CN109554292A - Manifold Light Way device and microorganism high-pass culture and detection device - Google Patents
Manifold Light Way device and microorganism high-pass culture and detection device Download PDFInfo
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- CN109554292A CN109554292A CN201910082576.XA CN201910082576A CN109554292A CN 109554292 A CN109554292 A CN 109554292A CN 201910082576 A CN201910082576 A CN 201910082576A CN 109554292 A CN109554292 A CN 109554292A
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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
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/16—Vibrating; Shaking; Tilting
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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
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
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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
- C12M41/32—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of substances in solution
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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
- C12M41/48—Automatic or computerized control
Abstract
The present invention relates to laboratory microbe research fields, a kind of Manifold Light Way device and microorganism high-pass culture and detection device are disclosed, Manifold Light Way device, improved shaking device including high-throughput detection microbiological specimens concentration and suitable for Manifold Light Way device or the composite microporous plate of automation equipment.The detection device using optical splitter and forward scattering optical path in conjunction with mode come the concentration for detecting sample in each hole on microwell plate of staggering the time, avoid common multi-channel detection probe simultaneously to multiple sample detections when adjacent holes signal the shortcomings that interfering with each other.The work of the optical path is made of the coordinated control of optical splitter, infrared light emission circuit and mid-infrared optical receiving circuit, infrared light emission circuit uses dedicated constant-current source chip controls, both can guarantee infrared light-emitting diode using safe and service life, while also can guarantee the infrared light power stabilising of transmitting.
Description
Technical field
This technology belongs to laboratory microbe research field, be related to it is a kind of can Manifold Light Way device and microorganism high-pass
Culture and detection device, Manifold Light Way device, improved shaking device including high-throughput detection microbiological specimens concentration with
And suitable for Manifold Light Way device or the composite microporous plate of automation equipment.
Background
In biological technology products other than the maximum bio-pharmaceuticals of accounting, on industrial market, fine chemicals and biology
Fuel alsies occupy consequence.More and more industrial applications and product are directed to or derived from biocatalytic reactions.These
Most of emerging product is generated in microorganism or cell culture fermentation.In addition, that emerges in large numbers in genetic engineering is various new
Research tool is used to manipulation cell to generate new biomolecule.Directed evolution, systems biology and synthetic biology field
Road has been paved for bioprocess novel in design and with more high efficiency from bottom to top.It is designed in microorganism or cell
New metabolic pathway enables more competitive bioprocess to replace or improve chemical production processes.Chemistry and pharmaceutical industry by
To the pressure of market competition, each company is all developing new product with speed as fast as possible, to shorten time to market (TTM), and
And the term of a patent of new pharmacy also limits and these companies is forced to put out a new product in market early stage to obtain bigger market reward.
Very fast and efficiently tool, new research tool also become promotion biology to the speed needs of these bioprocess of Speeding up development
The necessary condition of process research and development.
The these requirements of biotechnology have expedited the emergence of the appearance of high flux screening (HTS) technology.Invention is in the micropore of nineteen fifty-one
Plate (or microtiter plate, MTPs) most starts to be mainly used for meeting pharmacy, chemistry and demand high-throughput in basic research.Pass through
More sample wells are concentrated on same area, the flux of microwell plate is continuously increased, and is developed from 6,12,24,48,96,384,1536
For 3456 holes.Present microwell plate has become one of the standard tool of drug discovery high throughput experiment.
Easy to operate since it is low with use cost, flux is high, the feature of energy and most of automation equipments compatibility,
In past more than ten years, application also more and more common and triangular flask of the various microwell plates (hole 24-384) in culture microorganism
Equally, nowadays microwell plate has become most common bioreactor in scientific research and industrial bio process development.
It is its limited maximum oxygen transfer ability (OTR that the distinct disadvantage of microculture is carried out in microwell platemax).Therefore,
Microwell plate is limited primarily to establish the clone library of Escherichia coli and yeast for microculture and preservation initial stage.Due to large intestine bar
Bacterium and most of yeast can be in anaerobism or grown under hypoxic conditions, so oxygen supply deficiency is usually to culture effect without too
It is big to influence.However, in aerobic bacteria incubation, the case where just will appear undergrowth in microwell plate.If high flux screening
(HTS) or bacterial strain development process purpose is screening the production bacterial strain for being used for aerobic fermentation, then the microorganism for being necessary for screening provides
Enough dissolved oxygens.Otherwise, bacterium will fail, even if screening successfully, the later period expands in the fermentor of oxygen supply abundance
It may also can be invalid when culture is to obtain more products.
In recent years, with the further investigation that the bioengineering parameter characterization aspect to dynamic microwell plate carries out, discovery influences
The determinant of gas-liquid oxygen transmission speed is whether the sufficiently large liquid film of area can be formed on the inner wall of hole in micropore.With micro-
The reduction of hole cross-sectional area, surface tension can apply increasing resistance to the flowing of fluid in microwell plate, in addition capillary
The superposition of Guan Li more hinders the free diffusing and forced flow of molecule.To which the liquid film of larger area cannot be formed, and then lead
Cause the gas-liquid oxygen transmission speed under bigger amplitude and common shaking frequency very low.It is passed so to reach same gas-liquid oxygen
Effect is passed, under conditions of identical shaking radius, for 96,384 and 1536 orifice plates, compared to traditional triangular flask or culture bottle
Then need higher shake speed.
For inventor's discovery under the conditions of the shaking of high speed circumference, liquid forms film on the arc-shaped wall of 48 hole microwell plates
When, mass tranfer coefficient (kL) 0.8m/h can be up to, hence it is evident that it is better than 96 orifice plates or triangular flask.Therefore, 48 orifice plates can be used as microorganism
The powerful of culture and expression study has both 96 orifice plate high throughputs and the tradition shaking higher gas-liquid oxygen transmission speed of triangular flask
The advantages of.
Realize that the circumference of culture vessel shakes in the prior art, the most commonly used is horizontal circumference shaking table cultures in the lab
Case, this has (1) three point of shake system, is made of three bearings, three bearings shaking platform below triangle in, wherein
One axis is driven by motor, or (2) central actuator, supports shaking platform in conjunction with quadrangle.In both cases, internal
Counterweight is directly placed at below shaker platform so that during shaking shaft reached up in horizontal plane side it is flat completely or partially
Weighing apparatus, avoids cabinet from acutely shaking.Since the driving device of traditional most shaking table culture casees or microwell plate rocker is all located at tightly
Suffer shaking platform following location directly, the space not moved back and forth to mobile probe, therefore this drive system or counterweight
In the presence of, shaking platform below there is no too big space that can be utilized.And only one axis is driven by motor mostly, and it is other
Axis follows shaking platform passive rotation.When being run using common shaking frequency, mechanical part is easy by extra pressure
Abrasion, especially at higher rotational speeds, this abrasion can shorten the service life of shaking device.Microorganism high-pass culture
When, if necessary to their growing state of real-time monitoring or special condition of culture (uniform illumination is such as provided) is provided, is needed
Device and/or lighting device are placed below platform, common shaking table does not have enough spaces then.Therefore, existing circumference
Shaking device can only adapt to the demand of microorganism high-pass culture, not can be implemented simultaneously the need of the concentration of real-time measurement microorganism
It asks.
