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Laser fluorescence correlation spectrum unimolecular analyzer

A fluorescence correlation spectroscopy and single-molecule technology, which is applied in the fields of life science, analytical chemistry detection technology and optical instruments, can solve the problems of complex mode, complex adjustment process, and large loss of fluorescent signal, and achieve high sensitivity, good stability, and easy operation. simple effect

Inactive Publication Date: 2005-04-13
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This mode is complicated, and it is difficult to ensure that the photosensitive area of ​​the detector, the pinhole, and the focal point of the objective lens are strictly coaxial, so the loss of fluorescence signal is also large, and the adjustment process is very complicated

Method used

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  • Laser fluorescence correlation spectrum unimolecular analyzer
  • Laser fluorescence correlation spectrum unimolecular analyzer
  • Laser fluorescence correlation spectrum unimolecular analyzer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Fluorescein analysis:

[0023] Using a 35 μm pinhole, the sample implementation process was as described above. attached figure 2 The fluorescence correlation spectrum of fluorescein is given. figure 2 In a, there are 0.4 fluorescein molecules in the irradiated micro-area, figure 2 There are 2 fluorescein molecules in b.

Embodiment 2

[0025] Rhodamine Green Analysis:

[0026] Using a 35 μm pinhole, the sample implementation process was as described above. attached image 3 The fluorescence correlation spectrum of rhodamine green is given. image 3 In a, there are 0.3 rhodamine green molecules in the irradiated micro-area, image 3 There are 2 rhodamine green molecules in b.

Embodiment 3

[0028] Goat anti-human IgG-FITC assay:

[0029] Using a 160 μm pinhole, the sample implementation process was as described above. attached Figure 4 The fluorescence correlation spectrum of goat anti-human IgG-FITC is given. Figure 4 There are 8 goat anti-human IgG-FITC molecules in the medium-irradiated microzone.

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Abstract

The laser fluorescent correlating spectrum single molecular analyzer is used in the research of single molecule inside solution in life science, chemistry, physics and other fields. The sample solution is set on cover glass or sample pond; the laser beam, after being beam expanded, filtered in exciting filter, reflected in the dichromatic mirror and focused in the objective of microscope, irradiates the sample solution to excite the molecule under research; the fluorescence the sample solution emits is made to pass through the same objective, the dichromatic mirror, emitting filter to eliminate hybrid light, lens to focus and pinhole coupled to reach the single photon detector; and the signal the single photon detector generates is acquired with the data acquisition card and processed in the computer. The present invention has the features of simple operation, high stability, less light loss, high sensitivity, etc. and may be used in various basic researches.

Description

technical field [0001] The present invention relates to a single-molecule analyzer, in particular to a laser fluorescence correlation spectrum single-molecule analyzer based on Fluorescence Correlation Spectroscopy (FCS) technology, which is used in solutions in the fields of life science, chemistry, physics, etc. The study of a single molecule belongs to the fields of life sciences, analytical chemistry detection technology and optical instruments. Background technique [0002] Single-molecule detection has reached the limit of molecular detection, which has been pursued by people for a long time. It has important scientific significance and application prospects in the fields of life science, chemistry, and physics. This method can obtain important information that cannot be obtained by traditional methods at the single molecule level, and is especially suitable for the study of chemical and biochemical reaction kinetics, biomolecular interactions, structural and functiona...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 任吉存张普敦
Owner SHANGHAI JIAO TONG UNIV
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