CN1289918A - Method and device for multi-channel spectrophotometric detection of rapid microscope - Google Patents
Method and device for multi-channel spectrophotometric detection of rapid microscope Download PDFInfo
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- CN1289918A CN1289918A CN 00117473 CN00117473A CN1289918A CN 1289918 A CN1289918 A CN 1289918A CN 00117473 CN00117473 CN 00117473 CN 00117473 A CN00117473 A CN 00117473A CN 1289918 A CN1289918 A CN 1289918A
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Abstract
A high-speed multi-channel spectrophotometric detector is composed of microscope, photosplitter, multi-channel spectrometer, synchronous data acquisition card, and computer. Its detection method includes spreading the specimen light from microscope onto multi-channel spectrometer by splitter to dectect the light intensities at different wavelengthes, outputting the photoelectric signals to computer in time order by data acquisition card, processing the signals by computer to obtain the absorption spectra at different wavelengthes. Its advantages include high resolution, short time and convenient operation.
Description
The present invention relates to photoelectron and spectral technique, particularly a kind of wavelength resolution height, sample time is short, sample volume is little quick multi channel micro-spectrophotometry detection method and device thereof.
The microspectrophotometry detection technique is a kind of optical microscope and spectrophotometer to be combined, and composition carries out qualitative, quantitatively or the advanced technology analyzed of local positioning in the pair cell on the intact cell level.It is advanced to qualitative cytochemical methods the research range of the small Local Quantitative that quantizes.Utilize this technology to carry out photo densitometry, spectral absorption measurement, fluorophotometric measurement to biological tissue, cell, cellular products and bacterium, parasite etc.; Because the metabolism composition of research object is very complicated, come in every shape, just can tell small variation so need sufficiently high wavelength resolution; In addition, a lot of samples are the living cells sample especially, its absorption spectrum may be a change procedure with the process of life, so the fast detecting of absorption spectrum also is very important, so, both needed sufficiently high wavelength resolution and also needed sufficiently high temporal resolution simultaneously for the microspectrophotometry detection technique.Existing microspectrophotometry detection technique mainly contains two classes: a kind of is to adopt single channel optical detection system, is write down the absorption light intensity of each wavelength after the spectrum of absorption of sample is by the monochromator beam split by detector.Since generally will in 350～800nm scope one by one wavelength detects, so grow (generally being longer than 1 minute) detection time, be difficult to carry out the spectral detection that time resolution is had relatively high expectations, and instrument is bulky, complex structure, it is inconvenient to operate; Another kind is the novel multi channel micro-spectrophotometry detection system of developing in recent years.This system uses diode array as detecting device, can make to add detection time near 20ms, and spectral wavelength sensing range 350nm～800nm, minimum detection zone 5 μ m, but because it adopts diode array, spatial resolution is very limited.Therefore, the wavelength resolution of this type of microspectrophotometry detection system is only in the 2.5nm magnitude.
The objective of the invention is to overcome the shortcoming of above-mentioned prior art, a kind of wavelength resolution height, short quick multi channel micro-spectrophotometry detection method of sampling and measuring time are provided.
Another object of the present invention is to overcome the shortcoming of above-mentioned prior art, a kind of device that can realize said method is provided.
This fast multi channel micro-spectrophotometry detection method comprise following operation steps: the optical convergence that (1) microscope will transmit from sample to the optical splitter after, compose on the multiple tracks spectroscopic detector by the optical splitter exhibition; (2) each sensitization pixel of multiple tracks spectroscopic detector detects the light intensity of each wavelength; (3) the synchronous data collection card outputs to computing machine chronologically with the photoelectron signal of each sensitization pixel of multiple tracks spectroscopic detector; (4) computing machine deals with the absorption spectrum that the back obtains each wavelength to photoelectron signal.
Described computing machine to the process that photoelectron signal deals with is: adopt the sample averaging method to reduce the noisy at random of multiple tracks spectroscopic detector earlier, improve its signal to noise ratio (S/N ratio), then with the Fast Median Filtering method, make it to realize quick high s/n ratio sampling, at last with the absorption light intensity (or transmitted light intensity) of each wavelength correspondence of analytical calculation computed in software and with form or the output of curve image.
