Optimization of Immunostaining for Prospective Image Analysis

  • Scott M. LawrenceEmail author
  • Yelena G. Golubeva
Part of the Methods in Molecular Biology book series (MIMB, volume 1606)


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.

Key words

Antibodies Immunofluorescence Multiplex IF Image acquisition Image Analysis 



This Research was supported (in part) by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research. This project has been funded in whole or in part with federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US government. The authors would like to thank staff of Clinical Pharmacodynamic Biomarkers Program, Applied/Developmental Research Directorate, and also Donna Butcher, Brad Gouker, and Simona Florea of Pathology-Histotechnology Laboratory, Leidos Biomedical Research, Inc., and Frederick National Laboratory for Cancer Research, USA, for technical assistance.


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Pharmacodynamics Assay Section (PADIS), Laboratory of Human Toxicology and PharmacologyLeidos Biomedical Research, Inc., Frederick National Laboratory for Cancer ResearchFrederickUSA
  2. 2.MedImmuneGaithersburgUSA

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