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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...
Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

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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Processing Embryo, Eggshell, and Fungal Culture for Scanning Electron Microscopy
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Published on: August 16, 2019

Special symposium: fixation and tissue processing models.

W E Grizzle1

  • 1Department of Pathology, University of Alabama at Birmingham, Zeigler Research Building, ZRB 408, 703 South 19th Street, Birmingham, AL 35294-0007, USA. wgrizzle@uab.edu

Biotechnic & Histochemistry : Official Publication of the Biological Stain Commission
|November 6, 2009
PubMed
Summary
This summary is machine-generated.

10% neutral buffered formalin (10% NBF) is the standard fixative for tissue blocks, but its interaction with tissue processing and modern biomarker assays is poorly understood. This review highlights challenges and limitations in using 10% NBF for immunohistochemistry and other molecular analyses.

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Area of Science:

  • Histopathology
  • Biomarker Analysis
  • Tissue Processing

Background:

  • 10% neutral buffered formalin (10% NBF) is the gold standard fixative in pathology for tissue preservation and microscopic examination.
  • Pathologists are hesitant to adopt new fixatives due to established training and familiar microscopic appearances.
  • Archival tissue blocks are predominantly fixed in 10% NBF, making it crucial for retrospective molecular studies.

Purpose of the Study:

  • To review the challenges and limitations associated with using 10% NBF in combination with tissue processing for biomarker analysis.
  • To highlight the impact of fixation and processing on immunohistochemistry and other molecular assays.
  • To discuss the interaction between fixation duration, tissue processing, and biomarker detection.

Main Methods:

  • Review of existing literature on tissue fixation, processing, and biomarker analysis.
  • Discussion of the effects of 10% NBF fixation on histochemical and immunohistochemical staining.
  • Examination of the impact of short fixation times and tissue processing on molecular assays.

Main Results:

  • Limited studies exist on the interaction between 10% NBF fixation (especially short durations) and tissue processing.
  • Many antibody-antigen combinations show reduced or lost immunorecognition in tissues fixed with 10% NBF.
  • Existing models often attribute immunorecognition issues solely to fixation, neglecting the role of processing.

Conclusions:

  • The combined effects of 10% NBF fixation and tissue processing are critical for accurate biomarker analysis.
  • Further research is needed to understand and optimize protocols for 10% NBF-fixed tissues in modern molecular assays.
  • Developing models that account for both fixation and processing is essential for reliable retrospective studies.