In addition, real-time measurement microorganism concentration in the prior art, common research method be using microplate reader or
The micropore board detector of similar microplate reader principle.This method is by measuring transmitted light and forward scattering luminous intensity in common microwell plate
To detect optical density, optical density and sample concentration proportional variation in a certain range.But due to the shaking of common microplate reader
Mode is substantially the linear shaking of low speed, and shaking intensity is mild, frequency is low, can only achieve the purpose that sample is simply mixed, and
It cannot make to reach higher gas-liquid oxygen transmission speed in hole.And common microwell plate is made of upper cover and orifice plate, uses microwell plate
When cultivating microorganism, if covering tightly upper cover, microorganism in hole cannot be made to obtain sufficient oxygen, will affect growth of aerobic microorganisms
Effect；If not covering tightly upper cover, the liquid in hole can evaporate, and the liquid evaporation speed and oxygen of apertura lateralis and inboard aperture are handed over
Throw-over degree is different, and leading to cultivation results, there are apparent edge effects.Although can be with using hydrophobic permeable membrane covering microwell plate
Liquid evaporation is prevented, while guaranteeing gas exchanges, but existing hydrophobic permeable membrane light transmittance is poor, optical path is will affect and passes through to test sample
Product, to influence detection effect.
Chinese patent application 201080009279, before the denomination of invention equipment of biological sample " analysis " discloses a kind of use
To the technology with bacteria suspension concentration in Scattering Measurement measurement microwell plate, invention bacteria suspension concentration suitable for microwell plate
Observational measurement so that realization quickly and in high-throughput test sample whether there is bacterium, and completes parting or antibacterial inspection.Due to
The invention haves the shortcomings that similar with microplate reader, therefore the invention is only used for the qualitative detection of culture, i.e., examines for medicine
Disconnected purposes, is not appropriate for the high quality culture of aerobic microbiological.
The Chinese invention patent application 201310246480, " shaking table device of contactless multichannel on-line checking fermentation liquid
And test method " device that multichannel real-time detection culture bacterium solution is realized using spectrophotometer principle is disclosed, the device is logical
It crosses detection and transmits the optical density of special triangular flask fluid column to realize the growing state of multiple culture vessel real-time detection microorganisms.So
And this method needs that laser emitter and receiver is arranged in each culture vessel two sides, and culture vessel differs markedly from
The culture environment of microwell plate cannot achieve the purpose of high-throughput microwell plate culture and real-time detection.
" a kind of non-intrusion type biomass for shaking flask culture exists for Chinese patent application 201510551900, denomination of invention
Line detector ", the device realize the on-line checking of shaking flask culture bacterial concentration using back scattering optical measurement techniques, can be not
In the case where interrupting culture device, the concentration of bacteria suspension in shaking flask is detected.But when sample concentration concentration is smaller or training
When feeding container is smaller, vulnerable to the interference of chamber wall reflected light, it is only suitable for the biggish triangular flask of volume, is not suitable for microwell plate
Detection, and if each container need install additional a detection device, this for microwell plate, that is, increase equipment at
Originally, increase systematic error, also have great difficulty on the implementation.
" one kind is used for microorganism high-pass culture and its big concentration for Chinese patent application 201610341529, denomination of invention
The self-reacting device of range real-time detection ", the device constitute the compound of a dual detector using transmission and back scattering optical path
Optical path achievees the purpose that single detection probe detects multiple samples in conjunction with turntable rotation.But the device is difficult to will test flux
100 or more are expanded to, and it is single vial that applicable culture vessel is set in repacking, is more than 100 for detection flux
A experiment, operating, it is convenient to can not show a candle to microwell plate.
" one kind is high for microorganism for the present inventor's prior authorization Chinese invention patent 201610341529.9, denomination of invention
The self-reacting device of flux culture and its big concentration range real-time detection " discloses a kind of by transmitted light path and backward astigmatism road knot
It is combined, the two shares a light source (light emitting diode) to complete the detection probe of real-time detection microculture concentration.
In addition, the invention also discloses the microorganism high-pass culture apparatus comprising the detection probe, include high-throughput liquid agitation dress
It sets, sample bottle tray transmission, temperature control equipment etc..The invention is able to achieve the high-throughput culture and real-time detection of microorganism
The purpose of concentration, but the detection probe of the invention is mainly the detection advantage of comprehensive transmitted light and rear orientation light, is passed through simultaneously
It improves the agitating device (containing transmission device) of fluid nutrient medium and temperature controls and the combination of Concentration Testing probe, not
The solution for how realizing the dissolved oxygen of circumference shaking and guarantee culture medium abundance can be provided.In addition, the invention is mainly for appearance
Measure be not appropriate in 10-50 milliliter of culture bottle capacity in microwell plate of several hectolambdas to several milliliters liquefied gas exchange with
Prevent the needs of evaporation.
To sum up, in the prior art, when carrying out microculture, since most of microbe needs to disappear in the metabolic process
Oxygen consumption gas, the gaseous products such as discharge carbon dioxide, the gas exchanges in culture solution can be accelerated by ceaselessly shaking culture solution, be made micro-
Biology obtains more oxygen, is uniformly distributed nutriment, microorganism can also be made always to be in suspended state, to allow at microorganism
In than convenient growing environment.Conventional method is that the culture vessel of the larger capacities such as test tube, triangular flask is placed on to shaking training
It supports in case, but since culture vessel is constantly in shaking state, it is not easy to it pops one's head in and integrates with Concentration Testing, it is more difficult to reach
The high-throughput of a samples up to a hundred is detected simultaneously to require.And triangular flask culture aerobic microbiological is used in using shaking incubator
When, even if the additive amount of culture medium only has 1/5th, the maximum oxygen transfer ability of culture medium is also limited, the dissolved oxygen in culture medium
It is not able to satisfy the growth of part aerobic microbiological.If replacing the culture vessel progress of larger capacity high-throughput micro- using microwell plate
Biological culture, then existing circumference shakes device or can not achieve efficient circumference shaking, while internal and bottom lacks appearance
Receive the space of detection device and lighting device.Meanwhile existing detection device, it is also necessary to consider the gas of sample liquids in microwell plate
Liquid evaporation is exchanged and prevented, while also to guarantee that light path is not blocked or interfered in microwell plate.Meanwhile for high pass
Amount, the culture minimized, it should make every effort to single or single-row Concentration Testing probe and a large amount of samples are detected, reduce detection to the greatest extent
Probe quantity, and realize the automation of entire detection process.
