Our FluoroSpot products

FluoroSpot is used to detect and quantify protein-secreting cells. This immunoassay combines the sensitivity of ELISpot with the capacity to study the secretion of several analytes simultaneously.

In FluoroSpot, a cell suspension is added to the wells of a plate coated with one or more protein-specific capture antibodies. Proteins secreted by the cells are immediately captured by these antibodies during the entire stimulation period. The cells are removed and then tagged or biotinylated protein-specific detection antibodies are added followed by fluorophore-labeled secondary reagents.

The plate is then read using a FluoroSpot reader where each fluorescent spot corresponds to a responding analyte-secreting cell.
 

Due to its high sensitivity, FluoroSpot is ideal for identifying polyfunctional cells in e.g., vaccine or cancer research. The detection of different secretory profiles separates the responding cells into different functional groups. Looking at several proteins in parallel increases the possibility of identifying distinct cell populations in the same well. 

IFN-γ is often combined with IL-2, TNF-α, or Granzyme B in analyses investigating cytotoxic T cell responses against selected antigens. Stimulation is usually achieved with peptide pools covering the vaccine antigen. Check out all our FluoroSpot combinations here.

FluoroSpot excels in B cell profiling by capturing multiple immunoglobulin classes or isotypes directly upon secretion, enhancing the detection of specific antibody responses. The assay not only makes it easy to identify rare antigen-specific antibody-secreting cells (ASCs) with high sensitivity but also allows for the assessment of circulating memory B cells. Regularly employed in immunological research, B cell FluoroSpot is particularly valuable for analyzing the breadth and specificity of B cell responses following infection or vaccination. Learn more about B cell profiling here.

Like ELISpot, FluoroSpot is useful in assessing vaccine-induced immunity by quantifying antigen-specific T and B cell responses but giving a broader picture of the response by enumerating highly functional triple-secreting IFN-γ/IL-2/TNF-α cells or even multiple immunoglobulins in the same well. The assays allow researchers to gauge a vaccine's immunogenicity and potential protective effects, providing critical insights into their performance and function. Our white paper takes a deeper look into ELISpot's role in vaccine development, offering a detailed perspective on its application in shaping vaccine-mediated immunity.

Mabtech's FluoroSpot kits enable precise immune monitoring, capturing the activity of killer T cells through key markers like IFN-γ and Granzyme B simultaneously. This technology has been instrumental in advancing cancer immunotherapy research, offering insights into how the immune system targets and destroys cancer cells. Explore our white paper to see how FluoroSpot has facilitated this groundbreaking work, spotlighting the assay's critical role in the development of novel cancer treatments.