CN103323433B - The shaking table device of contactless multichannel on-line checking fermentation liquid and method of testing - Google Patents
The shaking table device of contactless multichannel on-line checking fermentation liquid and method of testing Download PDFInfo
- Publication number
- CN103323433B CN103323433B CN201310246480.5A CN201310246480A CN103323433B CN 103323433 B CN103323433 B CN 103323433B CN 201310246480 A CN201310246480 A CN 201310246480A CN 103323433 B CN103323433 B CN 103323433B
- Authority
- CN
- China
- Prior art keywords
- microorganism
- shaking table
- laser
- optical density
- test
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Abstract
The present invention provides shaking table device and the method for testing of a kind of contactless multichannel on-line checking fermentation liquid, the shaking table device of multichannel on-line checking fermentation liquid is, n the test bottle filling fermentation liquid it is provided with on shaking table, each test bottle side is provided with generating laser, opposite side is provided with laser pickoff, the output of laser pickoff is connected with data acquisition and processing (DAP) module, data acquisition and processing (DAP) module is connected with computer, constitute n TCH test channel, computer is by n the microorganism growth process filling different fermentations liquid, it is depicted as curve respectively, the growth course of microorganism in each test bottle of monitoring in real time.One section of cylinder is arranged at the bottom of described test bottle.Advantage it is possible to be illustrated by curve, each process of growth of microorganism in each test bottle on shaking table directly perceived, in real time, conveniently, substantially increases scientific research and microniological proudcts production efficiency.
Description
Technical field
The present invention relates to biochemical instrument technical field, particularly relate to a kind of in shaking table device, arrange n fill fermentation
The test bottle of liquid, monitors in real time device and the method for testing of microorganism growth process in each test bottle.
Background technology
At present, fermentation liquid microorganism growth process in test bottle on domestic monitoring shaking table, is all to use off-line test method.
Way is to fermentation liquid timing sampling in single test bottle, is transplanted in spectrophotometer cell, then tests its light transmittance
With optical density (spectrophotometer is commonly referred to as absorbance A), measured values is drawn with time relationship.This method is complete
Manual operations, it is impossible to on-line real-time measuremen, the most very time-consuming.
Summary of the invention
For solving the problem of aforementioned existence, the present invention provides the shaking table of a kind of contactless multichannel on-line checking fermentation liquid
Device and method of testing, the shaking table device of multichannel on-line checking fermentation liquid is, shaking table is provided with n the survey filling fermentation liquid
Trial jar, each test bottle side is provided with generating laser, and opposite side is provided with laser pickoff, the output of laser pickoff with
Data acquisition and processing (DAP) module connects, and data acquisition and processing (DAP) module is connected with computer, constitutes n TCH test channel, computer
By n the microorganism growth process filling different fermentations liquid, it is depicted as curve respectively, monitors microorganism in each test bottle in real time
Growth course.
One section of cylinder is arranged at the bottom of described test bottle.
The method of testing of the shaking table device of multichannel on-line checking fermentation liquid is:
(1) numbering of n passage is set, presets n passage maximum optical density value, start shaking table 50-250rps；
(2) when Laser Measurement is non-lit up, the n times meansigma methods of the minimum output voltage value of the laser pickoff of each passage；
(3) generating laser is lighted；
(4) the n times meansigma methods of the maximum voltage value of the output of the laser pickoff measuring n passage is gathered；
(5) the real-time voltage value of n multichannel laser receptor output of measurement is gathered；
(6) light transmittance of microorganism in calculating test bottle；
Light transmittance=((real-time voltage value-minimum voltage meansigma methods)/(maximum voltage meansigma methods-voltage is little averagely
Value)) × 100；
(7) the optical density OD value of microorganism in calculating test bottle；
Optical density (OD)=LOG(maximum transmission rate/light transmittance)；
Maximum transmission rate=100%；
(8) in drawing each lane testing bottle, microorganism optical density changes over curve；
(9) reach to preset experimental period or optical density value, off-test, preserve data；
(10) not up to preset experimental period or optical density value, return step (5).
The optical maser wavelength of generating laser output typically selects 600nm or 660nm, because the light wave many at this wavelength is micro-
Biology itself does not absorb luminous energy, only reflects or scatters light wave.Can also be different according to microorganism property, select swashing of other wavelength
Luminous source, so, this experiment is the turbidity determining microorganism, but microbe research personnel get used to optical density OD table
Show, be identical physical concept with the absorbance A of spectrum analysis.