It can be seen that either using the method or existing real-time online detection side of microplate reader or similar microplate reader
Method is all unable to satisfy the purpose that high-throughput culture and real-time online measurement of concetration are carried out using microwell plate.
Summary of the invention:
Therefore, the problem to be solved by the present invention is that: it is dense for high-flux microorganism culture and real-time detection microorganism
The instrument of degree, it is therefore desirable to realize that single or single-row detection probe detects multiple or multiple row sample, realization entirely detected
The automation and high throughput of journey, and increase the maximum oxygen transfer ability value of culture medium as far as possible, so that the prior art be avoided to exist
Defect.
Therefore, the purpose of the present invention is to the shortcoming of existing equipment to be overcome, provide a kind of full-automatic, high accuracy,
The self-reacting device of microorganism high-pass culture and its concentration real-time detection that is high-throughput, intuitively changing.
In order to achieve the above technical purposes, the present invention adds on the basis of traditional transmitted light or forward scattering light detection method
Enter the high speed shaking device and multichannel Concentration Testing probe of microwell plate, it is ventilative high saturating using the height for being more suitable for microculture
The deep-well plates of light.
Therefore, one of principle of the invention is, the present invention increases optical path choosing on the basis of traditional microplate reader light path
Device is selected, the detection so that each optical path is independently staggered the time, the signal interference of adjacent optical path when avoiding because while detecting.Meanwhile increasing inspection
End and receiving end distance are surveyed to adapt to the sample of microwell plate or bigger light path with higher size.
The two of the principle of the invention are, in the technology of traditional eccentric shaking, have invented that the degree of eccentricity is smaller, and shake speed is more
High technology realizes high-throughput culture of each sample under the conditions of uniform shaking in conjunction with detection probe horizontal linear track
Automatic detection with single detection probe to multiple row sample.
The three of the principle of the invention are, in order to guarantee in microwell plate higher oxygen transfer ability between liquid and gas, more
Be conducive to the growth of aerobic microbiological and the expression of product, avoid edge effect caused by common lid for micro plate, has invented suitable
Close the ventilative light transmission upper cover of microwell plate.The upper cover is made of metal cover, hydrophobic permeable membrane, light-transmitting plate and silicagel pad.In conjunction with partly leading
The technologies such as body temperature control and the ventilative light transmission upper cover of microwell plate and data acquisition, analysis, output software, realize microorganism high-pass
Culture, the full-automation of concentration real-time detection and data analysis.
Therefore, the first purpose of the invention is to provide a kind of multi-pass that can detect high-flux microorganism sample concentration simultaneously
Road light path device, the device include: light emitting diode 1 and the optical fiber 2 that communicates therewith, the optical splitter 3 be connected to optical fiber 2, and are divided
Multichannel optical fiber 4 and the Concentration Testing probe 23 that light device 3 is connected to, wherein
Concentration Testing probe 23 includes the fiber fixed seat 5 as upper arm and the photodiode fixing seat 8 as lower arm,
Fiber fixed seat 5 is connected to above-mentioned multichannel optical fiber 4, and photodiode fixing seat 8 is equipped with corresponding with the optical path of multichannel optical fiber 4
Multichannel photodiode 9, and upper and lower arms respectively correspond the vertical upper and lower that microwell plate is arranged in, and lead to each other
It crosses and is vertically connect with the connector of upper lower arm, to form the Concentration Testing probe of the U-shaped opening of horizontal direction；
Light emitting diode 1 issues the optical coupling entering light fibre 2 of fixed wave length, and light can be assigned to finger after optical splitter 3
Fixed multichannel (such as 8-16 root optical fiber) optical fiber 4, vertically penetrates the more of microwell plate 6 from a plurality of optical path of 4 channel emission of optical fiber
The sample in fluid nutrient medium 7 in a micropore, and scattered through the sum of sample by sample by 23 detection of Concentration Testing probe
Forward scattering light, and optical signal is converted to electric signal, then convert electrical signals into digital signal through circuit, whole process is
Concentration Testing optical path is formed, to complete the detection of high-flux microorganism sample concentration.
In any of the above-described embodiment, the upper lower arm of the detection probe 23 fixes the optical fiber 4 in 8-16 channel respectively
With photodiode 9, the optical fiber 4 and photodiode 9 in preferably 12 channels.
In any of the above-described embodiment, the upper lower arm spacing distance of the detection probe 23 is 20-50mm, wherein it is preferred that
It is 40mm.
In any of the above-described embodiment, the light emitting diode 1 is hair of the peak wavelength range in 600-1400nm
Optical diode, preferably 850nm.
Second purpose of the invention is to provide the automation equipment of a kind of microorganism high-pass culture and concentration real-time detection,
Including superposed aforementioned Manifold Light Way device, the shaking platform of the multiple microwell plates of carrying at middle part, the microwell plate of lower part
Device is shaken, and is located at the detection probe mobile device of bottom, wherein
(1) shaking platform 18 is located between the U-shaped opening of Concentration Testing probe, and Concentration Testing probe can be on shaking platform
Multiple microwell plate upper horizontals it is mobile；
(2) microwell plate being fixedly connected with shaking platform 18 shakes device, and the motor 28 including bottom is located on motor 28
The bearing (setting) plate 12 of side, three eccentric shafts 17 of asymmetric setting on bearing (setting) plate 12, each 17 middle section of eccentric shaft passes through same
Step band 11 is connected with each other, and 17 upper section of eccentric shaft is fixedly connected with shaking platform 18, and synchronous belt 11 is fastened the motor on plate turn
On axis 14, when the rotation of 28 driving motor shaft 14 of motor, shaft 14 drives synchronous belt to drive multiple 17 rotations of eccentric shaft, thus
Top shaking 18 high speed circumference of platform shaking is driven, it is final to drive the shaking of 6 high speed circumference of microwell plate；
(3) detection probe mobile device includes the linear slide rail 36 that stepper motor 39 controls, the sliding rail and shaking platform 18
It is distributed in vertical equity, one end is connected with stepper motor 39, and the other end passes through the Concentration Testing of fixing seat fixing seat 30 and top
Upper arm or the lower wall connection of probe 23 can drive the Concentration Testing probe 23 of top when stepper motor 39 drives sliding rail mobile
U-shaped opening from shaking platform microwell plate vertically side moving.
In one embodiment, in the microwell plate shaking device, eccentric shaft 17 is equipped with synchronizing wheel 11, and by same
Step band 11 is connected with each other；Motor 28 is fixed on motor fixing plate 15 by flange, and motor fixing plate 15 is fixed on bearing and fixes
Plate 12, motor fixing plate 15 is equipped with machine shaft 14, and is fixedly connected with synchronizing wheel 13, so that motor 28 passes through synchronizing wheel
13, synchronous belt 11 is connected with the synchronizing wheel 11 of three eccentric shafts 17.
