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Related Concept Videos

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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 developed.
Cryo-electron Microscopy01:28

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

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Related Experiment Video

Updated: May 9, 2026

High-Throughput, Multi-Image Cryohistology of Mineralized Tissues
10:18

High-Throughput, Multi-Image Cryohistology of Mineralized Tissues

Published on: September 14, 2016

High-resolution multiphoton cryomicroscopy.

Karsten König1, Aisada Uchugonova2, Hans Georg Breunig1

  • 1JenLab GmbH, Schillerstrasse 1, 07745 Jena, Germany; Department of Biophotonics and Laser Technology, Saarland University, Campus A5.1, 66123 Saarbrücken, Germany.

Methods (San Diego, Calif.)
|July 23, 2013
PubMed
Summary

This study used multiphoton cryomicroscopy to observe cellular autofluorescence changes during freezing. Researchers found that cooling alters cell fluorescence and morphology, enabling label-free optical biopsies.

Keywords:
BiobankCryomicroscopeCryopreservationFLIMFreezingMultiphoton imagingTwo-photon

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Area of Science:

  • Biophysics
  • Cell Biology
  • Optical Microscopy

Background:

  • Cellular responses to freezing are critical for cryopreservation.
  • Understanding autofluorescence changes provides insights into cellular state.
  • Label-free imaging methods are desirable for non-destructive analysis.

Purpose of the Study:

  • To investigate cellular autofluorescence and morphology during freezing and thawing.
  • To evaluate the use of multiphoton cryomicroscopy for studying cryobiological processes.
  • To demonstrate label-free optical biopsy capabilities.

Main Methods:

  • Utilized an ultracompact high-resolution multiphoton cryomicroscope.
  • Employed a femtosecond near-infrared fiber laser for imaging.
  • Studied cellular autofluorescence and morphology during controlled cooling and thawing, with and without DMSO.

Main Results:

  • Intracellular fluorescence intensity increased upon cooling.
  • Morphological modifications were observed below -10 °C, influenced by DMSO and cooling rate.
  • Fluorescence lifetime imaging showed increased mean lifetime with decreasing temperature.
  • Non-destructive, label-free optical biopsies were achieved with low laser power.

Conclusions:

  • Multiphoton cryomicroscopy effectively visualizes cellular changes during cryopreservation.
  • Autofluorescence and its lifetime are sensitive indicators of cellular state during freezing.
  • The technique offers potential for label-free, non-destructive assessment of biomaterials in ice.