Optical density does not has dimensional unit, is a logarithm value, optical density be incident illumination with the logarithm of transmission light ratio or
Say it is the reciprocal logarithm of light penetration.Computing formula is OD=log10(incident illumination/transmission light) or OD=log10(1/ printing opacity
Rate)；
The optical maser wavelength of generating laser output typically selects 600nm or 660nm, because the light wave many at this wavelength is micro-
Biology itself does not absorb luminous energy, only reflects or scatters light wave.Can also be different according to microorganism property, select swashing of other wavelength
Luminous source, so, this experiment is the turbidity determining microorganism, but microbe research personnel get used to optical density OD table
Show, be identical physical concept with the absorbance A of spectrum analysis.
It is an advantage of the invention that and can be illustrated by curve, growth of microorganism each in each test bottle on shaking table directly perceived
Individual process, in real time, conveniently, can provide the experimental provision of automatization, also can produce for microniological proudcts and provide for microbe research
Optimize data and automatically control information.Substantially increase scientific research and microniological proudcts production efficiency.
Accompanying drawing explanation
Accompanying drawing 1 is microorganism fungus kind increment curve.
Accompanying drawing 2 is block diagram of the present invention.
Accompanying drawing 3 is test bottle axonometric chart of the present invention.
Accompanying drawing 4 is the schematic diagram (arrow signal laser light test bottle in figure) having fermentation liquid in test bottle of the present invention.
Accompanying drawing 5 is laser diode driver circuit figure.
Accompanying drawing 6 shows it is switching regulator laser diode driver circuit figure.
Accompanying drawing 7 is three kinds of fermentation process optical density curve figures.
Detailed description of the invention
Referring to shown in accompanying drawing 2, shaking table is provided with n the test bottle 2 filling fermentation liquid, each test bottle 2 side sets
Being equipped with generating laser 1, generating laser 1 uses laser diode, and wavelength is 600nm(or 660nm)；Opposite side is provided with sharp
Optical receiver 3, uses silicon photoelectric diode, and wavelength is 550 750nm or joins narrow band pass filter；The output of laser pickoff 3 with
Data acquisition and processing (DAP) module 4 connects, and data acquisition and processing (DAP) module 4 is connected with computer 5, constitutes n TCH test channel, calculates
Machine 5, by n the microorganism growth process filling different fermentations liquid, is depicted as curve respectively, micro-in monitoring each test bottle in real time
Biological growth course.
In biotechnology, shaking table is research tool, is also the tool of production.At growth of microorganism sweat and
Biological medicine development aspect needs to monitor the various chemical-physical parameters change of sample, and wherein, spectrodensitometry is exactly one
Emphasis, because it is the change of Biomass in monitoring biological growth sweat, shows and judges each stage of biological respinse
Feature, seek optimal biochemical reaction pattern and optimize industry mathematical model important evidence；Such as shown in accompanying drawing 1, microorganism
Sweat has four-stage, and in figure, label is:
Viable count ... total bacteria count
A lag phase；B increased logarithmic phase；
C stable phase；D decline phase；
In scientific research, by the concentration change in monitoring growth of microorganism sweat in real time, just can sentence and distinguish micro-life
In stage residing for thing fermentation, seek biochemical reaction route.
On producing, in order to improve productivity, in conjunction with the measurement of other the physical-chemical parameters, set up and optimize biochemical reaction mathematics
Model, in order to instruct and to control production process.
Biological product are not have absorption spectrum at 600nm or 660nm at wavelength mostly, so generally using this most individual ripple
The optical density of long detection microorganism.When working due to shaking table, it is in ceaselessly motion (vibrate or rotate), therefore to optical density
Measurement adds difficulty: the most under vibrating conditions, it is ensured that the mechanical stability of whole optical system;2. container tube wall pair is reduced
The refraction of incident light source；3. liquid fluctuating in container is reduced to the reflection of light and refraction；4. overcome fluid to be measured inner cell micro-
The grain scattering to light；5. the focusing of light source device, power selection and temperature stability control；6. the stablizing of electrooptical device
Property and the sensitivity uniformity of photosurface；So in such a case can long-term work in real time, have stable, accurate, repeated
Good optical density test equipment is very important.
