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

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...

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Published in:Histochemistry & Cell Biology Vol. 149; no. 3; pp. 209 - 219
Main Authors: Laforest, Sofia, Pelletier, Mélissa, Michaud, Andréanne, Daris, Marleen, Descamps, Justine, Soulet, Denis, Jensen, Michael D., Tchernof, André
Format: Journal Article
Published: Springer Nature Mar2018
Online Access:View this record in EBSCOhost
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      pub: Springer Nature
      place: New York, New York
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        10.1007/s00418-018-1635-3
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        atl: Histomorphometric analyses of human adipose tissues using intact, flash-frozen samples.
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          Laforest, Sofia
          Pelletier, Mélissa
          Michaud, Andréanne
          Daris, Marleen
          Descamps, Justine
          Soulet, Denis
          Jensen, Michael D.
          Tchernof, André
        affil: Endocrinology and Nephrology, CHU de Quebec-Laval University, 2705 Laurier Blvd. (R-4779), G1V 4G2, Quebec, QC, Canada
      sug:
      ab: 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 (<italic>p</italic> > 0.5). There was no difference in the distribution frequencies of adipocyte sizes in fresh-fixed vs. frozen-fixed tissues in both depots (<italic>p</italic> > 0.9). Mean adipocyte size from frozen-fixed samples correlated significantly and positively with adipocyte size from fresh-fixed samples (<italic>r</italic> = 0.74, <italic>p</italic> < 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.
      pubtype: Academic Journal
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    language: English
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