for real
Test room, it is also useful automatically continuously to be measured on site.
Microplate reader or nephelometer volume at present on the market is larger, and data streams read can not be direct by liquor removing workstation
It uses.It based on this, needs to design a kind of automatical bacterium instrument, in conjunction with liquor removing workstation and nephelometer, realizes efficient mask work,
Simultaneously anaerobic environment or other to reduce exposure duration to the greatest extent and quick and precisely can also efficiently screen microorganism.
Summary of the invention
In order to solve problem above, the object of the present invention is to provide a kind of automatical bacterium instrument and its microorganism high-pass to screen
Method, the equipment can culture rapidly and efficiently and screening microorganism, facilitate researcher to use.
In order to achieve the goal above, the present invention provides a kind of automatical bacterium instrument, comprising:
Liquor removing workstation, the liquor removing workstation include liquid suction module and Y-axis pallet, the liquid suction module peace
In the upper housing of liquor removing workstation, liquid suction module realizes that X-axis is mobile by X-axis driver drives, the Y-axis pallet peace
On the support board of liquor removing workstation bottom end, Y-axis pallet realizes that Y-axis is mobile by Y-axis driver drives, on the Y-axis pallet
Equipped with multiple areas Ban Wei, the area Ban Wei positioned at the lower left corner is hollow structure.The liquid suction module of the liquor removing workstation can be right
Liquid is aspirated, by realizing the transfer and dilution of liquid in X-axis movement.
Nephelometer, the nephelometer are mounted on the outside of the area Ban Wei in the lower left corner, and the nephelometer includes by stepper motor band
The dynamic U-shaped arm for realizing that Y-axis is mobile, Y-axis pallet are located between the upper lower wall bar of the U-shaped arm, divide on the upper lower wall bar of the U-shaped arm
It She You not transmitter and the receiver with transmitter cooperation.When work, the area Ban Wei in the lower left corner is placed with microwell plate, and microwell plate is
Transparent structure, for U-shaped arm in X-direction movement, every hole for reaching microwell plate, the light that transmitter issues is through microwell plate
It after liquid in hole, is received by the receiver, the signal received according to receiver, control system calculates the muddiness of liquid in hole
Degree, and then learn microorganism growing state inside each hole of microwell plate.
Control system, the control system include controller, the controller and liquor removing workstation and turbidimetric signal
Connection, controller are conventional single-chip microcontroller, are implanted into process control liquor removing workstation and turbidimetric operation；Specifically, for moving
Liquid work station, the liquid suction of control liquid suction module, control X-axis driver drives liquid suction module are moved in X-direction
Dynamic, control Y-axis driver drives Y-axis pallet is mobile in Y direction；For nephelometer, control transmitter emits light, according to connecing
The reception information of device is received, and calculates the turbidity of liquid in micropore, control stepper motor drives U-shaped arm in the mobile reading of Y direction
Take fluid information in micropore.
Further, the nephelometer further includes left plate, right side plate, the bottom plate, preceding connection for constituting nephelometer skeleton
Plate and rear connecting plate, the stepper motor and U-shaped arm are located in skeleton, hood are equipped on the upside of the skeleton, to intraskeletal
Member shades light processing, enables receiver successfully to receive the signal of transmitter sending, prevents ambient interference effect receiver
The light that transmitter issues is received, and then influences the judgement to liquid turbidity in micropore.
Further, being equipped with screw rod and gag lever post between the left plate and right side plate, the stepper motor passes through transmission
Gear connecting screw, the threaded hole that the screw rod passes through on the downside of U-shaped arm are threadedly coupled with U-shaped arm, and the gag lever post passes through U-shaped arm
The through-hole of downside.Rotary motion is transformed to move along a straight line by screw rod, movement accuracy is high, and has very big reduction of speed ratio, ties
Structure is simple, easily manufactured, stable working, noiseless, can transmit very big axial force；Gag lever post makes U-shaped arm can only be along screw rod
Length direction movement, prevent U-shaped arm as screw rod rotates, U-shaped arm made accurately to scan the microwell plate between U-shaped arm；Driving cog
Wheel is that reduction gearing combines, and the high-speed rotation of stepper motor is made to become slowly running for screw rod through transmission gear conversion.
