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Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Related Experiment Video

Updated: Oct 25, 2025

Autofluorescence Imaging to Evaluate Cellular Metabolism
07:36

Autofluorescence Imaging to Evaluate Cellular Metabolism

Published on: November 15, 2021

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Ultra-fast cycling for multiplexed cellular fluorescence imaging.

Jina Ko1, Juhyun Oh1, Maaz S Ahmed1

  • 1Center for Systems Biology, Massachusetts General Hospital Research Institute, Boston, MA 02114.

Angewandte Chemie (Weinheim an Der Bergstrasse, Germany)
|August 9, 2021
PubMed
Summary

This study introduces a rapid cycling method for single-cell analysis using tetrazine (Tz) / trans-cyclooctene (TCO) quenching. The novel approach enables multi-cycle immune cell profiling in under an hour, advancing diagnostic capabilities.

Keywords:
bioorthogonal chemistrycancerclick chemistrydiagnostic Wileyenergy transferfluorescencefluorescent probesquenchingtetrazinetetrazines

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Immunology

Background:

  • Current single-cell analysis methods are often slow and inefficient for diagnostics.
  • Rapid analysis of limited cell populations is crucial for modern healthcare.

Purpose of the Study:

  • To develop an ultra-fast, highly efficient cycling method for single-cell analysis.
  • To leverage novel linker chemistry for accelerated cellular profiling.

Main Methods:

  • Utilized tetrazine (Tz) / trans-cyclooctene (TCO) mediated quenching for single-cell analysis.
  • Implemented a multi-cycle staining protocol.
  • Focused on immune cell profiling.

Main Results:

  • Achieved quenching reaction rates over 3 orders of magnitude faster than predicted (t1/2 < 1 sec).
  • Enabled multi-cycle immune cell profiling within one hour.
  • Demonstrated the potential for rapid cellular analyses.

Conclusions:

  • The developed ultra-fast cycling method significantly enhances the speed and efficiency of single-cell analysis.
  • Accelerated kinetics using Tz/TCO quenching open new diagnostic possibilities.
  • This technique is valuable for rapid immune cell profiling and other diagnostic applications.