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Two-color fluorescence localization super-resolution biological microscopy method and system

A super-resolution, two-color fluorescence technology, applied in the field of communication, can solve problems such as inconvenience and inconspicuousness, and achieve the effects of improving imaging quality, reducing background noise, and reducing photobleaching

Active Publication Date: 2015-04-15
宁波力显智能科技有限公司
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Problems solved by technology

Although this drift phenomenon generally exists in microscopic systems, since the resolution of general microscopes is less than 300 nanometers, the phenomenon caused by drift is not obvious
[0004] In summary, there are obviously inconveniences and defects in the actual use of the existing technology, so it is necessary to improve

Method used

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  • Two-color fluorescence localization super-resolution biological microscopy method and system
  • Two-color fluorescence localization super-resolution biological microscopy method and system
  • Two-color fluorescence localization super-resolution biological microscopy method and system

Examples

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Embodiment 1

[0086] The specific formulation of the imaging buffer in Example 1 is: 200mM Tris phosphate buffer, pH9.0; and contains 10% (w / v) glucose, 5U / ml pyranose oxidase, 57μg / ml catalase, 2 mM COT (dissolved in DMSO), 25 mM TCEP, 1 mM ascorbic acid and 1 mM methyl viologen.

Embodiment 2

[0087] The specific formulation of the imaging buffer in Example 2 is: 200mM Tris phosphate buffer, pH8.0; and contains 10% (w / v) glucose, 560μg / ml glucose oxidase, 57μg / ml catalase, 2mM COT (dissolved in DMSO), 25mM TCEP, 1mM ascorbic acid and 1mM methyl viologen.

[0088] Preferably, the signal generating module 20 is used to irradiate the biological sample with a predetermined activation laser to turn on the fluorescence signal, and irradiate the biological sample with a predetermined excitation laser to turn off the fluorescence signal, so as to generate a blinking fluorescence signal. The activation laser has a violet or ultraviolet wavelength. The excitation laser has the excitation wavelengths of Alexa647 and Alexa750 fluorescent molecules, or the excitation laser has the excitation wavelengths of Cy5 and Cy7 fluorescent molecules. The signal generation module 20 is used to use predetermined excitation laser power and activation laser power to make most of the fluoresc...

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Abstract

The invention provides a two-color fluorescence localization super-resolution biological microscopy method and a two-color fluorescence localization super-resolution biological microscopy system. The two-color fluorescence localization super-resolution biological microscopy method comprises the following steps: performing two-color fluorescence labeling on a biological sample by using Alexa647 and Alexa750 fluorescent molecules, or Cy5 and Cy7 fluorescent molecules, and soaking the biological sample in an imaging buffer solution; through laser radiation on the biological sample, generating a first channel flashing fluorescent signal corresponding to the Alexa647 fluorescent molecule or the Cy5 fluorescent molecule as well as a second channel flashing fluorescent signal corresponding to the Alexa750 fluorescent molecule or the Cy7 fluorescent molecule respectively; according to the first channel flashing fluorescent signal and the second channel flashing fluorescent signal, establishing a first biological structure super-resolution image and a second biological structure super-resolution image respectively; aligning the first biological structure super-resolution image and the second biological structure super-resolution image so as to establish a third biological structure super-resolution image. According to the technical scheme, a super-resolution biological microscopic imaging technique does not generate channel interference and can significantly reduce background noise, so that the imaging quality is improved.

Description

technical field [0001] The invention relates to the field of communication technology, in particular to a two-color fluorescence localization super-resolution biological microscopy method and system. Background technique [0002] Since Ernst Abbe (Ernst Abbe) put forward the theory of the resolution limit of optical imaging in the 1870s, people have been looking for various ways to break through this resolution limit. At present, through the use of modern cutting-edge technology, Zhuang Xiaowei and Eric Betzig (Eric Betzig) respectively proposed Stochastic Optical Reconstruction Microscopy (SORM) and Fluorescence Localization Microscopy in 2006. (Photoactivation Laser Microscopy, PLM), have achieved super-resolution imaging that breaks through ten times the optical resolution limit. Eric Betzig won the 2014 Nobel Prize in Chemistry for this technique. At present, fluorescence localization microscopy has been partially commercialized and has begun to be used in basic life s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6458G01N2021/6441G01N21/6428G01N2201/06113
Inventor 赵腾雷明德杜胜望
Owner 宁波力显智能科技有限公司
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