In a preferred embodiment, the fixed lower bearing 22 in three 17 lower ends of eccentric shaft, lower bearing 22 are fixed on bearing
12 in fixed plate, the fixed upper bearing (metal) 21 in 17 upper end of eccentric shaft, upper bearing (metal) 21 is fixed on shaking platform 18, wherein above and below eccentric shaft
The axis of the circular motion at both ends is in two parallel lines of spacing 1-3mm, and synchronous pulley 13 and counterweight 16 are fixed on eccentric shaft 17
Lower end, 16 center of gravity direction of counterweight and 17 offset direction of eccentric shaft are exactly the opposite in the horizontal direction, with offset or partial offset because
Shaking platform bias shakes the uneven centrifugal force to be formed；When motor 28 drives the synchronizing wheel 13 being fixed on machine shaft 14 to revolve
Turn, and then drive synchronous belt 11, and then drives the synchronizing wheel 10 being fixed on eccentric shaft 17 to rotate, and then eccentric shaft 17 is driven to revolve
Turn to rotate with the counterweight 16 on eccentric shaft 17.Since the upper and lower ends of eccentric shaft 17 have, 1-3mm's is axially eccentric, so eccentric shaft
17 upper ends can do circle rotation by radius of 1-3mm, and then shaking platform 18 is driven to do circumference shaking by radius of 1-3mm, most
Realize that microwell plate 6 does circumference shaking by radius of 1-3mm eventually.
In another embodiment, in the mobile device, positioning caliper 37 with groove is installed in 36 side of sliding rail, and sliding rail is sliding
Fixed concentration detection probe 23 and the optoelectronic switch 38 for converting optical signals to electric signal, positioning caliper 37 and photoelectricity on platform
The stop place when cooperation of switch 38 is for positioning Concentration Testing 23 detection of probe.
In any of the above-described embodiment, shaking platform 18 can place 2-8 microwell plate, in vertical on shaking platform 18
To 2 × 1,2 × 2,2 × 3,2 × 4 arrangements, preferably 2 × 2 arrangements, used microwell plate 6 is 24 holes (hole is in 4 × 6 arrangements), 48
Hole (hole is in 6 × 8 arrangements) and 96 holes (hole is in 8 × 12 arrangements) microwell plate 6, preferably 48 hole microwell plates 6.In any of the above-described tool
In body embodiment, 6 height of microwell plate is 20-50mm, preferably 40mm.
In any of the above-described embodiment, 28 revolving speed of motor is 1000-3000 revs/min, preferably 1500-2000
Rev/min, the balance of maximum oxygen transfer ability in the stability and micropore to reach shaking device.
In any of the above-described embodiment, the number of recesses of positioning caliper 37 is 6-24, preferably 16, each recessed
Groove edge and the alignment of the 6 each column hole center of circle of microwell plate.
Third purpose of the present invention is to provide a kind of suitable for the composite microporous of Manifold Light Way device or automation equipment
Plate, including the hot lid 25 of microwell plate, microwell plate, microwell plate ventilating cover, wherein
Hot lid 25 can cover one or more microwell plates, and the heat passed through convenient for light path is equipped in micropore corresponding position
Cap bore 42, hot lid 25 can heat microwell plate ventilating cover；
Microwell plate ventilating cover successively includes casting coping 44, hydrophobic permeable membrane 45, light-transmitting plate 46 and silicagel pad from top to bottom
47, wherein casting coping 44 be used to support, fixed ventilation lid, hydrophobic permeable membrane 45 allow micropore inside and outside gas exchanges, light transmission
Plate 46 penetrates the fluid sample in micropore by hot cap bore 42 convenient for light path, and silicagel pad 47 is each for sealing microwell plate
Independent hole.
In one embodiment, it is equipped with heating sheet 41 on the inside of hot lid 25, and can be close to casting coping 44, it can be to metal
Top cover 44 is heated, it is ensured that the temperature of light-transmitting plate 46 is 1-2 degrees Celsius higher than culturing room's inner cavity temperature, is conducive to so for a long time
When cultivating microorganism, the vapor excessive evaporation in microwell plate is avoided, while avoiding forming vapor on light-transmitting plate 46, influenced
Testing result.In a specific embodiment, 25 edge of hot lid is equipped with hot lid shaft 40 and hot cover lock button hole 43, wherein
Hot lid shaft is to rotate heat lid can, convenient for picking and placing microwell plate, after having picked and placed microwell plate, covers hot lid, and hot cover latch can be with
Heat lid is fixed, hot Gai Songdong during avoiding high speed from shaking.
In another embodiment, the micropore on microwell plate is completely covered in the light-transmitting plate 46, sets in micropore marginal end
Strip-shaped hole is set, which covers hydrophobic permeable membrane 45, so that the gas inside and outside micropore passes through hydrophobic permeable membrane 45 and strip-shaped hole
It swaps, while preventing vapor from evaporating；The casting coping 44 and silicagel pad 47 for covering microwell plate are in the smaller micropore of diameter
Region is hollow structure, and hole is equipped at the strip-shaped hole for corresponding to light-transmitting plate 46, to avoid closing or blocking on ventilated membrane
Gas exchanges access, so that light-transmitting plate 46 is hanging down relative in the region of the 3 millimeters of radiuses in micropore each on microwell plate center
Histogram is not blocked by casting coping 44, hydrophobic permeable membrane 45, silicagel pad 47 upwards, is unobstructedly passed through in micropore convenient for optical path
The heart, to complete the optical detection to the sample of fluid nutrient medium 7 in micropore.
In one embodiment, the light-transmitting plate 46 is the light transmission thin plate of high transparency, to reduce optical path across institute's band
The optical path loss come, preferably the organic syntheses thin plate such as the glass plate of high transparency, crystal slab, quartz plate, PC plate or PET sheet.
In one embodiment, the ventilated membrane 45 is hydrophobic air permeable film, to prevent water steam to be aggregated.It is excellent at one
It selects in embodiment, the aperture of the hydrophobic permeable membrane 45 is no more than 0.45 micron, can be certainly convenient for gases such as oxygen, carbon dioxide
By penetrating, and prevent vapor and microorganism from penetrating.
4th purpose of the invention is to provide a kind of automation for microorganism high-pass culture and concentration real-time detection
It further include conductor temperature control except set composite, including above-mentioned Manifold Light Way device, automation equipment, composite microporous plate
Device and computer processed control software, wherein
The conductor temperature control device includes Peltier semiconductor chilling plate 33, refrigeration head 32, cooling fin 34 and two
Group radiator fan 31 and 35, refrigeration head 32 are located in culturing room at cavity wall, front end connection fan 32, and semiconductor chilling plate 33
Between refrigeration head 32 and cooling fin 34, fan 35 is connected on the outside of cooling fin 34.
In any of the above-described embodiment, culture room temperature control can be existed in conductor temperature control device
10-60 degrees Celsius, it can satisfy the culture from thermophilic cold, room temperature and mesophilic microorganism.Temp probe 26 on 24 wall of inner cavity measures
It cultivates indoor temperature and feeds back to control circuit.