Realize that the device of this quick multi channel micro-spectrophotometry detection method is made up of microscope, optical splitter, multiple tracks spectroscopic detector, synchronous data collection card, computing machine, the transmitted light of microscope detection sample is received by the multiple tracks spectroscopic detector after optical splitter exhibition spectrum, the output terminal of multiple tracks spectroscopic detector and synchronous data collection card are electrically connected, and the output terminal and the computing machine of synchronous data collection card are electrically connected.
Quick multi channel micro-spectrophotometry detection method of the present invention and device thereof have following advantage and beneficial effect with respect to prior art: (1) the present invention can directly be that the microcell sample of 1 μ m～125 μ m carries out quick spectral detection to line, and can reach soon≤1ms detection time; In other words, can provide more than 1,000 a full spectrum absorption spectrum in 1 second, and wavelength resolution can reach≤0.2nm, institute's photometry spectrum wavelength scope is decided according to the multiple tracks spectroscopic detector of selecting for use; (2) characteristics of survey fast imaging of spectrum, resolution height in the wavelength gamut owing to having, integral time is adjustable, signal to noise ratio (S/N ratio) is high, volume is little, being convenient to operate, and system software under the Windows operating environment and the powerful data-handling capacity of computing machine, so utilize the present invention can realize fast, accurately, multi-functional microscopic sample analysis ability such as unperturbed, multispectral micrometering, thereby the present invention has than existing other microspectrophotometry technology remarkable advantages and obvious improvement more.
The present invention is described in further detail below in conjunction with accompanying drawing.
Fig. 1 is the structural representation of the quick multi channel micro-spectrophotometry pick-up unit of the present invention.
Embodiment 1:
As seen from Figure 1, the quick multi channel micro-spectrophotometry pick-up unit of the present invention comprises assemblies such as microscope, optical splitter, multiple tracks spectroscopic detector (line array CCD), synchronous data collection card, computing machine; Wherein microscope adopts TE300 type (or the Olympus IX70 type) inverted microscope of Nikon, optical splitter adopts the reflection grating of 1200 lines/mm, the multiple tracks spectroscopic detector adopts the line array CCD of TCD1206 CCD, this line array CCD has the synchronous data collection card simultaneously, the synchronous data collection card is electrically connected formation successively by A/D converter, static memory, address production electric circuit, data bus interface circuit (isa bus) and A/D startup, data read-write control circuit etc., and computing machine can adopt the brand name computer or the compatible of band PIICPU and 64M internal memory and 2G hard disk.Can adopt microscope Halogen lamp LED or fluorescence halogen lamp as light source during microscope detection sample, microscope detects behind the light of sample area transmission by object lens and diaphragm, received to line array CCD by reflection grating exhibition spectrum, each sensitization pixel of high speed linear array CCD detects the light intensity of each wavelength.The output terminal of line array CCD and synchronous data collection card are electrically connected, the high-speed synchronous data acquiring card outputs to the computing machine that is attached thereto chronologically with the photoelectron signal of each sensitization pixel of CCD, computing machine is earlier by a Control Software of operating under the Windows environment, adopt the sample averaging method to reduce the noisy at random of CCD multichannel detector, improve its signal to noise ratio (S/N ratio), then with the Fast Median Filtering method, make it to realize quick high s/n ratio sampling, can finish quick multi channel micro-spectrophotometry testing process with the absorption light intensity (or transmitted light intensity) of each wavelength correspondence of analytical calculation computed in software and with form or the output of curve image at last.Can realize following detection index by above testing process: when (1) was 0.2mm to 2.5mm at the detection diaphragm diameter, the test sample site diameter was 1 μ m～125 μ m; (2) spectral response range reaches 300nm-1100nm; (3) wavelength resolution reaches 0.2nm; (4) finishing the full spectrum of single (300nm-800nm) detection time is 1ms.And can respectively general absorption spectrum and fluorescence spectrum be detected with the conversion of lighting source.This shows, adopt the present invention to carry out micrometering fast and accurately microscopic sample.
Embodiment 2:
The microscope of the quick multi channel micro-spectrophotometry pick-up unit of the present invention also can adopt the upright microscope of E600 type of Nikon, optical splitter adopts the reflection grating of 1500 lines/mm, the multiple tracks spectroscopic detector adopts Reticon S series line array CCD, and this line array CCD also has the synchronous data collection card simultaneously.Computing machine can adopt the compatible of band 6X86CPU and 64M internal memory and 1G hard disk, and other assembly and mutual installation and annexation are with embodiment 1; The work of this sampling device and effectiveness indicator of being obtained etc. are also similar to embodiment.
