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Review
. 2014 Nov;70(1):12-9.
doi: 10.1016/j.ymeth.2014.01.022. Epub 2014 Feb 21.

Tissue fixation and the effect of molecular fixatives on downstream staining procedures

William J Howat  1 , Beverley A Wilson  2
Affiliations
Review

Tissue fixation and the effect of molecular fixatives on downstream staining procedures

William J Howat et al. Methods. 2014 Nov.

Abstract

It is impossible to underplay the importance of fixation in histopathology. Whether the scientist is interested in the extraction of information on lipids, proteins, RNA or DNA, fixation is critical to this extraction. This review aims to give a brief overview of the current "state of play" in fixation and focus on the effect fixation, and particularly the effect of the newer brand of "molecular fixatives" have on morphology, histochemistry, immunohistochemistry and RNA/DNA analysis. A methodology incorporating the creation of a fixation tissue microarray for the study of the effect of fixation on histochemistry is detailed.

Keywords: Histochemistry; Molecular fixatives; Tissue fixation; Tissue microarray.

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Figures

Fig. 1
Fig. 1
Graph demonstrating the differences in thickness of the oesophageal wall between each fixative. Mean and SE. Significance applied at <0.05.
Fig. 2
Fig. 2
Masson’s Trichrome staining of murine oesophagus following fixation in a variety of fixatives. Representative images of oesophageal wall, sub-mucosa and surrounding muscle are demonstrated. (A) Carnoy’s, (B) Methacarn, (C) NBF, (D) PFA, (E) Glutaraldehyde, (F) Formol Saline, (G) Zinc Formalin.
Fig. 3
Fig. 3
Graph demonstrating the analysis of percentage positivity applied to the sub-mucosal Masson’s Trichrome stained oesophagus. Each pixel is split into three “bins” of Strong, Medium and Weak positive and calculated as a percentage compared to overall pixel number.
See this image and copyright information in PMC

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