In any of the above-described embodiment, wherein microwell plate shaking device, Concentration Testing probe and conductor temperature control
Device passes through software control.On computers, computer is connected by RS232 connection with instrument host the software installation.On software
Following functions may be implemented:
1) target temperature intracavitary in culturing room is set；
2) motor speed is set；
3) setting Concentration Testing is popped one's head in detection interval time；
4) each sample concentration of independent real-time display corresponds to the change curve of time, and each sample has independent display window
Mouthful；
5) real-time display temperature corresponds to the change curve of time；
6) change curve for corresponding to the time to a plurality of concentration compares；
7) it is directed to various concentration unit, there is automatic calibration function；
8) detailed sample message can be marked to each sample；
9) for the sample size detected every time, corresponding curve display window can be selected by sample placement location；
10) sample can be grouped automatically according to sample message, and be averaged to the detected value of parallel sample；
11) testing result output function, can will test result output into Excel table, the numerical value of each testing result,
The information such as unit, time and curve graph are all recorded in table.
5th purpose of the invention is to provide a kind of microorganism high-pass culture and the automation of concentration real-time detection is compound
Detector, including the control of above-mentioned Manifold Light Way device, automatic device, composite microporous plate, conductor temperature control device, computer
It further include the light emitting diode matrix of the specific wavelength controlled by computer software except software, wherein
In the lower section of shaking platform 18, light emitting diode matrix light source 50 is installed, and uniform light board 51 is used to make illumination strong
Degree is uniformly distributed below platform, and intensity of illumination and time can be arranged by computer program.
In one embodiment, different combination of light sources may be selected in light emitting diode matrix light source 50 as needed.One
In a preferred embodiment, the light emitting diode of different wavelength range is may be selected in light emitting diode matrix light source 50, small to be suitble to
The high-throughput culture and screening of type algae, photosynthetic bacteria；In a further preferred embodiment, light emitting diode matrix light source 50
According to corresponding two wave-length coverages in each hole in the light emitting diode of 310-1550nm, light genetics research tool can be used as；?
In other preferred embodiments, the light emitting diode of periodical ultraviolet light is may be selected in light emitting diode matrix light source 50, can be to micro-
Biology explores the spectrum of mutation of the ultraviolet radiation as mutagens in genome level, studies ultraviolet radiation in accelerated evolutionary mistake
Effect in journey.
6th purpose of the invention, which is to provide, a kind of carries out microorganism high-pass culture and dense using any of the above-described device
The method of real-time detection is spent, step includes:
(1) temperature controls: the microwell plate 6 for being inoculated with microbiological specimens being covered ventilative upper cover and is put into shaking platform 18, temperature
Degree control device makes the temperature of culturing room inner cavity 24 maintain target temperature in 10 minutes；
(2) shaking culture: shaking platform 18 is driven to do the shaking of high speed circumference with 1-3mm radius by motor 28, shaking is flat
Platform 18 drives microwell plate 6 to shake at a high speed, so that microorganism be made to grow in the fluid nutrient medium with sufficient oxygen；
(3) Concentration Testing: stepper motor 39 drives detection probe 23 to move at a predetermined rate from starting point, successively rests on
First row, secondary series, third column micropore center location, it is every to stop primary, the concentration of one column sample of detection, until having detected all
Sample, stepper motor 39 drive detection probe 23 to return to starting point.
(4) it repeats to detect: using certain time as interval, successively constantly repeating step (3), such motor 28, stepper motor
39 and Concentration Testing pop one's head in 23 threes cooperate, in microbial cultivation process real-time detection concentration change, until experiment knot
Beam.In any of the above-described embodiment, the pipe of light-emitting diodes 1 is light-emitting diodes of the peak wavelength range in 600-1400nm
Pipe, preferably 850nm, microwell plate 6 are that (hole is in 8 × 12 for 24 holes (hole is in 4 × 6 arrangements), 48 holes (hole is in 6 × 8 arrangements) and 96 holes
Arrangement) microwell plate, preferably 48 hole microwell plates.6 height of microwell plate is 20-50mm, preferably 40-45mm.
In any of the above-described embodiment, in measuring each hole of microwell plate 6 when bacteria suspension concentration, light emitting diode 1
It shines, light is transmitted by optical fiber 2 (4), enters bacteria suspension through 6 upper cover of microwell plate, part light is by microbe granular in bacteria suspension
It absorbs or scatters, be not absorbed or the light of forward scattering enters photodiode 9, photodiode 9 will receive optical signal and turn
It is melted into electric signal.Circuit single-chip microcontroller compares built-in calibration curve, obtains bacteria suspension concentration value according to the signal magnitude detected.
In any of the above-described specific embodiment, 28 revolving speed of motor is most fast up to 1000-3000 revs/min, preferably
1500-2000 revs/min, to reach the balance of maximum oxygen transfer ability in 6 micropore of stability and microwell plate for shaking device.
In any of the above-described embodiment, 1 peak wavelength of light emitting diode that the detection method uses for
850nm, the correspondence OD that the detection probe 23 detects600Value range is 0-5OD600, therefore, the present invention can be in 0-5OD600Model
Accurate measurement result is obtained in enclosing.
Detailed description of the invention:
Fig. 1 is the schematic diagram of Manifold Light Way device；
Fig. 2 is the schematic diagram of microwell plate shaking device；
Fig. 3 is shaking platform structure schematic diagram；
Fig. 4 is eccentric shaft structure schematic diagram；
Fig. 5 is the vertical structure schematic diagram of the automation equipment containing shaking driving device；
Fig. 6 is the transverse structure signal of the automation set composite containing shaking driving device and semiconductor temperature-control device
Figure；
Fig. 7 is containing shaking platform, mobile device, the overlooking structure diagram of semiconductor temperature-control device；
Fig. 8 is the hot lid schematic diagram of microwell plate；
Fig. 9 is microwell plate Ventilation cover structure schematic diagram, and longitudinal arrow is path channels；
Figure 10 is the schematic diagram of the automation compound detection instrument containing light emitting diode matrix.
