Multicolor Immunofluorescence Staining of Paraffin-Embedded Human Bone Marrow Sections

Methods Mol Biol. 2021:2308:119-126. doi: 10.1007/978-1-0716-1425-9_10.

Abstract

Immunofluorescence is an indispensable method for the identification, localization and study of the expression of target antigens in formalin-fixed, paraffin-embedded (FFPE) tissue sections of human bone marrow. However, the procedure shows technical limitations because of the chemical and physical treatments required for sample processing before imaging. Here we describe a revisited protocol to obtain high-resolution images of human bone marrow trephine biopsies, improving the antigen-antibody recognition and preserving the morphology and the architecture of the bone marrow microenvironment.

Keywords: Antigen retrieval; FFPE; Fixative; Immunofluorescence; Microwave heating.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens / analysis*
  • Biomarkers / analysis
  • Biopsy
  • Bone Marrow / immunology*
  • Cellular Microenvironment
  • Color
  • Fluorescent Antibody Technique*
  • Humans
  • Microscopy, Fluorescence*
  • Microwaves
  • Paraffin Embedding*
  • Tissue Fixation

Substances

  • Antigens
  • Biomarkers