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Related Concept Videos

Fixation and Sectioning01:03

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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: Nov 5, 2025

Protocol for HER2 FISH Using a Non-cross-linking, Formalin-free Tissue Fixative to Combine Advantages of Cryo-preservation and Formalin Fixation
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Do Tissues Fixed in a Non-crosslinking Fixative Require a Dedicated Formalin-free Processor?

Sonia G Frasquilho1, Ignacio Sanchez1, Changyoung Yoo2

  • 1Integrated Biobank of Luxembourg, Dudelange, Luxembourg.

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
|May 19, 2021
PubMed
Summary
This summary is machine-generated.

Formalin contamination in tissue processors significantly degrades RNA yield and integrity. Alcohol-fixed tissue processed in contaminated systems shows reduced RNA quality, impacting downstream molecular analyses.

Keywords:
IlluminaPFPEScreenTapeXylenedegradationdegradefixativeformaldehydeintegrityparaffin

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Area of Science:

  • Histopathology
  • Molecular Pathology
  • Biotechnology

Background:

  • Tissue processing is crucial for diagnostic pathology.
  • Formalin fixation is a standard but can lead to cross-linking artifacts.
  • Emerging fixation methods like alcohol-based fixatives offer alternatives.

Purpose of the Study:

  • To investigate the impact of formalin contamination in tissue processors on alcohol-fixed tissue.
  • To assess the effects on nucleic acid quality (RNA and DNA), histomorphology, and immunohistochemistry.
  • To evaluate potential mitigation strategies for formalin contamination.

Main Methods:

  • Alcohol-fixed biospecimens were processed in formalin-contaminated and formalin-free tissue processors.
  • Histomorphology and immunohistochemistry were analyzed (H&E, MLH-1, Ki-67, CK-7).
  • RNA and DNA were extracted and their quality/integrity assessed (RIN, RT-PCR, qPCR, multiplex PCR).

Main Results:

  • Formalin contamination significantly reduced RNA yield (88% lower) and purity, with decreased RNA integrity number (RIN) from 5.0 to 3.8.
  • RNA amplicon length was shorter, and qPCR Cq values were higher in contaminated samples.
  • DNA integrity, histomorphology, and immunohistochemistry showed minimal to no differences between contaminated and uncontaminated groups.

Conclusions:

  • Formalin contamination in tissue processors critically impairs RNA yield and integrity.
  • While DNA and morphology are less affected, RNA quality is severely compromised.
  • Strategies to prevent or remediate formalin contamination are essential for reliable molecular diagnostics.