Further, being equipped with initial level sensor close to left plate end on connecting plate after described, the bottom plate is equipped with
Control circuit board, right side plate are equipped with the USB interface of connection control circuit board.When not starting to read microwell plate, U-shaped arm is initial
Change returns at initial level sensor, receives the signal of transmitter without receiver under plate in no light and illumination, is changed by control system
It calculates, position the zero point of coloration and works to the last minute, be used for subsequent calculations illumination transmissivity；Turbidimetric control is installed in control circuit board
Circuit processed, control circuit control and receive the operation of device, transmitter and stepper motor, by the optical signal of receiver acquisition by processing
It is converted into the turbidity of liquid, control circuit simultaneously connect with control system signal, controlled by control system, and control system is according to returning
The liquid turbidity information returned can automatically produce picking file；USB interface is used for the connection of nephelometer and external equipment and leads to
News.
Further, the transmitter carries out transmitting light collimation using 600-605nm LED and lens group or has
The 650nm semiconductor laser of collimation lens, this is to be selected according to the detection of classical microorganism concn using OD600 and OD660；
The receiver is adopted using CdS photo resistance since the nephelometer local environment stray light being incorporated on liquor removing workstation is more
It uses photocell to be easy to be saturated by environment light as receiver, therefore uses CdS photo resistance, it is linear with preferable concentration signal
Feature.
Further, the liquid suction module includes multiple pipetting heads, the pipetting head is driven by sucker rod and is realized
The suction of liquid.The principle of liquid suction module is aspirated similar to syringe, is moved up and down by sucker rod, is made on pipetting head suction pallet
Liquid, pipetting head can also be mobile in vertical direction, and by pipetting head pumping liquid, pipetting head pumping liquid is by control system
Control, allows liquor removing workstation to carry out the transfer and dilution of liquid reagent, the high flux screening of convenient microbe.
Further, the support board is equipped with reject chute, reject chute is incline structure, and the liquid being finished after discarding is straight
It connects and is poured in discarded slot, by discarded slot discharge.
The present invention provides a kind of microbe to screen method of automatical bacterium instrument, this method is used for the acquisition of single bacterial strain,
The following steps are included:
Step S1 prepares primary sample, is diluted primary sample with the liquor removing workstation of automatical bacterium instrument, after dilution
Culture medium is added in primary sample, obtains media samples, is dispensed into media samples with the liquor removing workstation of automatical bacterium instrument
In the micropore of microwell plate.Here primary sample is complex sample, and primary sample contains multiple bacterial strains, the purpose of this method
It is to obtain the single bacterial strain of target in complex samples.
Step S2, by the media samples in microwell plate, constant temperature incubation is to object bacteria at a temperature of aimed strain growth
Strain growth.Aimed strain refers to needing the single bacterial strain that obtains, aimed strain the bacterial strain suitable growth at a temperature of cultivate
After a period of time, aimed strain growth；The suitable growth temperature and cultivated days of aimed strain are obtained by those skilled in the art
After knowing the title of the bacterial strain, the routine growth environment for inquiring the bacterial strain is determined.
Step S3, the media samples that culture is completed are read each on microwell plate with the nephelometer on automatical bacterium instrument
The turbidity value of culture medium in micropore, and generate picking file.
Step S4, automatical bacterium instrument set liquor removing workstation and choose bacterium minimum according to nephelometer testing result in picking file
OD value, liquor removing workstation carry out picking to the culture medium in the micropore for being greater than minimum OD value, i.e., selection microorganism has grown micro-
Culture medium in hole carries out picking, obtains single bacterial strain.
Further, nephelometer reads each on microwell plate in microbe to screen method and step S3 provided by the invention
In micropore culture medium turbidity value the following steps are included:
Microwell plate, is placed in the lower left corner position of Y-axis pallet by step S31, and turbidimetric U-shaped arm initialization returns to initial bit
At sensor, and read the signal of no light and illumination without plate when, it is determined as the zero point of transmission and work to the last minute, for calculating transmission
Rate；
Step S32, Y-axis pallet is mobile to nephelometer direction, when the first round for being moved to microwell plate is in U-shaped arm lower part
Light source right above after stop movement, then U-shaped arm according to the program of setting from initial position to distally gradually moving, every arrival
Controller reads the transmissivity in the hole after below one hole, and is converted into OD value；
Step S33 repeats reading of the step S31-S32 completion to entire microwell plate, and generates picking file by controller,
Controller is single-chip microcontroller.
Beneficial effects of the present invention:
Liquor removing workstation combination nephelometer of the invention measures ring compared to previous liquor removing workstation combination microplate reader
Influence of the border for the growth of microorganism is lower, enables the invention to reduce the feelings of exposure duration to the greatest extent with other in anaerobic environment
Under condition, the growing state of determining microorganism also can be fast and accurately measured；In addition, nephelometer can directly read life after turbidity in hole
At the picking sequential file of liquor removing workstation, for liquor removing workstation picking microorganism；
Liquor removing workstation provided by the invention can be shifted and be diluted to liquid reagent, and liquid suction module is to liquid
Body is aspirated, and X-axis driver drives liquid suction module is mobile in X-direction, and Y-axis driver drives Y-axis pallet is in Y-axis side
To movement, the pipetting head of liquid suction module can also be mobile in vertical direction, realize three-shaft linkage, to putting on Y-axis pallet
The microwell plate set carries out the transfer and dilution of liquid, the culture and screening of convenient microbe；
Nephelometer provided by the invention, is mounted on the outside of the area Ban Wei in the liquor removing workstation lower left corner, and Y-axis pallet is located at turbidity
Between the upper lower wall bar for counting U-shaped arm, microwell plate is placed on Y-axis pallet, the receiver and hair by the movement of U-shaped arm, above and below U-shaped arm
Emitter reads information, calculates the turbidity of liquid in microwell plate micropore, and then learns microorganism growth inside each hole of microwell plate
Situation, simply and easily learns the information of microorganism in micropore, and then facilitates researcher's screening；
The present invention is based on the screening techniques for the acquisition for single bacterial strain that automatical bacterium instrument provides, by using automatical
Bacterium instrument is screened, and comes out single strain isolation from complex sample, the single bacterial strain obviously grown is obtained, to grind
Research of the person of studying carefully to single bacterial strain.
Detailed description of the invention
Fig. 1 is the structural diagram of the present invention；
Fig. 2 is the schematic diagram after safety screen before present invention removal upper housing；
Fig. 3 is nephelometer schematic diagram of the invention；
Fig. 4 is that nephelometer of the present invention removes the schematic diagram after hood；
Fig. 5 is that nephelometer of the present invention removes the top view after hood.
In figure: 1, liquor removing workstation；11, liquid suction module；111, pipetting head；112, sucker rod；12, Y-axis pallet；
13, upper housing；14, X-axis driver；15, support board；16, Y-axis driver；17, the area Ban Wei；18, reject chute；2, nephelometer；
201, stepper motor；202, U-shaped arm；203, transmitter；204, receiver；205, left plate；206, right side plate；207, bottom plate；
208, preceding connecting plate；209, rear connecting plate；210, hood；211, screw rod；212, gag lever post；213, transmission gear；214, just
Beginning level sensor；215, control circuit board；216, USB interface.
Specific embodiment
To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention is made into one with reference to the accompanying drawing
Step illustrates.In the description of the present invention, it is to be understood that, term " on ", "lower", "front", "rear", "left", "right", "top",
The orientation or positional relationship of the instructions such as "bottom", "inner", "outside" is to be based on the orientation or positional relationship shown in the drawings, merely to just
In description the present invention and simplify description, rather than the device or element of indication or suggestion meaning must have a particular orientation, with
Specific orientation construction and operation, therefore be not considered as limiting the invention.In addition, what X-direction of the invention indicated
It is support board length direction, what Y direction indicated is the width direction of support board, and Z-direction instruction is perpendicular to support board
Direction, i.e. vertical direction.
