Use of Immunohistochemistry in Disease Detection

*Immunohistochemistry - The Ageless Biotechnology*

Procedures and mechanisms. Journal of Histochemistry and Cytochemistry.

[45] Yamashita S. Immunoelectron microscopy with the post-embedding method based on heat-induced antigen retrieval: Techniques and mechanisms.

Kenbikyo. 2014;**49**:124-131 (in

2014;**62**:584-597

Japanese)

**52**

**55**

**Chapter 3**

**Abstract**

*Edwin Roger Parra*

cancer prevention and treatments.

**1. Introduction**

Immune Cell Profiling in

Immunofluorescence and

Digital Analysis Approaches

**Keywords:** tyramide signal amplification, conventional IHC protocol, immunoprofiling, cancer tissues, image analysis, spatial analysis

During the last years, multiplex immunofluorescence (mIF) has emerged as a very powerful tool in multiple epitope detection to study tumor tissues. This revolutionary technology is providing an important visual technique for tumor examination in formalin-fixed paraffin-embedded specimens for a better understanding of tumor microenvironment, new treatment discoveries, cancer prevention, as well as translational studies. The aim of this chapter is to highlight the use of tyramide signal amplification methodology in mIF and image analysis to identify several proteins at the same time in one single tissue and their spatial distribution in different tumor specimens including whole sections, core needle biopsies, and tissue microarrays. This type of methodology associated with image analysis can perform high-quality throughput assay in translational research studies to be applied in

In the last years, novel and effective immunotherapies for patients with different tumor types are becoming clinically important, because of the remarkable clinical efficacy observed with several immune checkpoint inhibitors such as cytotoxic T lymphocyte antigen 4 (CTLA-4) and the programed death receptor 1 (PD-1) or its ligand (PD-L1) [1–12]. Whereas anti-CTLA-4 antibodies (ipilimumab and tremelimumab), anti-PD-1 antibodies (nivolumab and pembrolizumab), and anti-PD-L1 antibodies (atezolizumab, avelumab, and durvalumab) have produced remarkable results, increasing the survival prognosis in many cancer types, it is still unknown why some tumors do not respond to or relapse after this type of treatment. In this way, increased observations suggest that tumors rich in tumor-associated immune cells (TAICs) may respond to therapies targeting immune system inhibitory or stimulatory mechanisms, and tumors with non-TAICs may require additional interventions aimed at promoting optimal inflammation and innate immune activation in the tumor microenvironment [13–15]. Indeed, characterization of different immune checkpoints as well as tumor microenvironment in patients with cancer has become

Cancer Using Multiplex
