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Related Concept Videos

Fixation and Sectioning01:03

Fixation and Sectioning

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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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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Will PAXgene substitute formalin? A morphological and molecular comparative study using a new fixative system.

Benedetta Belloni1, Chiara Lambertini, Paolo Nuciforo

  • 1Department of Dermatology, University Hospital of Zurich, Zurich, Switzerland.

Journal of Clinical Pathology
|November 6, 2012
PubMed
Summary

The PAXgene tissue system offers better RNA integrity than formalin fixation but requires re-evaluation of immunohistochemistry markers. It provides molecular analysis advantages but doesn't justify replacing formalin fixation in routine pathology labs.

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Area of Science:

  • Histopathology and Molecular Diagnostics
  • Tissue Preservation Techniques
  • Cancer Research

Background:

  • Formalin fixation and paraffin embedding (FFPE) are standard for tissue analysis but formalin cross-linking hinders molecular studies.
  • The PAXgene tissue system offers a formalin-free alternative for tissue preservation.

Purpose of the Study:

  • To compare the PAXgene tissue system with formalin fixation and fresh frozen samples for tissue analysis.
  • To evaluate morphology, immunohistochemistry, and molecular integrity (DNA/RNA) of tissues preserved with different methods.

Main Methods:

  • Melanoma biopsy samples (n=12) were divided and processed using formalin, PAXgene, or fresh frozen methods.
  • Analyses included morphology, immunohistochemistry for differentiation/proliferation markers, DNA, and RNA integrity assessments.

Main Results:

  • PAXgene samples showed well-preserved morphology and superior RNA integrity, with less DNA fragmentation compared to FFPE.
  • Immunohistochemistry staining intensity was lower for 5 out of 11 markers in PAXgene samples, but could be improved with detergent treatment.
  • DNA sequencing and mutational analysis were comparable across all fixation methods.

Conclusions:

  • PAXgene fixation preserves RNA integrity better than formalin fixation, beneficial for molecular analysis.
  • Switching to PAXgene requires re-optimization of immunohistochemical protocols.
  • While offering molecular advantages, PAXgene fixation is not recommended for routine pathology substitution of FFPE.