Marginal data:
1: light emitting diode；2: optical fiber；3: optical splitter；4: optical fiber；5: fiber fixed seat；6: microwell plate；7: Liquid Culture
Base；8: photodiode fixing seat；9: photodiode；10: the synchronizing wheel on eccentric shaft；11: synchronous belt；12: bearing is fixed
Plate；13: the synchronizing wheel on motor；14: motor shaft；15: motor fixing plate；16: counterweight；17: eccentric shaft；18: shaking platform；
19: microwell plate fixed station；20: upper bearing (metal) fixed station；21: upper bearing (metal)；22: lower bearing；23: Concentration Testing probe；24: culturing room
Inner cavity；25: heat lid；26: temp probe；27: lower bearing fixed plate support column；28: motor；29: support baseboard；30: fixing seat；
31: fan；32: refrigeration head；33: semiconductor chilling plate；34: cooling fin；35: fan；36: sliding rail；37: positioning rule；38: photoelectricity
Switch；39: stepper motor；40: hot lid shaft；41: heat lid inside heating sheet；42: hot cap bore；43: hot cover lock button hole；44: gold
Belong to top cover；45: hydrophobic permeable membrane；46: light-transmitting plate；47: silicagel pad；48: light-transmitting plate air hole；49: microwell plate hole wall；50: two
Pole pipe array light source；51: uniform light board；
Technical effect:
(1) concentration detection apparatus of the invention using optical splitter and forward scattering optical path in conjunction with mode come stagger the time detect it is micro-
On orifice plate in each hole sample concentration, avoid common multi-channel detection probe simultaneously to multiple sample detections when adjacent holes believe
The shortcomings that number interfering with each other.The work of the optical path by optical splitter, infrared light emission circuit and mid-infrared optical receiving circuit coordination
Control is constituted, and infrared light emission circuit uses dedicated constant-current source chip controls, both can guarantee the use of infrared light-emitting diode
Safety and service life, while also can guarantee the infrared light power stabilising of transmitting.Infrared light emission circuit has light emitting diode guarantor
Protective function, when power supply is reversed or electric current is excessive, light emitting diode can be effectively protected.Mid-infrared optical receiving circuit is using high
Amplifier chip is held, high magnification numbe low noise is built across resistance amplifying circuit, converts optical signals to electric signal.
(2) the high speed shaking platform in instrument and stepper motor cooperate, and high speed shakes liquid, so that culture medium has
Higher oxygen passes to speed and concentration detection apparatus organically combines, and has accomplished full-automation, avoids all of conventional method
More disadvantages.
(3) temperature control equipment in instrument uses semiconductor temperature mode, can be by culture room temperature control in 10-
60 degrees Celsius, it can satisfy the culture from thermophilic cold, room temperature and mesophilic microorganism.
(4) gas exchange device uses metal cover, hydrophobic permeable membrane, light-transmitting plate and silicon with holes inside and outside the microwell plate in instrument
Rubber mat combination is provided for microorganism growth and is filled in addition the hot lid arrangement of microwell plate, can make the gas free exchange inside and outside microwell plate
The oxygen of foot, the liquid excessive evaporation being also avoided that in sample bottle or formation vapor influence testing result.
(5) this self-reacting device is popped one's head in using multi-channel detection, in conjunction with high speed shaking culture and conductor temperature control with
And the technologies such as microwell plate gas exchanges and data acquire, analysis, output software, realize microorganism high-pass culture, concentration
The full-automation of real-time detection and data analysis.Solve high-throughput, real-time concentration detection, microwell plate in current microbe research
Middle culture medium oxygen supply, a wide range of temperature control the problem that cannot be compatible with.The working strength for significantly reducing operator is
Microorganism basic research and bio-engineering research provide a kind of very efficient tool.
Specific embodiment:
Present invention will be further explained below with reference to the attached drawings and examples.But it is as described below, it is only preferable reality of the invention
Example is applied, is not intended to limit the present invention in any form, therefore without departing from the technical solutions of the present invention, according to this
The technical spirit of invention any simple modification, equivalent change and modification to the above embodiments, still fall within skill of the present invention
In the range of art scheme.
As shown in Figure 1, the light path schematic diagram of Concentration Testing probe.Wherein, light source is peak wavelength range in 600-
The light emitting diode 1 of 1400nm, and coaxially coupled with optical fiber 2,2 front end of optical fiber connects optical splitter 3, draws 8-16 from optical splitter 3
Root optical fiber 4, and be fixed on fiber fixed seat 5.Fiber fixed seat 5 is connected with photodiode fixing seat 8 in side, forms U
Shape bracket secures 8-16 photodiode 9 in photodiode fixing seat 8, has high sensitivity, high stability, high line
Property and wavelength of peak sensitivity is within the scope of 1 peak wavelength of light emitting diode.The axis of optical fiber 4 and photodiode 9 is hanging down
Histogram is aligned upwards.There is the gap of 20-50mm between fiber fixed seat and photodiode fixing seat, holds for placing sample
Device, such as microwell plate 6, there is fluid nutrient medium 7 in the micropore of microwell plate 6, and the light that optical fiber issues passes through micropore and fluid nutrient medium
7, the light of forward scattering enters photodiode 9, such light emitting diode 1 and photodiode 9 be formed before to penetrating light inspection
Survey optical path.When detecting, control circuit can make optical splitter 3 successively connect optical fiber 4 with light emitting diode 1, and adjacent optical path is cut
Throw-over degree, without apparent time delay, both ensure that each channel when guaranteeing to carry out multi-channel detection up to Millisecond in this way
The synchronization of detection is not also ensured by the interference of adjacency channel when detection, can almost detect 8-16 channel simultaneously.It is measuring
In micropore when sample concentration, light emitting diode 1 shines according to modulating frequency, and each photodiode 9 receives forward scattering respectively
Optical signal, and direct current signal is exported by pre-amplification circuit, filter circuit, demodulator circuit.
As shown in Fig. 2,3 and 4, the core of microwell plate shaking device is shaking platform driving device and eccentric shaft 17.
Wherein three identical synchronizing wheels 10 are fixed on eccentric shaft 17, and axially eccentric distance is 1-3mm, and eccentric shaft 17 passes through lower axle
It holds 22 to be fixed on the fixed bottom plate 12 of bearing, counterweight 16 is fixed on eccentric shaft 17, the fixed upper bearing (metal) in 17 top of eccentric shaft
21, upper bearing (metal) 21 is fixed on shaking platform 18 by bearing by upper bearing (metal) fixed station 20, and the center of gravity of three counterweights 16 is in level
Direction is in the same direction, and opposite with 17 eccentric direction of eccentric shaft.Motor 28 is fixed on the fixed bottom plate 12 of bearing by clicking fixed plate,
Synchronizing wheel 13 is installed on the axis 14 of motor 28, and is connected with other three synchronizing wheels 10 by synchronous belt 11.Motor 28 is with 1000-
3000 revs/min of revolving speed drives the synchronizing wheel 13 being fixed on motor shaft 14 to rotate, and then drives synchronous belt, and then drives and fix
Synchronizing wheel 10 on eccentric shaft 17 rotates, and then the counterweight 16 on the rotation of eccentric shaft 17 and eccentric shaft 17 is driven to rotate.Due to
There is 1-3mm axially eccentric in 17 top half of eccentric shaft and lower half portion, and (i.e. two inside the eccentric shaft 17 of Fig. 4 are vertical parallel
The axial line distance of line), so 17 top half of eccentric shaft can do circumference shaking by radius of 1-3mm, and then drive shaking platform
18 do circumference shaking by radius of 1-3mm, final to realize that microwell plate 6 does circumference shaking by radius of 1-3mm.Shaking platform 18 can
2-8 24 holes, 48 holes or 96 hole microwell plates 6 are placed, 6 height of microwell plate is 20-50mm.