To this end, when using shaking table carry out Fermentation Experiment or produce, general employing commodity conical flask holds cultivation
Liquid.In order to ensure microorganism dissolved oxygen amount in growth sweat, can only add in bottle a small amount of culture fluid (30% with
In).Thus there are the following problems: 1. in order to measure the optical density (OD600 or OD660) of culture fluid, is necessary for laser light
The horizontal position in source is put the lowest, and otherwise in liquid level constantly fluctuates, light will escape liquid level, causes change in optical path length, produces
Detection signal fluctuation.2. too low when laser light source horizontal position, light it is possible to be in the surface location at the bottom of conical flask, then when
After light, through refraction or the scattering of this position glass, defocus, pass to detector end face and just become a bar shaped
Speckle.If 3. adding sample, in sample container in rocking, liquid fluctuating can be produced, just as mirror, light
Scattering four directions, this strip light spots more to expand, and beyond the receiving plane of detector, causes bigger detection signal fluctuation.Therefore,
For above-mentioned problem and shortage, the present invention have developed a kind of Novel tapered bottle.Fig. 3 is the structure of conical flask of the present invention, in figure
One section of cylinder is arranged at the bottom of bottle, and laser passes through from this cylinder, will not produce the refraction of light.Fig. 4 is to have a small amount of cultivation in bottle
Liquid, laser passes through from the bottom culture fluid of bottle, and laser is not reflected or dispersion effect by this Novel tapered bottle.
Generating laser is the light source of a branch of single wavelength, utilizes micro lens to focus on, from light source outlet 100mm's
Focus diameter is that 2-4mm is adjustable；Laser beam microbiological specimens in conical flask absorbs, stops or scatter, and finally arrives silicon light
The luminosity of electric diode 3 is converted into electric current.
If laser luminosity=Iin
By light-metering luminosity=Iout
Absorbed light=Ia
Scattered light and the luminosity=Id of light being blocked
Therefore Iout=Iin-(Ia+Id)
Select the laser of 600nm and 660nm wavelength as exciting light, then Ia=0, then stopped by microorganism or scatter
Luminosity is Id.According to photometer computing formula,
OD=Log (Iout/Iin) light path is 10mm
In view of the impact of light path, OD=f(Iout, Iin, S) rate
S light path
In general spectroscopic instruments, light path is mostly 10mm.And the light path of monitoring shaking table in real time is relevant to shaking flask capacity, with
250mL shaking flask is as the criterion, and its light path is about 100mm, and this will increase excitation luminous power 10 times.And increase power and bring laser tube
Stablizing of heating, minimizing persistently use time and luminosity.Therefore, it is necessary to select the laser drive circuit with power stability.Or
Person, uses on-off control LASER Light Source.Fig. 5 is the laser diode drive schematic diagram of power stability；Fig. 6 switching regulator laser
Diode drive circuit.
Triangular flask is not only microbial reaction container and has been become an optics in whole optical system, typically
There is a chamfering curved surface bottom commercial triangular flask, be exactly in incident beam into and out of region, the refraction to light beam of this curved surface,
When adding shaking table vibration, the reflection of microorganism liquid level or refraction, will cause photoelectric detecting signal seriously to be beated, and Fig. 3,4 be
The test bottle that invention improves, has one section of cylinder bottom bottle, laser passes through at cylinder, will not produce the refraction of light.
Laser pickoff, uses silicon photoelectric diode, and its dark current is little, range of linearity width, good stability；In order to avoid
Additional detection voltage drift, the equipment circuit that only one performance of need is good follows output；And in order to prevent shaking table work tool on opportunity
The interference of vibration, selects detection surface area big, the uniform silicon photoelectric diode of light sensitivitys of photosurface.
Data acquisition unit at least to select the A/D harvester of more than 12 resolution.During shaking table work, due to microorganism base
Liquid fluctuates in container, causes optical density to detect signal fluctuation, therefore can not use high-speed AD circuit, and preferably use double integrator or
Delta-cgma low speed analog-digital converter (currently employed MSP 020102,24 bit analog-digital converter)；
Concrete result of the test:
It is further elucidated with summary of the invention below in conjunction with embodiment, but these examples are not limiting as the protection model of the present invention
Enclose.
1) shaking table is selected: Zhi Cheng analytical tool Manufacturing Co., Ltd production ZWY-100H
2) temperature: 37 DEG C
3) rotating speed: 150 240rps
4) microbiological specimens: escherichia coli；Agrobacterium and Shanghai Communications University's Onlly microbiological specimens
5) culture medium: unified use identical goods culture medium
6) port number: 1-4
For optimizing the growth fermentation condition of microorganism, choose suitable culture medium, use apparatus of the present invention to pass through triple channel
Cultivate three microorganism fungus kinds: escherichia coli, Agrobacterium and Onlly strain.Incubation time is 20 hours, three growth
The real-time monitoring result of optical density in sweat, through data acquisition and transport module collection and be transferred to terminal, enters
Row Data Analysis Services, finally obtains microorganism densitometric data of real-time growth sweat in culture fluid and is shown in Table 1, paint
The optical density of system changes over curve and sees Fig. 7.