Embodiment 1
A kind of automatical bacterium instrument, comprising:
Liquor removing workstation 1, as shown in Figs. 1-2, liquor removing workstation 1 include liquid suction module 11 and Y-axis pallet 12, liquid
Suction module 11 is mounted in the upper housing 13 of liquor removing workstation 1, and liquid suction module 11 is driven by X-axis driver 14 and realizes X
Axis is mobile, and Y-axis pallet 12 is mounted on the support board 15 of 1 bottom end of liquor removing workstation, and Y-axis pallet 12 is driven by Y-axis driver 16
Realize that Y-axis is mobile, Y-axis pallet 12 is equipped with 9 areas Ge Banwei 17, and the area Ban Wei 17 positioned at the lower left corner is hollow structure, the lower left corner
The area Ban Wei 17 in microwell plate is installed, have 384 micropores on the present embodiment microwell plate.Liquid suction module 11 includes multiple shiftings
Liquid head 111, pipetting head 111 are driven the suction for realizing liquid by sucker rod 112；Support board 15 is equipped with reject chute 18.
Nephelometer 2, as shown in Fig. 1,3,4 and 5, nephelometer 2 is mounted on 17 outside of the area Ban Wei in the lower left corner, and nephelometer 2 wraps
It includes and the U-shaped arm 202 for realizing that Y-axis is mobile is driven by stepper motor 201, Y-axis pallet 12 is located between the upper lower wall bar of U-shaped arm 202, U
The receiver 204 for being respectively equipped with transmitter 203 on the upper lower wall bar of arm 202 and cooperating with transmitter 203；Transmitter 203 is adopted
With the 650nm semiconductor laser with collimation lens, receiver 204 uses CdS photo resistance.
Nephelometer 2 further includes left plate 205, right side plate 206, bottom plate 207, the preceding connecting plate 208 for constituting nephelometer skeleton
With rear connecting plate 209, stepper motor 201 and U-shaped arm 202 are located in skeleton, and hood 210 is equipped on the upside of skeleton.Left plate 205
Screw rod 211 and gag lever post 212 are equipped between right side plate 206, stepper motor 201 passes through 213 connecting screw 211 of transmission gear, spiral shell
The threaded hole that bar 211 passes through U-shaped 202 downside of arm is threadedly coupled with U-shaped arm 202, and gag lever post 212 passes through the logical of U-shaped 202 downside of arm
Hole.It is equipped with initial level sensor 214 close to 205 end of left plate on connecting plate 209 afterwards, 207 are equipped with control circuit board on bottom plate
215, right side plate 206 is equipped with the USB interface 216 of connection control circuit board 215.
Control system, control system include controller, and controller is connect with liquor removing workstation and turbidimetric signal, control
Device is conventional single-chip microcontroller, is implanted into process control liquor removing workstation and turbidimetric operation, and the present embodiment is selected
AtmelATmega328p formula single-chip microcontroller.Specifically, single-chip microcontroller controls the liquid of liquid suction module 11 for liquor removing workstation 1
Suction, single-chip microcontroller control X-axis driver 14 and drive liquid suction module 11 mobile in X-direction, and single-chip microcontroller controls Y-axis pallet 12
It is mobile in Y direction；For nephelometer 2, single-chip microcontroller control transmitter 203 emits light, according to the reception information of receiver 204
The turbidity of liquid in micropore is calculated, single-chip microcontroller control stepper motor 201 drives U-shaped arm 202 in the mobile reading micropore of Y direction
Middle fluid information.
Embodiment 2
The present embodiment on the basis of embodiment 1, provides a kind of based on the single bacterial strain of 1 automatical bacterium instrument of embodiment screening
Screening technique, the primary sample of the present embodiment is the complex sample containing Intestinal Anaerobic Bacteria, and the purpose of the present embodiment is to obtain
The single bacterial strain that having in primary sample obviously grows, the method for the present embodiment including the following steps:
Step S1 prepares primary sample, is diluted primary sample with the liquor removing workstation of automatical bacterium instrument, after dilution
YCFA culture medium is added in primary sample, obtains media samples, is divided media samples with the liquor removing workstation of automatical bacterium instrument
It is attached in the micropore of 384 microwell plates；
Step S2, by the media samples in microwell plate, mesh after being cultivated 5 days in the constant incubator outside automatical bacterium instrument
Strain growth is marked, the cultivation temperature in constant incubator is 37 DEG C；
Step S3, the media samples that culture is completed are read each on microwell plate with the nephelometer on automatical bacterium instrument
The turbidity value of culture medium in micropore, and generate picking file；
Step S4, automatical bacterium instrument set liquor removing workstation and choose bacterium minimum according to nephelometer testing result in picking file
OD value, liquor removing workstation carry out picking to the culture medium in the micropore for being greater than minimum OD value, obtain single bacterial strain.
