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Related Concept Videos

Fixation and Sectioning01:03

Fixation and Sectioning

6.0K
Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
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Preparation of Samples for Electron Microscopy01:20

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To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
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Related Experiment Video

Updated: May 1, 2026

Proteomic Sample Preparation from Formalin Fixed and Paraffin Embedded Tissue
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Proteomic Sample Preparation from Formalin Fixed and Paraffin Embedded Tissue

Published on: September 2, 2013

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Formaldehyde Fixation Helps Preserve the Proteome State during Single-Cell Proteomics Sample Processing and Analysis.

Ilaria Piga1, Claire Koenig1, Maico Lechner1

  • 1Novo Nordisk Foundation Center for Protein Research, Proteomics Program, Faculty of Health and Medical Sciences, University of Copenhagen, 2200 Copenhagen, Denmark.

Journal of Proteome Research
|February 4, 2025
PubMed
Summary

Short-term formaldehyde fixation preserves cell states for mass spectrometry-based single-cell proteomics (SCP). This method enables sample shipping and prolonged cell sorting, making SCP more accessible for biological research.

Keywords:
One-Tipcell-sortingformaldehyde fixationsingle-cell proteomics

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Area of Science:

  • Proteomics
  • Cell Biology
  • Biotechnology

Background:

  • Mass spectrometry-based single-cell proteomics (SCP) is a powerful technique but limited by high costs and expertise.
  • Current SCP workflows require immediate processing, hindering sample sharing and prolonged analysis.
  • Lack of standardized methods for preserving cell proteomes during sample transfer restricts SCP accessibility.

Purpose of the Study:

  • To evaluate the efficacy of short-term formaldehyde (FA) fixation in preserving cell integrity and proteome state for SCP.
  • To determine if FA fixation impacts protein recovery and analytical depth in SCP workflows.
  • To assess the compatibility of FA-fixed samples with prolonged cell sorting and downstream SCP analysis.

Main Methods:

  • Short-term formaldehyde (FA) fixation of cells.
  • Mass spectrometry-based single-cell proteomics (SCP) analysis.
  • Comparison of FA-fixed versus unfixed samples regarding protein recovery, analytical depth, and preservation of drug-induced perturbations.
  • Evaluation of sample handling during fluorescence-activated cell sorting (FACS).

Main Results:

  • Short-term FA fixation maintained cell integrity and proteome state without significant impact on protein recovery or analytical depth.
  • FA fixation did not require harsh conditions like heating or strong detergents.
  • Drug-induced protein abundance changes were preserved in FA-fixed samples after cell sorting.
  • FA fixation proved compatible with prolonged cell sorting procedures.

Conclusions:

  • Short-term formaldehyde fixation is a viable method to stabilize cell proteomes for SCP.
  • FA fixation facilitates sample shipping and prolonged cell sorting, broadening access to SCP technology.
  • This approach can democratize SCP, accelerating discoveries in cell biology and personalized medicine.