A Refractive Index Matching Method Adapted to Microfluidic-Light Sheet Imaging and Its Application

A refractive index matching, microfluidic chip technology, applied in chemical instruments and methods, laboratory containers, material analysis by optical means, etc. problems, to achieve real-time imaging compatibility, improve imaging quality, and improve flexibility

Active Publication Date: 2021-02-12
HUAZHONG UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The invention provides a refractive index matching method adapted to microfluidic-light sheet imaging and its application, which is used to solve the problem of using a salt solution or an organic solvent to adjust the existing refractive index matching method adapted to microfluidic-light sheet imaging. The refractive index of the imaging environment causes damage to biological samples and the narrow range of refractive index adjustment results in poor imaging quality

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  • A Refractive Index Matching Method Adapted to Microfluidic-Light Sheet Imaging and Its Application
  • A Refractive Index Matching Method Adapted to Microfluidic-Light Sheet Imaging and Its Application
  • A Refractive Index Matching Method Adapted to Microfluidic-Light Sheet Imaging and Its Application

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Embodiment 1

[0047] A refractive index matching method adapted to microfluidic-light sheet imaging, comprising:

[0048] Using iodixanol, adjust the imaging environment of biological sample light sheet imaging, so that the refractive index between adjacent media in the imaging environment matches;

[0049] Wherein, the adjacent media are the microfluidic chip used to fix the biological sample and the imaging liquid containing the biological sample, or the adjacent media are the water-in-oil droplets formed by the microfluidic chip and containing the biological sample. oil and water on top.

[0050] It should be noted that light sheet imaging also has the advantages of high throughput, high resolution, etc., which makes it possible to observe three-dimensional and dynamic panoramas of organisms. Light sheet imaging technology has been widely concerned and applied in the field of biological imaging. Microfluidic technology uses a microfluidic chip as a working unit. After using CAD softwar...

example 1

[0097] Example 1: Using CAD software design, mask making, photolithography, PDMS chip making and other operations, design and manufacture a microfluidic chip suitable for fixing nematode biological samples. The refractive index of the PDMS chip was tested with an Abbe refractometer; in nematode experiments, nematodes were often soaked in a specific PBS buffer, and the refractive index of the PBS buffer was tested with an Abbe refractometer; the difference between the refractive index of PBS buffer and the refractive index of PDMS There is a difference between them, and refractive index matching needs to be carried out; test the refractive index of the iodixanol solution diluted in different proportions with PBS buffer, and draw the refractive index-dilution ratio curve (such as figure 2 shown), find out the iodixanol dilution ratio corresponding to the PDMS refractive index; configure the PBS buffer dilution solution of the corresponding ratio of iodixanol, and test its refrac...

example 2

[0098] Example 2: Using CAD software design, mask making, photolithography, PDMS chip making and other operations to design and manufacture a microfluidic chip for producing large-scale water-in-oil droplets; The refractive index of the oil phase, the two have a refractive index difference, and the refractive index needs to be matched; the aqueous phase solution is selected according to different biological samples wrapped in the water-in-oil droplets, and the iodine diluted in different proportions with this aqueous phase solution is tested. Refractive index of dixanol solution, draw the refractive index-dilution ratio curve, find out the iodixanol dilution ratio corresponding to the oil phase refractive index; configure the corresponding ratio of iodixanol aqueous dilution solution, use Abbe refractometer Test its refractive index; if there is a deviation between the measured refractive index and the oil phase refractive index, according to the size of the two refractive indi...

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Abstract

The invention discloses a refractive index matching method adapted to microfluidic-light-sheet imaging and its application, including: using iodixanol to adjust the imaging environment of biological sample light-sheet imaging, so that the adjacent medium in the imaging environment Refractive index matching between them; wherein, the adjacent media are respectively the microfluidic chip used to fix the biological sample and the imaging fluid containing the biological sample, or the adjacent media are respectively the biological fluid contained in the microfluidic chip. Oil and water on a water-in-oil droplet of a sample. The present invention introduces iodixanol solution, and adjusts the refractive index of the imaging environment by changing the concentration of iodixanol. The adjustment range is wide, so that the refractive index of the adjacent medium in the imaging environment is matched, and the illumination light is eliminated between the oil phase and the water. The phase interface and the solid-liquid interface scattering phenomenon of PDMS-solution solve the problem of scattering and aberration in imaging. At the same time, iodixanol is biocompatible and can be used for real-time imaging, which effectively improves the imaging quality.

Description

technical field [0001] The invention belongs to the field of biomedical microscopic imaging, and more specifically relates to a refractive index matching method adapted to microfluidic-light sheet imaging and its application. Background technique [0002] Light-sheet imaging is a method in which a thin slice of excitation light on the micron scale is irradiated onto the sample to excite fluorescence, and is imaged by CCD or CMOS in the direction perpendicular to the excitation direction. Compared with the traditional confocal imaging method, it does not exist Excitation is out of focus, thereby minimizing phototoxicity. Microfluidic technology refers to the science and technology of high-throughput and systematic processing or control of nanoliter or picoliter-scale microfluidics in pipes with a scale of tens to hundreds of microns. Microfluidic technology combined with light-sheet imaging has two main applications: on the one hand, using microfluidic technology to generate...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/03B01L3/00
CPCB01L3/5027B01L2200/10G01N21/0303
Inventor 费鹏朱铤铤周楠森
Owner HUAZHONG UNIV OF SCI & TECH
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