The results are shown in Table 1 after the condition and screening of screening:
The screening of 1 Intestinal Anaerobic Bacteria strain of table
By using bacterium instrument screening microorganism is chosen, the separation flux of microbial strains can reach 100-400/hour, answer
The 16S rRNA gene of bacterial strain after picking in above-mentioned table is sequenced with MALDI-TOF and/or PCR, find pure bacterium ratio 80% with
On.
In above-mentioned steps S1 and S4, automatical bacterium instrument, which is diluted with picking, to be aspirated using the liquid of liquor removing workstation 1
What the pipetting head 111 of module 11 was realized, pipetting head 111 is provided with multiple, and microwell plate is placed on Y-axis pallet 12, on microwell plate
Multiple micropores are evenly arranged with, drives the microwell plate on Y-axis pallet mobile in Y direction by Y-axis driver 16, is driven by X-axis
Device 14 drives the pipetting head 111 of liquid suction module 11 mobile in X-direction, by Z axis driver drives pipetting head 111 vertical
Direction is mobile, and by three-shaft linkage, pipetting head 111 is diluted to the liquid in micropore on microwell plate and picking.
In above-mentioned steps S3, the turbidity value that nephelometer reads culture medium in each micropore of microwell plate includes following step
It is rapid:
Microwell plate, is placed in the lower left corner position of Y-axis pallet 12 by step S31, and the U-shaped arm 202 of nephelometer 2 initializes first
It returns at initial level sensor 214, not between transmitter and receiver, transmitter 203 does not work microwell plate, and reading connects at this time
The signal for receiving device 204 is simultaneously transmitted to control system, and control system is by signal framing transmission zero point at this time；Then transmitter 203
Work, receiver 204 receive signal and are transmitted to control system, and signal framing transmission at this time is worked to the last minute；Transmission work to the last minute and
Zero point is used to calculate the transmitance of liquid.
Step S32, the microwell plate on Y-axis pallet 12 is mobile with 2 direction of trend nephelometer by Y-axis driver 16, works as micropore
First round of plate stops movement after being in right above the transmitter 203 of U-shaped 202 lower part of arm, and then U-shaped arm 202 is by stepping electricity
Machine 201 drives according to the program of setting from initial position to distally gradually moving, i.e., mobile to X-direction, when U-shaped arm 202 is every
After reaching below a hole, which is transmitted to control by the light signal issued using the transmitter 203 that receiver 204 receives
The signal and zero point and working to the last minute is made comparisons, is calculated as the transmissivity in the hole, is then convert into optical density by device processed, controller, this
In embodiment, controller is single-chip microcontroller.
Step S31 repeats step S31-S32, completes the reading to entire microwell plate, and generate picking text by controller
Part.
The basic principles, main features and advantages of the present invention have been shown and described above.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its
Equivalent thereof.
Claims (9)
1. a kind of automatical bacterium instrument characterized by comprising
Liquor removing workstation (1), the liquor removing workstation (1) include liquid suction module (11) and Y-axis pallet (12), the liquid
Suction module (11) is mounted in the upper housing (13) of liquor removing workstation (1), and liquid suction module (11) is by X-axis driver (14)
It drives, the Y-axis pallet (12) is mounted on the support board (15) of liquor removing workstation (1) bottom end, and Y-axis pallet (12) is driven by Y-axis
Dynamic device (16) drive, and the Y-axis pallet (12) is equipped with multiple areas Ban Wei (17), and the area Ban Wei (17) positioned at the lower left corner is hollow
Structure；
Nephelometer (2), the nephelometer (2) are mounted on the outside of the area Ban Wei (17) in the lower left corner, and the nephelometer (2) includes by step
The U-shaped arm (202) driven into motor (201), Y-axis pallet (12) is located between the upper lower wall bar of the U-shaped arm (202), described U-shaped
Transmitter (203) and the receiver (204) with transmitter (203) cooperation are respectively equipped on the upper lower wall bar of arm (202)；
Control system, the control system include controller, the letter of the controller and liquor removing workstation (1) and nephelometer (2)
Number connection.
