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Scattered light and fluorescent light bimodal flow imaging system

A technology of scattering fluorescence and imaging system, which is applied in the field of optical instruments, can solve problems such as identification difficulties, cell size counting errors, insufficient spectral imaging channels, etc., and achieve the effect of expanding the scope of application and compact structure

Active Publication Date: 2019-08-13
深圳市趣方科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, light sheet fluorescence flow imaging technology still has many limitations in solving the problem of plankton observation
First of all, at present, this technology can only rely on the autofluorescence of certain pigments in plankton as an imaging contrast mechanism, so it is difficult to achieve accurate imaging measurements of targets or parts that cannot emit autofluorescence, which limits its application to most zooplankton and zooplankton. Prospects for Bacteria
Secondly, many phytoplankton, especially micro and small eukaryotic cells above 20 μm, usually have transparent structures such as cell walls and flagella without fluorescent pigments. These morphological structures are often unique characteristics of specific phytoplankton species; these characteristics cannot be observed Difficulty not only for subsequent identification but also errors in cell size and / or count measurements
Finally, the development of various molecular techniques can not only fluorescently label plankton without autofluorescent pigments, but also greatly increase its detection specificity; however, the existing light-sheet fluorescence imaging flow cytometry has a single excitation wavelength and spectral imaging Insufficient channels to meet the demand for multispectral imaging detection of more different fluorescent labels in a wider spectral range

Method used

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  • Scattered light and fluorescent light bimodal flow imaging system
  • Scattered light and fluorescent light bimodal flow imaging system
  • Scattered light and fluorescent light bimodal flow imaging system

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0047] Example 1: Detection of marine phytoplankton cells

[0048] As attached to the manual figure 2 As shown, the multicolor orthogonal lighting unit A specifically includes two sets of lasers 11, 12 and beam shaping modules 21, 22 that cooperate with each other. The center wavelengths of the laser 11 and the laser 12 are 445nm (blue light b) and 532nm (green light g) respectively. ), corresponding to the excitation peaks of chlorophyll a and phycoerythrin in marine phytoplankton cells, respectively. Taking the excitation light path of blue light b as an example, after passing through the beam shaping module 21, the Gaussian beam emitted by the laser 11 is shaped and the illumination direction is adjusted to be perpendicular to the flow direction of the sample in the sample tube 3, and the illumination beam coincides with the focal plane of the imaging objective lens 4 , perpendicular to the flow side of the injection. The phytoplankton cell particles will scatter the ill...

example 2

[0049] Example 2: For the detection of freshwater phytoplankton cells

[0050] The main difference between freshwater phytoplankton and seawater phytoplankton is that the ratio of phycoerythrin and phycocyanin in the cells is different. Many marine phytoplankton cells contain phycoerythrin, while freshwater phytoplankton cells contain most phycocyanin. Therefore, Example 2 and The main difference of Example 1 lies in the excitation and collection of phycoerythrin / phycocyanin autofluorescence.

[0051] As attached to the manual image 3 As shown, the multi-color orthogonal lighting unit A specifically includes two sets of lasers 11, 13 and beam shaping modules 21, 23 that work together. The center wavelengths of the laser 11 and the laser 13 are 445nm (blue light b) and 633nm (red light r) respectively. ), corresponding to the excitation peaks of chlorophyll a and phycocyanin in freshwater phytoplankton cells, respectively. Taking the red excitation light path as an example, ...

example 3

[0052] Example 3: For the detection of various phytoplankton cells

[0053] If two sets of systems are needed to observe and analyze seawater phytoplankton and freshwater phytoplankton, it is obviously troublesome. Therefore, the excitation and collection methods of phycoerythrin and phycocyanin autofluorescence in the above-mentioned embodiments can be combined for a comprehensive analysis of seawater and freshwater phytoplankton in one system.

[0054] As attached to the manual Figure 4 As shown, the multi-color orthogonal lighting unit A specifically includes three sets of lasers 11, 12, 13 and beam shaping modules 21, 22, 23 used in conjunction with each other. In this embodiment, three lasers are used to cover more excitation peaks. The center wavelengths are 445nm (blue light b), 532nm (green light g) and 633nm (red light r) respectively. Taking the green light excitation optical path as an example, after passing through the beam shaping unit 22, the Gaussian beam emi...

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Abstract

The invention provides a scattered light and fluorescent light bimodal flow imaging system. The system comprises a multicolor orthogonal lighting unit, a multichannel microimaging unit and a feeding tube, wherein the multicolor orthogonal lighting unit comprises an excitation light source and a beam shaping module, the multichannel microimaging unit comprises an objective lens, a multispectral image divider and multiple cameras, and the feeding tube is coaxial with the objective lens. The excitation light source can emit exciting light with two or more wavelengths, and the exciting light perpendicularly illuminates the feeding tube at the position overlapping with a focal plane of the objective lens after being shaped by the beam shaping module. The objective lens collects scattered lightand fluorescent light which are emitted by bio-particles, and the multispectral image divider projects the scattered light and the fluorescent light to the different cameras for imaging through a scattering channel and one or more fluorescent light channels according to a specific spectral passband. According to the whole system, the principle is based on a lateral scattered light and fluorescentlight emission microimaging technology of light-sheet lighting, a more accurate plankton observation result can be obtained, and the system can adapt to a wider-spectrum water plankton species range and meet the online measurement demand on non-living particulate matter.

Description

technical field [0001] The invention belongs to the optical instrument technology in the field of biology and environment, and in particular relates to a novel flow imaging system. Background technique [0002] Plankton is the basic component of aquatic ecosystems and plays an important role in the material cycle and energy flow of the entire food chain. Understanding the physiology, ecology, diversity, and processes of plankton is a fundamental requirement for research on marine resources, biodiversity, and ecosystem responses to climate change. However, although the existing plankton observation technology and observation platform have been developed to a certain extent, the detection throughput, specificity and accuracy of the identification and quantification technology are still seriously insufficient, which is the current and future related scientific research and water environment monitoring needs. One of the main challenges faced. [0003] In addition to the vastne...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N15/14
CPCG01N21/6428G01N15/14G01N2021/6439G01N2021/6491G01N2015/144Y02A90/10
Inventor 李军陆昱
Owner 深圳市趣方科技有限公司
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