An Introduction to Performing Immunofluorescence Staining

  • Methods Mol Biol. 2019:1897:299-311. doi: 10.1007/978-1-4939-8935-5_26.
Kyuseok Im  1  2 ,  Sergey Mareninov  1  2 ,  M Fernando Palma Diaz  1 ,  William H Yong  3  4  5
Affiliations
  • 1. Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
  • 2. Brain Tumor Translational Resource, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
  • 3. Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA. [email protected].
  • 4. Brain Tumor Translational Resource, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA. [email protected].
  • 5. Jonsson Comprehensive Cancer Center, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA. [email protected].
PMID: 30539454 DOI: 10.1007/978-1-4939-8935-5_26
Abstract

Immunofluorescence (IF) is an important immunochemical technique that allows for detection and localization of a wide variety of Antigens in different types of tissues of various cell preparations. IF allows for excellent sensitivity and amplification of signal in comparison to immunohistochemistry, employing various microscopy techniques. There are two methods available, depending on the scope of the experiment or the specific antibodies in use: direct (primary) or indirect (secondary). Here, we describe preparation of specimens preserved in different types of media and step-by-step methods for both direct and indirect immunofluorescence staining.

Keywords
Antigen retrieval; Fixation; Fluorescence; Fluorophore; Immunocytochemistry; Immunofluorescence; Immunohistochemistry.