Claims (3)
1, a kind of quick multi channel micro-spectrophotometry detection method is characterized in that comprising following operation steps: (1) microscope will afterwards be composed on the multiple tracks spectroscopic detector by the optical splitter exhibition to optical splitter from the optical convergence that sample transmits; (2) each sensitization pixel of multiple tracks spectroscopic detector detects the light intensity of each wavelength; (3) the synchronous data collection card outputs to computing machine chronologically with the photoelectron signal of each sensitization pixel of multiple tracks spectroscopic detector; (4) computing machine deals with the absorption spectrum that the back obtains each wavelength to photoelectron signal.
2, quick multi channel micro-spectrophotometry detection method according to claim 1, it is characterized in that: described computing machine deals with photoelectron signal and is meant that the method for employing sample averaging earlier reduces the noisy at random of CCD multichannel detector, improve its signal to noise ratio (S/N ratio), then with the Fast Median Filtering method, make it to realize quick high s/n ratio sampling, at last with the absorption light intensity (or transmitted light intensity) of each wavelength correspondence of analytical calculation computed in software and with form or the output of curve image.
3, a kind of device of quick multi channel micro-spectrophotometry detection method, it is characterized in that: form by microscope, optical splitter, multiple tracks spectroscopic detector, synchronous data collection card, computing machine, the transmitted light of microscope detection sample is received by the multiple tracks spectroscopic detector after optical splitter exhibition spectrum, the output terminal of multiple tracks spectroscopic detector and synchronous data collection card are electrically connected, and the output terminal and the computing machine of synchronous data collection card are electrically connected.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00117473 CN1289918A (en) | 2000-10-08 | 2000-10-08 | Method and device for multi-channel spectrophotometric detection of rapid microscope |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 00117473 CN1289918A (en) | 2000-10-08 | 2000-10-08 | Method and device for multi-channel spectrophotometric detection of rapid microscope |
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| CN1289918A true CN1289918A (en) | 2001-04-04 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1837782B (en) * | 2005-03-24 | 2010-09-29 | 株式会社拓普康 | Optical image measuring apparatus |
| CN102235909A (en) * | 2010-05-07 | 2011-11-09 | 承奕科技股份有限公司 | Micro-spectroscopic device and micro-spectroscopic image capturing device |
| CN102519909A (en) * | 2011-11-14 | 2012-06-27 | 重庆大学 | Air-space low-interference phase microscope based on liquid crystal tunable filter |
| CN106289514A (en) * | 2016-07-25 | 2017-01-04 | 四川新健康成生物股份有限公司 | Multi-wavelength beam light-intensity test method, photoelectric switching circuit and biochemistry analyzer |
| WO2017193759A1 (en) * | 2016-05-11 | 2017-11-16 | 中南民族大学 | Bio-photon spectrum detection system and detection method |
| CN110489259A (en) * | 2019-07-29 | 2019-11-22 | 深圳中电长城信息安全***有限公司 | A kind of memory failure detection method and equipment |
-
2000
- 2000-10-08 CN CN 00117473 patent/CN1289918A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1837782B (en) * | 2005-03-24 | 2010-09-29 | 株式会社拓普康 | Optical image measuring apparatus |
| CN102235909A (en) * | 2010-05-07 | 2011-11-09 | 承奕科技股份有限公司 | Micro-spectroscopic device and micro-spectroscopic image capturing device |
| CN102519909A (en) * | 2011-11-14 | 2012-06-27 | 重庆大学 | Air-space low-interference phase microscope based on liquid crystal tunable filter |
| WO2017193759A1 (en) * | 2016-05-11 | 2017-11-16 | 中南民族大学 | Bio-photon spectrum detection system and detection method |
| CN106289514A (en) * | 2016-07-25 | 2017-01-04 | 四川新健康成生物股份有限公司 | Multi-wavelength beam light-intensity test method, photoelectric switching circuit and biochemistry analyzer |
| CN110489259A (en) * | 2019-07-29 | 2019-11-22 | 深圳中电长城信息安全***有限公司 | A kind of memory failure detection method and equipment |
| CN110489259B (en) * | 2019-07-29 | 2023-03-24 | 深圳中电长城信息安全***有限公司 | Memory fault detection method and equipment |
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