Please use this identifier to cite or link to this item: https://doi.org/10.3791/58711
Title: Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
Authors: Hong, Guo
Fan, Shuangyi 
Phyu, The 
Maheshwari, Priyanka 
Hoppe, Michal Marek
Hoang, Mai Phuong 
de Mel, Sanjay
Poon, Michelle
Ng, Siok-Bian 
Jeyasekharan, Anand D 
Keywords: Science & Technology
Multidisciplinary Sciences
Science & Technology - Other Topics
Immunology and Infection
Issue 143
Multiplexed immunofluorescence
multispectral imaging
quantitative analysis
biomarkers
lymphoma
digital pathology
TYRAMIDE SIGNAL AMPLIFICATION
Issue Date: 1-Jan-2019
Publisher: JOURNAL OF VISUALIZED EXPERIMENTS
Citation: Hong, Guo, Fan, Shuangyi, Phyu, The, Maheshwari, Priyanka, Hoppe, Michal Marek, Hoang, Mai Phuong, de Mel, Sanjay, Poon, Michelle, Ng, Siok-Bian, Jeyasekharan, Anand D (2019-01-01). Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS 2019 (143). ScholarBank@NUS Repository. https://doi.org/10.3791/58711
Abstract: Immunohistochemical (IHC) methods for the in-situ analysis of protein expression by light microscopy are a powerful tool for both research and diagnostic purposes. However, the visualization and quantification of multiple antigens in a single tissue section using conventional chromogenic IHC is challenging. Multiplexed imaging is especially relevant in lymphoma research and diagnostics, where markers have to be interpreted in the context of a complex tumor microenvironment. Here we describe a protocol for multiplexed fluorescent IHC staining to enable the quantitative assessment of multiple targets in specific cell types of interest in lymphoma.The method covers aspects of antibody validation, antibody optimization, the multiplex optimization with markers of lymphoma subtypes, the staining of tissue microarray (TMA) slides, and the scanning of the slides, followed by data analysis, with specific reference to lymphoma. Using this method, scores for both the mean intensity of a marker of interest and the percentage positivity are generated to facilitate further quantitative analysis. Multiplexing minimizes sample utilization and provides spatial information for each marker of interest.
Source Title: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
URI: https://scholarbank.nus.edu.sg/handle/10635/207278
ISSN: 1940087X
DOI: 10.3791/58711
Appears in Collections:Staff Publications
Elements

Show full item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
Hong G. Jove 2019. MIF analysis in lymphoma.pdfPublished version3.44 MBAdobe PDF

CLOSED

Published

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.