A Super-resolution Imaging Method Based on Fluorescence Resonance Energy Transfer

A fluorescence resonance energy, super-resolution imaging technology, applied in the field of super-resolution imaging, can solve problems such as high cost and complex imaging software

Active Publication Date: 2015-11-25
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Therefore, the technical problem to be solved by the present invention is to provide a method based on fluorescence resonance energy transfer (FRET) for the defects that the existing super-resolution imaging technology requires a large amount of modification to the fluorescence microscope, the cost is very expensive, and the imaging software is also very complicated. ) super-resolution imaging method

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A Super-resolution Imaging Method Based on Fluorescence Resonance Energy Transfer
  • A Super-resolution Imaging Method Based on Fluorescence Resonance Energy Transfer
  • A Super-resolution Imaging Method Based on Fluorescence Resonance Energy Transfer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Super-resolution imaging of Cy3-Cy5 probe-labeled Hela cell microtubules

[0042] Construction of Cy3-Cy5 fluorescent probes with high FRET efficiency: Fluorescent nanospheres were prepared by inverse microemulsion method.

[0043] Experimental materials: sodium silicate solution (water glass) (the water glass modulus is 3.1, Baume degree is 40), hydrochloric acid (concentration is 1mol / L), sodium lauryl sulfate SDS, toluene, n-amyl alcohol , deionized water (W), fluorescent dye Cy3-Cy5.

[0044] The specific steps of this technique include:

[0045] Osmotic reaction: take 0.1g surfactant SDS (S), 0.2g fluorescent dye (donor Cy3 and acceptor Cy5, donor acceptor molar ratio 1:2), 0.2ml toluene and 5ml water glass solution prepared in advance Put it into a beaker, and mix it uniformly by ultrasonic (50W ultrasonic power, 1 minute mixing time) to obtain a colostrum, add n-amyl alcohol dropwise until the system is suddenly transparent, and obtain microemulsion A...

Embodiment 2

[0055] Example 2 Super-resolution imaging of Hela cell microtubules labeled with Atto488-Atto540 probe

[0056] Construction of Atto488-Atto540 Fluorescent Probe with High FRET Efficiency

[0057] Experimental materials: sodium silicate solution (water glass) (the water glass modulus is 3.2, Baume degree is 40), hydrochloric acid (concentration is 1mol / L), sodium lauryl sulfate SDS, toluene, n-amyl alcohol , deionized water (W), fluorescent dyes Atto488-Atto540.

[0058] Take 0.1g of surfactant SDS (S), 0.2g of fluorescent dye (donor Atto488 and acceptor Atto540, donor-acceptor molar ratio 1:2), 0.2ml of toluene and 5ml of water glass solution prepared in advance into a beaker In the process, ultrasonically mix evenly (ultrasonic power 50W, mixing time 1 minute) to obtain a primary emulsion, add n-amyl alcohol dropwise until the system is suddenly transparent, and obtain microemulsion A.

[0059] Polymerization reaction: replace the water glass solution with 1mol / L HCl solut...

Embodiment 3

[0067] Example 3 Super-resolution imaging of Hela cell microtubules labeled with Atto550-Atto647N probe

[0068] Construct Atto550-Atto647N fluorescent probe with high FRET efficiency:

[0069] Experimental materials: sodium silicate solution (water glass) (the water glass modulus is 3.3, Baume degree is 40), hydrochloric acid (concentration is 1mol / L), sodium lauryl sulfate SDS, toluene, n-pentanol , deionized water (W), fluorescent dye Atto550-Atto647N.

[0070] Osmotic reaction: take 0.1g surfactant SDS (S), 0.2g fluorescent dye (donor Atto550 and acceptor Atto647N, donor acceptor molar ratio 1:2), 0.2ml toluene and 5ml water glass solution prepared in advance Put it into a beaker, and mix it uniformly by ultrasonic (50W ultrasonic power, 1 minute mixing time) to obtain a colostrum, add n-amyl alcohol dropwise until the system is suddenly transparent, and obtain microemulsion A.

[0071] Polymerization reaction: replace the water glass solution with 1mol / L HCl solution, a...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
modulusaaaaaaaaaa
fluorescenceaaaaaaaaaa
fluorescenceaaaaaaaaaa
Login to view more

Abstract

A super resolution imaging method based on fluorescence resonance energy transfer, said super resolution imaging method comprises following steps: 1) marking a to-be-detected sample using high FRET effect fluorescent probe with FRET molecule pair which contains a first fluorescent group (donor) and a second fluorescent group (receptor), and the fluorescence resonance energy transfer (FRET) is transferred to the second fluorescent group from the first fluorescent group; 2) performing laser scanning con-focal microscope imaging using an excitation light threshold whose excitation light intensity enables fluorescence resonance energy thansfer of the FRET molecule pair of step 1). Said super-resolution technique based on saturated fluorescence resonance energy transfer realizes super-resolution imaging of biological sample on a genneral laser con-focal microscope, and the method can improve the resolution.

Description

technical field [0001] The invention belongs to the technical field of super-resolution imaging, in particular to a super-resolution imaging method based on fluorescence resonance energy transfer (FRET, Fluorescence Resonance Energy Transfer). Background technique [0002] Many important research achievements in modern life science rely on the progress of microscopic imaging technology. However, it has been believed for a long time that the limit resolution that optical microscopy can achieve is about 200 nanometers, which is half the wavelength of light. Until the early 1990s, with the emergence of a series of new technologies in the field of optics, the optical diffraction limit was broken and the resolution of optical microscopes was increased to tens of nanometers. [0003] The basis of super-resolution imaging technology mainly relies on various nonlinear optical effects. In 1991, a research group at Cornell University realized the multiphoton fluorescence imaging tech...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6458G01N21/6428G01N2021/6441G02B21/0076
Inventor 樊春海黄庆程亚陈建芳邓素辉梁乐
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap