ELISpot Assay
The Enzyme-Linked ImmunoSpot (ELISpot) assay is a flexible and sensitive method for investigating the frequency of cytokine-secreting cells within a population. In virology and vaccine research, this assay provides a useful format for evaluating the T cell response to viral peptides. The Cytation™ 7’s upright color camera rapidly captures whole well images of ELISpot samples that are then analyzed using an automated Gen5™ protocol to quantitate immune response to antigenic stimuli. This method has also been optimized for two-color ELISpot formats relying on two capture antibodies with independent color developers to quantify the number of cells secreting two distinct cytokines.
ELISpot Assay Procedure
Capture
Samples are imaged using the upright color camera, with whole-well images stitched and processed to reduce background and improve contrast.
Analyze
All spots are identified using green signal. For two-color formats, the ratio of red and blue intensity are then used to identify spots corresponding to the secretion of the two separate cytokines (yellow and fuchsia masks).
Report
Total number of spots and subpopulation statistics for multiple cytokines are automatically reported.
Increase assay throughput and performance
Liquid handling automation
The BioTek 405™ TS or other Plate Washer can be used to automate the tedious wash steps required for processing ELISpot plates.
BioStack™ Compatible Assay:
Process up to 50 microplates
Incorporating the BioStack Microplate Stacker enables processing and analysis of up to 50 plates.
Application Notes
Visual Abstracts
Using ELISpot to Monitor Stimulation of PBMCs
Webinars