Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

protein retention extension microscopy

A protein and fluorescent protein technology, applied in the field of protein retention extended microscopy, can solve the problems of customized synthesis obstacles, inability to customize reagents, and inability to image fluorophores

Active Publication Date: 2020-08-28
MASSACHUSETTS INST OF TECH
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

All chemicals required for ExM are commercially available except for the gel-anchorable marker, which requires custom synthesis and presents a barrier to researchers adopting the described method
Another disadvantage of the ExM protocol is that genetically encoded fluorophores cannot be imaged without antibody labeling
Additionally, ExM cannot maintain native proteins in gels and uses custom reagents that are not widely available

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
  • protein retention extension microscopy
  • protein retention extension microscopy
  • protein retention extension microscopy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0096] raw material solution

[0097] 4% paraformaldehyde

[0098] 4% Paraformaldehyde (from Electron Microscopy Science 16% stock)

[0099] 1x PBS

[0100] Quench solution (store at 4C for use over extended periods of time)

[0101] 1x PBS

[0102] 100mM Glycine

[0103] protein anchor solution

[0104] 1x PBS

[0105] 0.1 mg / mL 6-((acryloyl)amino)caproic acid, succinimidyl ester (acryloyl-X, SE)

[0106] Tissue Disruption Solution (Autoclaved Version)

[0107] 100mM Tris base

[0108] 1% sodium lauryl sulfate

[0109] 5% Triton X-100

[0110] Tissue Disruption Solution (Phospholipase Version)

[0111] 0.5x PBS

[0112] 0.1% Triton X-100

[0113] Phospholipase A1 (Sigma, L3295) 100U / mL

[0114] Phospholipase D (Enzo, BML-SE301-0025) 500U / mL

[0115] Antibody staining solution (stored at 4C, can be used for at least 1 month)

[0116] 1x PBS

[0117] 0.1% Triton X-100

[0118] 2% normal donkey serum

[0119] Monomer solution:

[0120]

[0121]

[012...

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

No PUM Login to View More

Abstract

The present invention provides a method called protein retention ExM (proExM), in which cross-linking molecules are used to anchor proteins, rather than markers, to a swellable gel. This proExM strategy can be used to perform nanoscale imaging of immunostained cells and tissues as well as FP-expressing samples, since even when subjected to non-specific proteolytic digestion, genetically encoded fluorescent proteins and / or conventional fluorescent proteins anchored directly to the gel Fluorescent signals from labeled secondary antibodies and streptavidin were maintained.

Description

[0001] related application [0002] This application claims the benefit of priority to US Provisional Application Serial No. 62 / 202,423, filed August 7, 2015, the contents of which are incorporated herein by reference in their entirety. [0003] Government funding [0004] This invention was made with government support under NYSCF-R-NI10 awarded by the Hertz Foundation, NYSCF, NSF, and Rehabilitation Institute of Chicago, and 1-U01-MH106011 awarded by the NIH and Cargill Fund Bioengineering Fund. The government has certain rights in this invention. Background technique [0005] Extended microscopy (ExM) can image thick preserved specimens with ~70nm lateral resolution. With ExM, the optical diffraction limit is circumvented by physically expanding the biological sample prior to imaging, thereby bringing sub-diffraction-limited structures into a size range observable by conventional diffraction-limited microscopy. ExM can image biological specimens at the voxel rate of diff...

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): G01N33/68G01N1/36
CPCG01N1/36G01N33/6803G01N2333/95G01N1/30G01N33/582G01N2001/302G01N2001/307
Inventor P·W·蒂尔伯格F·陈E·S·博伊登C-C·余
Owner MASSACHUSETTS INST OF TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com