Optical density change (automatically generating Excel electronic watch) in 1 three kinds of microorganism fungus kind sweats of table:
| Time (divides) | Passage 2 | Passage 3 | Passage 4 |
| 11.4 | 0 | 0.003 | -0.001 |
| 47.4 | 0 | 0.019 | -0.001 |
| 83.5 | -0.001 | 0.047 | -0.001 |
| 95.5 | -0.001 | 0.06 | -0.001 |
| 131.5 | -0.001 | 0.111 | -0.001 |
| 167.6 | -0.001 | 0.19 | -0.001 |
| 203.6 | -0.001 | 0.313 | -0.001 |
| 239.7 | 0 | 0.501 | -0.001 |
| 275.8 | 0.001 | 0.754 | -0.001 |
| 311.8 | 0.047 | 1.031 | -0.001 |
| 347.9 | 0.334 | 1.27 | -0.001 |
| 383.9 | 0.719 | 1.405 | 0.001 |
| 420 | 1.034 | 1.502 | 0.033 |
| 456 | 1.207 | 1.586 | 0.203 |
| 492.1 | 1.324 | 1.644 | 0.471 |
| 528.1 | 1.416 | 1.695 | 0.8 |
| 564.2 | 1.494 | 1.738 | 1.124 |
| 600.2 | 1.557 | 1.776 | 1.312 |
| 636.3 | 1.611 | 1.801 | 1.424 |
| 672.3 | 1.659 | 1.827 | 1.513 |
| 708.4 | 1.706 | 1.848 | 1.582 |
| 744.4 | 1.749 | 1.874 | 1.64 |
| 780.5 | 1.785 | 1.896 | 1.687 |
| 816.5 | 1.819 | 1.921 | 1.73 |
| 852.6 | 1.852 | 1.942 | 1.77 |
| 888.7 | 1.877 | 1.952 | 1.803 |
| 924.7 | 1.905 | 1.965 | 1.836 |
| 960.8 | 1.917 | 1.966 | 1.858 |
| 996.8 | 1.927 | 1.96 | 1.875 |
| 1032.9 | 1.938 | 1.963 | 1.892 |
| 1068.9 | 1.926 | 1.927 | 1.883 |
| 1105 | 1.926 | 1.92 | 1.89 |
| 1141 | 1.932 | 1.918 | 1.899 |
| 1177.1 | 1.94 | 1.922 | 1.912 |
| 1201.1 | 1.94 | 1.921 | 1.918 |
This experimental result has clearly demonstrated that optical density change that microorganism grows in sweat on shaking table (that is micro-life
Substrate concentration changes), reflect the feature of the Main Stage of fermentable well, provide guidance for research and production and borrow
Mirror effect, also simplify operating process simultaneously, and saving experimenter needs tens hours timing samplings (within general one hour, to gather sample
The most once), on spectrophotometer, OD600 is then tested.Which not only improves work efficiency, decrease the work of research worker
Intensity, it is of special importance that optimized mathematical model can be provided for biotechnology research achievement industrialization, for continuous fermentable
Or automatically controlling of biochemical reaction lays the foundation.
Claims (1)
1. a shaking table device detection method for contactless multichannel on-line checking fermentation liquid, shaking table is provided with n and fills
The test bottle of fermentation liquid, each test bottle side is provided with generating laser, and opposite side is provided with laser pickoff, laser pick-off
The output of device is connected with data acquisition and processing (DAP) module, and data acquisition and processing (DAP) module is connected with computer, constitutes n test logical
Road, computer, by n the microorganism growth process filling different fermentations liquid, is depicted as curve respectively, monitors each test in real time
The growth course of microorganism in bottle, detection method comprises the following steps:
(1) numbering of n passage is set, presets n passage maximum optical density value, start shaking table 50-250rps；
(2) when Laser Measurement is non-lit up, the n times meansigma methods of the minimum output voltage value of the laser pickoff of each passage；
(3) generating laser is lighted；
(4) the n times meansigma methods of the maximum voltage value of the output of the laser pickoff measuring n passage is gathered；
(5) the real-time voltage value of n multichannel laser receptor output of measurement is gathered；
(6) light transmittance of microorganism in calculating test bottle；
Light transmittance=((real-time voltage value-minimum voltage meansigma methods)/(maximum voltage meansigma methods-the little meansigma methods of voltage)) ×
100；
(7) the optical density OD value of microorganism in calculating test bottle；
Optical density (OD)=LOG(maximum transmission rate/light transmittance)；
Maximum transmission rate=100%；
(8) in drawing each lane testing bottle, microorganism optical density changes over curve；
(9) reach to preset experimental period or optical density value, off-test, preserve data；
(10) not up to preset experimental period or optical density value, return step (5).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310246480.5A CN103323433B (en) | 2013-06-20 | 2013-06-20 | The shaking table device of contactless multichannel on-line checking fermentation liquid and method of testing |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310246480.5A CN103323433B (en) | 2013-06-20 | 2013-06-20 | The shaking table device of contactless multichannel on-line checking fermentation liquid and method of testing |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103323433A CN103323433A (en) | 2013-09-25 |
| CN103323433B true CN103323433B (en) | 2016-08-10 |
Family
ID=49192318
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310246480.5A Active CN103323433B (en) | 2013-06-20 | 2013-06-20 | The shaking table device of contactless multichannel on-line checking fermentation liquid and method of testing |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103323433B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102519898A (en) * | 2011-12-29 | 2012-06-27 | 上海智城分析仪器制造有限公司 | Device utilizing single light source to detect fermentation liquid of shake table |
| CN102539366A (en) * | 2011-12-29 | 2012-07-04 | 上海智城分析仪器制造有限公司 | Device for detecting swinging bed fermentation liquid by double light sources |
| CN202415551U (en) * | 2011-12-29 | 2012-09-05 | 上海智城分析仪器制造有限公司 | Shake flask for photoelectric detection of fermentation liquor on rocking device |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9880126B2 (en) * | 2010-09-24 | 2018-01-30 | Ajou University Industry-Academic Cooperation Foundation | Biosensor based on carbon nanotube-electric field effect transistor and method for producing the same |
-
2013
- 2013-06-20 CN CN201310246480.5A patent/CN103323433B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102519898A (en) * | 2011-12-29 | 2012-06-27 | 上海智城分析仪器制造有限公司 | Device utilizing single light source to detect fermentation liquid of shake table |
| CN102539366A (en) * | 2011-12-29 | 2012-07-04 | 上海智城分析仪器制造有限公司 | Device for detecting swinging bed fermentation liquid by double light sources |
| CN202415551U (en) * | 2011-12-29 | 2012-09-05 | 上海智城分析仪器制造有限公司 | Shake flask for photoelectric detection of fermentation liquor on rocking device |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103323433A (en) | 2013-09-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Masrie et al. | Detection of nitrogen, phosphorus, and potassium (NPK) nutrients of soil using optical transducer | |
| Ramane et al. | Detection of NPK nutrients of soil using Fiber Optic Sensor | |
| CN102798602B (en) | Integrated dual-beam water quality COD on-line detection sensor | |
| CN101975764B (en) | Near infrared spectrum technology-based multiband soil nitrogen detection device and method | |
| CN104730054A (en) | Integrated probe type photoelectric water-quality multi-parameter online measuring system | |
| CN104777108B (en) | The detection means and method of a kind of chlorophyll content | |
| CN102539378A (en) | Semiconductor laser array near infrared spectrometer | |
| CN204536203U (en) | A kind of integrated probe formula photoelectricity multi-parameter water-quality on-line measurement system | |
| CN103175634A (en) | TDLAS (tunable diode laser absorption spectroscopy) temperature calibration system | |
| CN105548128A (en) | Method and device for detecting chlorophyll of coastal zone water body in situ through double optical path method | |
| CN104062265A (en) | Detection device and detection method for multi-component gas in transformer oil based on spectrum analysis | |
| CN101975759A (en) | Transmission-type nondestructive measuring device and method of water content of plant leaves | |
| CN104198388A (en) | Online water quality monitoring device based on composite spectrum measurement | |
| CN103884706A (en) | Online original pulp liquor detection and classification system | |
| CN103884705A (en) | Original pulp liquor detection and subpackage device | |
| CN104155247A (en) | In-situ measurement method and device for chlorophylls and turbidity of water body | |
| CN101566564A (en) | Detecting system and method for detecting ethanol concentration by reflecting interference spectrum | |
| CN105842178A (en) | Non-intrusive mode online detection device for pH value of cell culture fluid and detection method | |
| CN101929952A (en) | Air quality continuous on line monitor gas calibrating method and device | |
| Jiménez-Márquez et al. | High-resolution low-cost optoelectronic instrument for supervising grape must fermentation | |
| CN103323433B (en) | The shaking table device of contactless multichannel on-line checking fermentation liquid and method of testing | |
| CN205404410U (en) | Double -light -path method littoral zone water chlorophyll normal position monitoring devices | |
| CN204128968U (en) | Original plasm wine on-line checkingi hierarchy system | |
| CN206192871U (en) | Online full gloss register for easy reference water quality analyzer | |
| JP5774352B2 (en) | A non-contact method for continuous measurement of sample growth during culture |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |