Abstract

Within SPIDIA, an EC FP7 project aimed to improve pre analytic procedures, the PAXgene Tissue System (PAXgene), was designed to improve tissue quality for parallel molecular and morphological analysis. Within the SPIDIA project promising results were found in both genomic and proteomic experiments with PAXgene-fixed and paraffin embedded tissue derived biomolecules. But, for this technology to be accepted for use in both clinical and basic research, it is essential that its adequacy for preserving morphology and antigenicity is validated relative to formalin fixation. It is our aim to assess the suitability of PAXgene tissue fixation for (immuno)histological methods. Normal human tissue specimens (n = 70) were collected and divided into equal parts for fixation either with formalin or PAXgene. Sections of the obtained paraffin-embedded tissue were cut and stained. Morphological aspects of PAXgene-fixed tissue were described and also scored relative to formalin-fixed tissue. Performance of PAXgene-fixed tissue in immunohistochemical and in situ hybridization assays was also assessed relative to the corresponding formalin-fixed tissues. Morphology of PAXgene-fixed paraffin embedded tissue was well preserved and deemed adequate for diagnostics in most cases. Some antigens in PAXgene-fixed and paraffin embedded sections were detectable without the need for antigen retrieval, while others were detected using standard, formalin fixation based, immunohistochemistry protocols. Comparable results were obtained with in situ hybridization and histochemical stains. Basically all assessed histological techniques were found to be applicable to PAXgene-fixed and paraffin embedded tissue. In general results obtained with PAXgene-fixed tissue are comparable to those of formalin-fixed tissue. Compromises made in morphology can be called minor compared to the advantages in the molecular pathology possibilities.

Highlights

  • Formalin has been the fixative of choice for many decades, and as a result, pathology departments have collected vast archives of formalin-fixed and paraffin embedded (FFPE) samples

  • The same evaluation was noted for thyroid and adrenal gland tissue in which the contrast between FFPE and PAXgene-fixed and paraffin embedded (PFPE) is somewhat stronger in the PAXgene-fixed tissues

  • Only minor variations are observed with slightly less contrast in the lung tissues after PAXgene fixation compared to formalin fixation

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Summary

Introduction

Formalin has been the fixative of choice for many decades, and as a result, pathology departments have collected vast archives of formalin-fixed and paraffin embedded (FFPE) samples. These archives of well-defined and documented tissue samples are frequently used in medical research [1]. While (mi)RNA, DNA and proteins can be isolated from FFPE samples [3,4,5,6,7,8], assay reproducibility; diagnostic value may be limited This is due to the poor quality of the derivates caused by varying fixation times which results in high variation of the number of cross-links in any individual sample [9]

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