Unimolecular tracing method in low viscosity solution

A single-molecule, low-viscosity technology, used in individual particle analysis, material excitation analysis, particle and sedimentation analysis, etc., it can solve the problems of complex methods, high cost, and inability to track multiple single molecules, achieving simple operation and low cost. low cost effect

Inactive Publication Date: 2010-12-29
HUNAN UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0008] Although these instruments can track single molecules to a certain extent, they are costly, complicated, and have unsatisfactory signal-to-noise ratios. They cannot track multiple single molecules reacting at the same time, which will interfere with the reaction process.

Method used

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  • Unimolecular tracing method in low viscosity solution
  • Unimolecular tracing method in low viscosity solution
  • Unimolecular tracing method in low viscosity solution

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Embodiment Construction

[0045] Add 5 microliters of Alexa488-labeled bovine serum albumin (BSA) samples onto the surface of the dehydrated agarose gel, cover with a cover slip, and send to a fluorescent microscope for observation. Measure the sedimentation coefficient distribution of the 600nM sample, the solid line is the reduced state, and the dotted line is the non-reduced state (see Figure 3); the non-denatured polyacrylamide electrophoresis image of the 6000nM sample (see Figure 4); Photobleaching process and fluorescence lifetime distribution (see Figures 5 and 6). The results of mass spectrometry showed that Alexa488 was labeled in Lys 211 (40% coverage). The diffusion of a single protein molecule at a concentration of 80pM is tracked by imaging, and the diffusion trajectory of a single molecule is obtained (see Figure 7). Stay within 2.6±2.6 seconds. Investigated the BSA diffusion coefficient under this experimental condition (see Figure 9, the slope is), D=0.73±0.71 μm 2 / s, which is 80 ...

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Abstract

The invention relates to single molecule tracing method in low viscosity solution. It uses dye marked molecule as tracing object, dehydrated agarose gel to decorate slide, processes fluorescence imaging for the single molecule in the solution between the cover glass and agarose gel. It includes the following steps: forming agarose gel film on the slide; covering the cover glass; sending to obverse by the microscope to do single molecule fluorescence imaging. The method of invention can realize dynamical real time monitoring, has low cost, simple and fast operation.

Description

Technical field: [0001] The invention mainly relates to a single-molecule tracing method in a low-viscosity solution. Background technique: [0002] So far, what traditional analytical chemistry has measured is actually the ensemble average of a large number of molecules, and the statistical results of this ensemble can no longer meet the analysis requirements for complex and non-homogeneous biological systems. In recent years, an analytical technique that targets single molecules has emerged. Detection technology at the single-molecule level has become a powerful supplement to traditional ensemble research methods, based on individual molecules, to study macroscopic systems. Compared with the ensemble method which only provides the average properties of the system, single-molecule detection technology can detect the distribution and time trajectory information of physical quantities, and can study the fluctuation of the system in equilibrium and the reaction pathway in non...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N15/10G01N21/64
Inventor 盖宏伟李群
Owner HUNAN UNIV
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