As illustrated in figs. 5-7, apparatus subject partial internal structure also wraps other than Concentration Testing is popped one's head in and shakes driving device
Include guide rail and semiconductor temperature-control device composition, wherein
U-shaped Concentration Testing probe 23 is fixed on the linear slide rail 36 of the control of stepper motor 39 by fixing seat 30, sliding rail
36 are distributed with shaking platform 18 in vertical equity, and positioning caliper 37 and optoelectronic switch 38 with groove are installed in sliding rail side.Positioning
Stop place when slide calliper rule 37 and the cooperation of optoelectronic switch 38 are for positioning Concentration Testing probe detection.The number of recesses of positioning caliper 37
It is 6-24, each recess edge and the alignment of the microwell plate each column hole center of circle.When Concentration Testing probe slides on the slide rail, microwell plate
6 can pass through among Concentration Testing probe 23.
Conductor temperature control device includes that Peltier semiconductor chilling plate 33, refrigeration head 32, cooling fin 34 and two groups are scattered
Hot-air fan 31,35, refrigeration head 32 are located at 24 inner wall of culturing room inner cavity, and front end connects fan 31, and semiconductor chilling plate 33 is located at
Freeze between head 32 and cooling fin 34, fan 35 is connected on the outside of cooling fin 34.Temp probe 26 on 24 wall of inner cavity measures culture
Indoor temperature simultaneously feeds back to control circuit.
As shown in figure 8, the hot lid 25 of microwell plate is located above microwell plate 6, and with holes 42,26 hole site of 42 position of hole and microwell plate
Corresponding, hot 25 inner band heating sheet 41 of lid can heat microwell plate gas exchange device, it is ensured that the temperature of gas exchange device
It spends 1-2 degrees Celsius higher than 24 temperature of culturing room inner cavity.
As shown in figure 9, inside and outside microwell plate gas exchange device from top to bottom successively when casting coping 44, hydrophobic permeable membrane
45, light-transmitting plate 46 and silicagel pad 47.Casting coping 44, hydrophobic permeable membrane 45 and silicagel pad 47 are all with holes, hole site and microwell plate 6
Hole site is corresponding, passes through convenient for light path, and light-transmitting plate 46 is in corresponding 45 overlay area of hydrophobic permeable membrane band strip-shaped hole 48.
As shown in Figure 10, LED array light source 50 and uniform light board 51, intensity of illumination and time is installed below in shaking platform 18
It can be arranged by computer program.Different wavelength range may be selected in light emitting diode matrix light source, meets the needs of different experiments.
Embodiment one: the method for carrying out microorganism high-pass culture and concentration real-time detection using any of the above-described device
Step includes:
1) fluid nutrient medium and microbial strains to be cultivated are added into microwell plate, cover equipped with hydrophobic permeable membrane and silicon
The upper cover of glue gasket.
2) temperature of culture inner cavity, the i.e. growth temperature of microorganism are determined.Incubator is set on the software of computer in advance
Temperature, sample message, incubation time, Concentration Testing interval time, brushless motor the parameters such as revolving speed.Microwell plate is fixed to
It shakes on platform, then starts detection function on the software of computer, start to test.Culture inner cavity begins to warm up or freezes, with
Reach preset temperature.
3) during the whole culture process, brushless motor continues working, microorganism growth and breeding in microwell plate, when culture is held
After a continuous detection interval time, the concentration of sample in Concentration Testing probe measurement microwell plate after the completion of each detection cycle, is walked
It stops working into motor and Concentration Testing probe, until next detection cycle starts.Each detection cycle, each independent sample
A data, next detection cycle are all produced, each independent sample generates a data again, constantly repeats like this, directly
To after incubation time cut-off, system stalls, the data that each sample generates will form the growth song that the sample corresponds to the time
Line.These individual datas or entire curve can carry out post analysis.
The above described is only a preferred embodiment of the present invention, be not intended to limit the present invention in any form, therefore
Without departing from the technical solutions of the present invention, to the above embodiments according to the technical essence of the invention any simply to repair
Change, equivalent variations and modification, all of which are still within the scope of the technical scheme of the invention.
Claims (10)
1. a kind of Manifold Light Way device of high-throughput detection microbiological specimens concentration, the device include: light emitting diode (1) and
The optical fiber (2) communicated therewith, the optical splitter (3) being connected to optical fiber (2), the multichannel optical fiber (4) being connected to optical splitter (3), with
And Concentration Testing probe (23), wherein
Concentration Testing probe (23) includes as the fiber fixed seat (5) of upper arm and as the photodiode fixing seat of lower arm
(8), fiber fixed seat (5) is connected to above-mentioned multichannel optical fiber (4), and photodiode fixing seat (8) is equipped with and multichannel optical fiber (4)
The corresponding multichannel photodiode (9) of optical path, and upper and lower arms respectively correspond and the vertical upper and lower of microwell plate are arranged in
Side, each other by vertically being connect with the connector of upper lower arm, so that the Concentration Testing for forming the U-shaped opening of horizontal direction is visited
Head；
The optical coupling entering light that light emitting diode (1) issues fixed wave length is fine (2), can be assigned to light after optical splitter (3)
Specified multichannel optical fiber (4), from multiple micropores that a plurality of optical path of optical fiber (4) channel emission vertically penetrates microwell plate (6)
Fluid nutrient medium (7) in sample, and by Concentration Testing probe (23) detection through sample sum by sample scattering before
It is converted to electric signal to scattering light, and by optical signal, then converts electrical signals into digital signal through circuit, whole process is formed
Concentration Testing optical path, to complete the detection of high-flux microorganism sample concentration.
2. the device of claim 1, wherein the upper lower arm of the detection probe (23) fix respectively 8-16 channel optical fiber (4) and
Photodiode (9), the optical fiber (4) and photodiode (9) in preferably 12 channels.
3. the device of claims 1 or 2, wherein the upper lower arm spacing distance of the detection probe (23) is 20-50mm, wherein it is preferred that
It is 40mm；The light emitting diode (1) is peak wavelength range in the light emitting diode of 600-1400nm, preferably 850nm.
