OPAL Multiplex Immunofluorescence (mIF) Protocol: 4-Plex Staining for CD155, GPNMB, CD31, and CD112 v1

2026 Preprint Open access

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Authors: Franz Zemp, Hongrui Lui, Douglas Mahoney
DOI: 10.17504/protocols.io.eq2lymznmlx9/v1
Cited by: 1

Abstract

Simultaneous detection and quantification of multiple protein targets in formalin-fixed, paraffin-embedded (FFPE) tissue sections using the Opal 6-Plex Manual Detection Kit (Akoya Biosciences). This protocol enables multiplex immunofluorescence staining for CD155, GPNMB, CD31, and CD112 with tyramide signal amplification (TSA) and spectrally distinct fluorophores.

Fields of Study & Subjects

Single-cell and spatial transcriptomics Advanced Fluorescence Microscopy Techniques Peptidase Inhibition and Analysis

Publication details

Publication date
27 May 2026
Document type
Preprint