Comparison of Vibratome and Compresstome sectioning of fresh primate lymphoid and genital tissues for in situ MHC-tetramer and immunofluorescence staining

Hadia M. Abdelaal, Hyeon O. Kim, Reece Wagstaff, Ryoko Sawahata, Peter Southern, Pamela J Skinner

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Background: For decades, the Vibratome served as a standard laboratory resource for sectioning fresh and fixed tissues. In skilled hands, high quality and consistent fresh unfixed tissue sections can be produced using a Vibratome but the sectioning procedure is extremely time consuming. In this study, we conducted a systematic comparison between the Vibratome and a new approach to section fresh unfixed tissues using a Compresstome. We used a Vibratome and a Compresstome to cut fresh unfixed lymphoid and genital non-human primate tissues then used in situ tetramer staining to label virus-specific CD8 T cells and immunofluorescent counter-staining to label B and T cells. We compared the Vibratome and Compresstome in five different sectioning parameters: speed of cutting, chilling capability, specimen stabilization, size of section, and section/staining quality. Results: Overall, the Compresstome and Vibratome both produced high quality sections from unfixed spleen, lymph node, vagina, cervix, and uterus, and subsequent immunofluorescent staining was equivalent. The Compresstome however, offered distinct advantages; producing sections approximately 5 times faster than the Vibratome, cutting tissue sections more easily, and allowing production of larger sections. Conclusions: A Compresstome can be used to generate fresh unfixed primate lymph node, spleen, vagina, cervix and uterus sections, and is superior to a Vibratome in cutting these fresh tissues.

Original languageEnglish (US)
Article number2
JournalBiological Procedures Online
Volume17
Issue number1
DOIs
StatePublished - Jan 7 2015

Bibliographical note

Funding Information:
We thank the NIH Tetramer Facility for providing MHC monomers used in this study. Public Health Services Grants 5 R01 AI090732, R01AI096966, and P01 AI095985 supported this work.

Publisher Copyright:
© 2015 Abdelaal et al.; licensee BioMed Central.

Keywords

  • Cervix
  • Compresstome
  • Immunohistochemistry
  • In situ tetramer staining
  • Lymph node
  • Spleen
  • Unfixed fresh tissue sectioning
  • Uterus
  • Vagina
  • Vibratome

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