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Updated: Mar 2, 2026

Optimization, Design and Avoiding Pitfalls in Manual Multiplex Fluorescent Immunohistochemistry
Published on: July 26, 2019
Optimization of Immunostaining for Prospective Image Analysis
Scott M Lawrence1, Yelena G Golubeva2
1Pharmacodynamics Assay Section (PADIS), Laboratory of Human Toxicology and Pharmacology, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD, USA. Scott.Lawrence@fnlcr.nih.gov.
Abstract:
Biomarker discovery is a crucial part of the fast developing field of personalized medicine. Antibody-based techniques including immunostaining of tissue samples are widely used for biomarker evaluation in preclinical and clinical studies. When used in conjunction with robust image analysis methods, it provides a powerful means to assess biomarker modulation, toxicity, and patient response to targeted agents. Here, we describe the optimization of immunofluorescent (IF) staining protocols and a sample IF multiplex protocol suitable for colocalization image analysis.

