Related Experiment Video
Updated: Mar 30, 2026
Enzyme-linked Immunospot Assay ELISPOT: Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens
Published on: November 23, 2010
Enzyme-Linked ImmunoSpot (ELISpot) for Single-Cell Analysis
Sylvia Janetzki1, Rachel Rabin2
1ZellNet Consulting, Inc., 555 North Avenue, Suite 25-S, Fort Lee, NJ, 07024, USA. sylvia@zellnet.com.
Insights
The Enzyme-Linked ImmunoSpot (ELISpot) assay detects low-abundance analytes by capturing secreted molecules from single immune cells. This robust technique quantifies cellular responses and is expanding to analyze polyfunctional cells.
Area of Science:
- Immunology
- Biotechnology
- Cellular Biology
Background:
- The Enzyme-Linked ImmunoSpot (ELISpot) is a heterogeneous immunoassay crucial for detecting low-abundance analytes.
- It enables monitoring of immune system responses at the single-cell level by capturing secreted molecules.
Purpose of the Study:
- To describe optimized protocol steps for the ELISpot technique.
- To highlight improvements and tools for the scientific community.
- To discuss the expansion of ELISpot into polyfunctional cell analysis.
Main Methods:
- Utilizes membrane-bound antibodies to capture secreted molecules from individual cells.
- Employs ELISA-related development steps to visualize captured molecules.
- Generates distinct spots representing cytokine-secreting cells for quantification.
Main Results:
- Provides quantification of cells secreting specific molecules.
- Offers insights into the kinetics and strength of cellular secretion.
- Demonstrates the adaptability of ELISpot for advanced cellular analysis.
Conclusions:
- The ELISpot assay is a reliable and robust method for single-cell immune monitoring.
- Optimized protocols and available tools enhance its utility.
- The technique's expansion into polyfunctional analysis broadens its applications in immunology research.
Abstract:
The ELISpot, a heterogeneous immunoassay, is widely used for detection of low abundant analytes. It is a reliable and robust assay to monitor responses of the immune system at the single-cell level by capturing secreted molecules of interest with specific, membrane-bound antibodies. Those molecules are then made visible by a cascade of ELISA-related development steps. The final results are distinct spots on the membrane as an imprint of the cell secreting the captured molecules, not only allowing their quantification but also providing insight on the kinetics and strength of secretion. This chapter describes the optimized protocol steps of the ELISpot technique, important improvements and tools available for the community, and the current expansion of the technique into polyfunctional cell analysis.
More Related Videos
08:26The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity
Published on: December 14, 2016
13:41Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus
Published on: March 8, 2012