4. a kind of automation equipment of microorganism high-pass culture and concentration real-time detection, the preceding claims 1 including top
To Manifold Light Way device described in 3, the shaking platform of the multiple microwell plates of carrying at middle part, the microwell plate of lower part shakes device,
And it is located at the detection probe mobile device of bottom, wherein
(1) shaking platform (18) is located between the U-shaped opening of Concentration Testing probe, and Concentration Testing probe can be on shaking platform
Multiple microwell plate upper horizontals are mobile；
(2) microwell plate being fixedly connected with shaking platform (18) shakes device, and the motor (28) including bottom is located at motor (28)
The bearing (setting) plate (12) of top, three eccentric shafts (17) of asymmetric setting, each eccentric shaft (17) on bearing (setting) plate (12)
Middle section is connected with each other by synchronous belt (11), and eccentric shaft (17) upper section is fixedly connected with shaking platform (18), and synchronous belt (11) twines
On machine shaft (14) in fixed plate, when motor (28) driving motor shaft (14) rotation, shaft (14), which drives, to be synchronized
Band drives multiple eccentric shaft (17) rotations, to drive top shaking platform (18) high speed circumference shaking, finally drives microwell plate
(6) high speed circumference shakes；
(3) detection probe mobile device includes the linear slide rail (36) of stepper motor (39) control, the sliding rail and shaking platform
(18) it is distributed in vertical equity, one end is connected with stepper motor (39), and the other end passes through fixing seat fixing seat (30) and top
Upper arm or the lower wall connection of Concentration Testing probe (23) can drive the dense of top when stepper motor (39) driving sliding rail is mobile
The U-shaped opening for spending detection probe (23) is moved from the vertically side of the microwell plate of shaking platform.
5. the automation equipment of claim 4, wherein eccentric shaft (17) is equipped with synchronizing wheel in microwell plate shaking device
(11), it and by synchronous belt (11) is connected with each other；Motor (28) is fixed on motor fixing plate (15) by flange, and motor is fixed
Plate (15) is fixed on bearing (setting) plate (12), and motor fixing plate (15) is equipped with machine shaft (14), and solid with synchronizing wheel (13)
Fixed connection, so that motor (28) is connected by synchronizing wheel (13), synchronous belt (11) with the synchronizing wheel (11) of three eccentric shafts (17).
6. the automation equipment of claim 4-5, wherein three eccentric shaft (17) lower ends are fixed lower bearing (22), lower bearing (22)
It is fixed on bearing (setting) plate (12), eccentric shaft (17) upper end is fixed upper bearing (metal) (21), and upper bearing (metal) (21) is fixed to shaking platform
(18) on, wherein the axis of the circular motion of eccentric shaft upper and lower ends is in two parallel lines of spacing 1-3mm, synchronous pulley (13)
It is fixed on eccentric shaft (17) lower end with counterweight (16), counterweight (16) center of gravity direction and eccentric shaft (17) offset direction are in the horizontal direction
It is exactly the opposite, with counteracting or partial offset because shaking platform bias shakes the uneven centrifugal force to be formed.
7. the automation equipment of claim 4-6, wherein in the mobile device, positioning with groove is installed in sliding rail (36) side
Slide calliper rule (37), fixed concentration detection probe (23) and the optoelectronic switch for converting optical signals to electric signal on sliding rail slide unit
(38), stop place when positioning caliper (37) and optoelectronic switch (38) cooperation are for positioning Concentration Testing probe (23) detection.
8. the automation equipment of claim 4-7, wherein shaking platform (18) can place 2-8 microwell plate, in shaking platform
(18) it is arranged in longitudinal 2 × 1,2 × 2,2 × 3,2 × 4, preferably 2 × 2 arrangements；Used microwell plate (6) is that (hole is in 24 holes
4 × 6 arrangements), 48 holes (hole in 6 × 8 arrangement) and 96 holes (hole is arranged in 8 × 12) microwell plate (6), preferably 48 hole microwell plates
(6)；The microwell plate (6) is highly 20-50mm, preferably 40mm.
9. the automation equipment of claim 4-8, wherein motor (28) revolving speed is 1000-3000 revs/min, preferably 1500-
2000 revs/min, to reach the balance of maximum oxygen transfer ability in the stability and micropore of shake device.
10. the automation equipment of claim 4-9, wherein the number of recesses of positioning caliper (37) is a for 6-24, and preferably 16,
Each recess edge and the alignment of microwell plate (6) each column hole center of circle.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112730383A (en) * | 2020-12-28 | 2021-04-30 | 西安交通大学 | Optical fiber array LIBS detection system for online detection |
| WO2022128809A3 (en) * | 2020-12-14 | 2022-09-01 | Qinstruments Gmbh | Laboratory apparatus comprising a mixing mechanism for mixing a medium of a slide and method for mixing a medium in a slide |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20110000463A (en) * | 2009-06-26 | 2011-01-03 | (주)비엔피테크 | Multi-channel photobioreactor for culturing photosynthetic microorganisms |
| CN103121008A (en) * | 2012-06-07 | 2013-05-29 | 上海智城分析仪器制造有限公司 | Planar linkage cantilever type reciprocating oscillation mechanism of culturing oscillator |
| CN105044038A (en) * | 2015-08-31 | 2015-11-11 | 广州中国科学院先进技术研究所 | Non-invasive biomass online detection device used for shake-flask culture |
| CN105907636A (en) * | 2016-05-20 | 2016-08-31 | 长沙湘资生物科技有限公司 | Automatic instrument for microorganism high-flux culture and large concentration range real-time detection |
| CN106990086A (en) * | 2017-06-06 | 2017-07-28 | 中国人民大学 | A kind of multichannel Evanescent Wave All-fiber biology sensor |
| CN108693079A (en) * | 2018-04-16 | 2018-10-23 | 东南大学 | Particle flow parameter measuring apparatus and method based on laser backscatter method |
-
2019
- 2019-01-28 CN CN201910082576.XA patent/CN109554292A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20110000463A (en) * | 2009-06-26 | 2011-01-03 | (주)비엔피테크 | Multi-channel photobioreactor for culturing photosynthetic microorganisms |
| CN103121008A (en) * | 2012-06-07 | 2013-05-29 | 上海智城分析仪器制造有限公司 | Planar linkage cantilever type reciprocating oscillation mechanism of culturing oscillator |
| CN105044038A (en) * | 2015-08-31 | 2015-11-11 | 广州中国科学院先进技术研究所 | Non-invasive biomass online detection device used for shake-flask culture |
| CN105907636A (en) * | 2016-05-20 | 2016-08-31 | 长沙湘资生物科技有限公司 | Automatic instrument for microorganism high-flux culture and large concentration range real-time detection |
| CN106990086A (en) * | 2017-06-06 | 2017-07-28 | 中国人民大学 | A kind of multichannel Evanescent Wave All-fiber biology sensor |
| CN108693079A (en) * | 2018-04-16 | 2018-10-23 | 东南大学 | Particle flow parameter measuring apparatus and method based on laser backscatter method |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022128809A3 (en) * | 2020-12-14 | 2022-09-01 | Qinstruments Gmbh | Laboratory apparatus comprising a mixing mechanism for mixing a medium of a slide and method for mixing a medium in a slide |
| CN112730383A (en) * | 2020-12-28 | 2021-04-30 | 西安交通大学 | Optical fiber array LIBS detection system for online detection |
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