ELISPOT assays allow visualization of the secretory product of individually activated or responding cells.

The cells are cultured and stimulated on plates coated with antibodies that capture specific target molecules released by the activated cells. The presence and distribution of these analytes is then detected through a sandwich ELISA that deposits a reporter substance where the analyte is capture.

Each spot that develops in the assay represents a single reactive cell, providing both qualitative (type of immune protein) and quantitative (number of responding cells) data.


FLUOROSPOT assays employ sandwich ELISA technology, with fluorescence detection instead of chromogenic detection.

Used in the analysis of cytokines and other soluble molecules produced from secreting T-cells, FLUOROSPOT can identify secreted products at a single cell level.

Learn more about dual staining

Dual ELISPOT and FLUOROSPOT can detect secreted cytokines and measure the frequency of single cells that simultaneously produce two cytokines or other effector molecules. They help you save cell material and measure cytokines that are co-expressed.

Dual ELISPOT and dual FLUOROSPOT differ by theirt detection methods

  • Dual ELISPOT
    • contain ready-to-use AEC substrate buffer (red spots) and BCIP/NBT substrate buffer (blue spot)
    • contain anti-FITC antibody green fluorescene and streptavidin-phycoerythrin conjugates

Click and expand the image below to learn more.

Colorimetric detection diagram