Lasing fluorescence scanning imaging-fluorescence correlation spectrum unimolecule detecting instrument

Abstract

The invention relates to a lasing fluorescence scanning imaging-fluorescence correlation spectrum unimolecule detecting instrument. Cover glass or a sample cell is arranged on an X-Y scanning platform; a microscope objective is installed on a Z-direction locator. Expended laser is filtered by an excitation filter, and then reflected into the objective by a dichroic mirror, and finally radiated in a sample above the cover glass after being focused by the objective; the sample generates fluorescence; and the generated fluorescence is collected by the same objective, passes through the dichroic mirror, and stray light is filtered off by an emission filter, and finally the filtered fluorescence is focused by a focusing lens on a small hole which is coupled with a single photon detector; and a signal generated by the single photon detector enters a computer through a data acquisition card. In the computer, three-dimension displacement and positioning system control software control the X-Y scanning platform to do X-Y two-dimension movement, and the Z-direction locator controls the focus of the objective to do Z-direction movement, thereby forming the three-dimension scanning to the sample; and three-dimension space position information and the signal generated by the single photon detector construct three-dimension scanning images and fluorescence correlation spectrum curves in the computer.

Description

technical field [0001] The invention relates to a laser fluorescence scanning imaging-fluorescence correlation spectrum single-molecule detector, which is a device combining laser confocal scanning imaging (Laser Confocal Scanning Imaging) and fluorescence correlation spectroscopy (Fluorescence Correlation Spectroscopy, FCS) technology, It is used for fluorescence scanning imaging and single-molecule detection research in the fields of life science, chemistry, physics, etc., and belongs to the field of life science, analytical chemistry detection technology and optical instruments. Background technique [0002] Cell is the basic unit of life activities, and also the smallest unit for human beings to regulate life activities. But the cell is a very small and complex biological system. A simple cell has a diameter of only a few microns to tens of microns, but it contains thousands of molecules of different sizes. Each molecule participates in a large number of chemical react...

AI Technical Summary

Problems solved by technology

First, its scanning accuracy and recovery are low (hundreds of nanometers)

Second, optical deformation occurs when scanning the non-objective focal position (the optical configuration of the non-objective focal region no longer has a three-dimensional Gaussian distribution), resulting in in-situ fluorescence correlation spectroscopy single-molecule detection quantitative analysis after scanning imaging The accuracy is poor, and the obtained fluorescence-related spectral information has a large deviation (such as detecting changes in the size and shape of the micro-area when analyzing the focal area of ​​the non-objective lens, etc.)

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