Histomorphometric analyses of human adipose tissues using intact, flash-frozen samples

Sofia Laforest, Mélissa Pelletier, Andréanne Michaud, Marleen Daris, Justine Descamps, Denis Soulet, Michael D. Jensen, André Tchernof

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Histomorphometric analyses of adipose tissue usually require formalin fixation of fresh samples. Our objective was to determine if intact, flash-frozen whole adipose tissue samples stored at − 80 °C could be used for measurements developed for fresh-fixed adipose tissues. Portions of adipose tissue samples were either formalin-fixed immediately upon sampling or flash-frozen and stored at − 80 °C and then formalin-fixed during the thawing process. Mean adipocyte diameter was measured. Immunohistochemistry was performed on additional samples to identify macrophage subtypes (M1, CD14 + and M2, CD206 +) and total (CD68 +) number. All slides were counterstained using haematoxylin and eosin (H&E). Visual inspection of H&E-stained adipose tissue slides performed in a blinded fashion showed little or no sign of cell breakage in 74% of frozen-fixed samples and in 68% of fresh-fixed samples (p > 0.5). There was no difference in the distribution frequencies of adipocyte sizes in fresh-fixed vs. frozen-fixed tissues in both depots (p > 0.9). Mean adipocyte size from frozen-fixed samples correlated significantly and positively with adipocyte size from fresh-fixed samples (r = 0.74, p < 0.0001, for both depots). The quality of staining/immunostaining and appearance of tissue architecture were comparable in fresh-fixed vs. frozen-fixed samples. In conclusion, intact flash-frozen adipose tissue samples stored at − 80 °C can be used to perform techniques conventionally applied to fresh-fixed samples. This approach allows for retrospective studies with frozen human adipose tissue samples.

Original languageEnglish (US)
Pages (from-to)209-218
Number of pages10
JournalHistochemistry and Cell Biology
Volume149
Issue number3
DOIs
StatePublished - Mar 1 2018

Keywords

  • Adipocyte hypertrophy
  • Frozen tissue
  • Image analysis
  • Immunohistochemistry
  • Macrophages

ASJC Scopus subject areas

  • Histology
  • Molecular Biology
  • Medical Laboratory Technology
  • Cell Biology

Fingerprint Dive into the research topics of 'Histomorphometric analyses of human adipose tissues using intact, flash-frozen samples'. Together they form a unique fingerprint.

  • Cite this

    Laforest, S., Pelletier, M., Michaud, A., Daris, M., Descamps, J., Soulet, D., Jensen, M. D., & Tchernof, A. (2018). Histomorphometric analyses of human adipose tissues using intact, flash-frozen samples. Histochemistry and Cell Biology, 149(3), 209-218. https://doi.org/10.1007/s00418-018-1635-3