2. a kind of automatical bacterium instrument according to claim 1, which is characterized in that the nephelometer (2) further includes that composition is turbid
Left plate (205), right side plate (206), bottom plate (207), preceding connecting plate (208) and the rear connecting plate (209) of degree meter skeleton, it is described
Stepper motor (201) and U-shaped arm (202) are located in skeleton, are equipped with hood (210) on the upside of the skeleton.
3. a kind of automatical bacterium instrument according to claim 2, which is characterized in that the left plate (205) and right side plate
(206) screw rod (211) and gag lever post (212) are equipped between, the stepper motor (201) passes through transmission gear (213) connecting screw
(211), the threaded hole that the screw rod (211) passes through on the downside of U-shaped arm (202) is threadedly coupled with U-shaped arm (202), the gag lever post
(212) through-hole on the downside of U-shaped arm (202) is passed through.
4. a kind of automatical bacterium instrument according to claim 3, which is characterized in that close to left on connecting plate (209) after described
Side plate (205) end is equipped with initial level sensor (214), and (207) are equipped with control circuit board (215) on the bottom plate, right side plate
(206) it is equipped with the USB interface (216) of connection control circuit board (215).
5. a kind of automatical bacterium instrument according to claim 4, which is characterized in that the transmitter (203) uses 600-
605nm LED and lens group carry out transmitting light collimation or the 650nm semiconductor laser with collimation lens, the receiver
(204) CdS photo resistance is used.
6. a kind of automatical bacterium instrument according to claim 1, which is characterized in that the liquid suction module (11) includes more
A pipetting head (111), the pipetting head (111) are driven the suction for realizing liquid by sucker rod (112).
7. a kind of automatical bacterium instrument according to claim 1, which is characterized in that the support board (15) is equipped with reject chute
(18)。
8. a kind of microbe to screen method of automatical bacterium instrument, which is characterized in that this method is used for the acquisition of single bacterial strain, including
Following steps:
Step S1 prepares primary sample, is diluted primary sample with the liquor removing workstation of automatical bacterium instrument, will be original after dilution
Culture medium is added in sample, obtains media samples, media samples are dispensed into micropore with the liquor removing workstation of automatical bacterium instrument
In the micropore of plate；
Step S2, by the media samples in step S1 in microwell plate, constant temperature incubation is to mesh at a temperature of aimed strain growth
Mark strain growth；
Step S3 will cultivate the media samples of completion in step S2, with the nephelometer on automatical bacterium instrument, read on microwell plate
The turbidity value of culture medium in each micropore, and generate picking file；
Step S4, automatical bacterium instrument set liquor removing workstation and choose bacterium minimum OD value according to nephelometer testing result in picking file,
Liquor removing workstation carries out picking to the culture medium in the micropore for being greater than minimum OD value, obtains single bacterial strain.
9. the microbe to screen method of automatical bacterium instrument according to claim 8, which is characterized in that in above-mentioned steps S3,
Nephelometer read the turbidity value of culture medium in each micropore on microwell plate the following steps are included:
Microwell plate, is placed in the lower left corner position of Y-axis pallet (12) by step S31, and turbidimetric U-shaped arm (202) initialization returns to
At initial level sensor (214), and read the signal of no light and illumination without plate when, it is determined as the zero point of transmission and work to the last minute, use
In calculating transmissivity；
Step S32, Y-axis pallet (12) is mobile to nephelometer direction, when the first round for being moved to microwell plate is in U-shaped arm
(202) stop movement behind the light source surface of lower part, then U-shaped arm (202) is according to the program of setting from initial position to distally
It gradually moves, controller reads the transmissivity in the hole behind the lower section of one hole of every arrival, and is converted into OD value；
Step S33 repeats step S31-S32, and nephelometer completes the reading to entire microwell plate, and generates picking text by controller
Part.
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Application publication date: 20190212 Assignee: Sichuan anaerobic Biotechnology Co.,Ltd. Assignor: BIOGAS SCIENCE Research Institute MINISTRY OF AGRICULTURE|Cheng Lei Contract record no.: X2021510000008 Denomination of invention: An automatic bacteria picking instrument and microbial screening method Granted publication date: 20200908 License type: Exclusive License Record date: 20210322 |