Protocols in apoptosis identification and affirmation

Abstract

Protocols are set of predefined written procedural documents that offer guidance or specifications for performing the experiments and its implementation. They are an essential component of good laboratory practice (GLP) and good clinical practice (GCP) regulations that establish the study’s rationale, objectives, design, and planned analyses of the results, as well as the conditions under which these procedures will be carried out. In addition to detailed procedures, protocols often include information on safety precautions, the calculation of results, and reporting standards, including statistical analysis and rules for predefining and documenting excluded data to avoid bias.

The protocol is the heart of every experiment. It is the plan; it is a critical document for everyone involved in the conduct of the procedure—Rebecca Kush, President of CDISC

Different protocols are available which provide signaling pathways of introduction to “Programmed cell death” which called “Apoptosis”. The course of programmed cell death or “Apoptosis” is generally regarded as a typical element of the human health and disease state. Normal homeostasis is sustained throughout by maintaining balance between cell death and cell division. Apoptosis is phenomenon that involves specific activation of death-signaling pathways leading to the removal of cells from tissue. The individual morphological features of apoptosis include cell shrinkage, chromatin condensation, membrane blebbing, and formation of apoptotic bodies. Inapt apoptosis is a key factor in many diseased conditions, including neurodegenerative diseases, cancer, I-R injury, and many more. In addition, the area of apoptosis research is growing at an alarming rate and till date vast research has been made on the clarification and analysis of the cell cycle mechanisms and signaling pathways regulating cell cycle. The aptitude to detect and quantify apoptosis and to understand its biochemistry related to regulatory genes and proteins is highly crucial for biomedical research. This chapter is an attempt to highlight various basic as well as advanced tools and techniques (like immunohistochemistry, TUNEL assay, in-situ end-labeling techniques in conjunction with standard flow cytometry) used in area of apoptosis research in addition to focus on protocols used in qualitative and quantitative determination of apoptosis to be followed during preclinical and clinical phase of drug development. The use of aforementioned techniques for apoptosis quantification will enable clinical investigators to accurately assess apoptosis in context of various diseases.