ELISpot (Enzyme-Linked Immunosorbent Spot) is an immunoassay that measures the frequency of antigen-specific cytokine-secreting cells at the single-cell level. It builds on familiar immunoassay principles and is characterized by its high sensitivity. The assay requires a small number of cells,allows for multiple stimulations,and avoids the need for fixation or permeabilization,which simplifies the workflow and preserves cell function. Notably,ELISpot is a singleplex assay,meaning one cytokine can be analyzed per run.
"},{"faq_question":"How does FluoroSpot differ from ELISpot?","faq_answer":"FluoroSpot detects analyte-secreting cells using antibodies labeled with fluorophores rather than enzymes,which results in a one-step direct detection method compared to ELISpot’s two-step system. The key practical advantage of FluoroSpot is its ability to analyze multiple analytes in a single well,identifying cells producing each analyte individually as well as those producing both analytes simultaneously. This makes FluoroSpot well-suited to programs that require simultaneous cytokine profiling.
"},{"faq_question":"Can CellCarta customize ELISpot and FluoroSpot assays?","faq_answer":"Yes. CellCarta's team customizes both FluoroSpot and ELISpot tests according to the antigen stimulation selected and the cell type being investigated. The customization process follows a well-established validation framework,resulting in assays optimized to each sponsor's specific study design. Scientists have extensive experience developing assays for exploratory,secondary,and primary endpoint use in clinical trials.
"},{"faq_question":"What clinical applications do these assays support?","faq_answer":"FluoroSpot and ELISpot tests support a broad range of clinical applications,including monitoring the immunogenicity of vaccines,cell therapies,and immune-modulating therapies such as CAR-T cells,checkpoint inhibitors,cytokines,agonists,and adjuvants. They are also used for large-scale epitope mapping,detection of HLA-restricted responses,and measurement of memory B-cell and T-cell responses from PBMC samples. Across therapeutic areas,these assays are applied in cancer,infectious diseases,autoimmune conditions,and rare diseases.
"},{"faq_question":"When should I use ELISpot instead of ICS?","faq_answer":"Both ELISpot and intracellular cytokine staining (ICS) measure antigen-specific T cell responses,but each has distinct practical advantages. ELISpot typically shows higher sensitivity than ICS,making it well-suited for detecting low-frequency antigen-specific responses. It also requires fewer cells and avoids fixation and permeabilization steps. ICS,by contrast,provides information about the cellular subpopulation expressing the cytokines of interest and allows characterization of cytokine-producing T cell subsets by surface phenotype. The right choice depends on the study’s endpoint requirements and the information needed.
\nFor more insight,read our article on ICS vs ELISpot for clinical trials,and consult with our experts.
"}]}#-supreme-cellcarta-service-blocks-7-701{blocks-data:[{"image":"https://cellcarta.com/wp-content/uploads/2026/07/CellCarta_-37-2-2-scaled.jpg","image_alt":"Ella multiplex ELISA automated benchtop system","title":"Ella™:Multiplex ELISA","description":"Generate up to 256 results in under 90 minutes with automated benchtop ELISA multiplexing.","button_text":"Discover","button_url":"/capabilities/immunoassay-services/ella-multiplexing-elisa/","button_target":"_self"},{"image":"https://cellcarta.com/wp-content/uploads/2026/07/CellCarta_-28-2-scaled.jpg","image_alt":"Meso Scale Discovery MSD electrochemiluminescence platform","title":"Meso Scale Discovery® (MSD)","description":"Measure up to ten analytes in each plate well using serum or plasma samples as small as 25 μL.","button_text":"Discover","button_url":"/capabilities/immunoassay-services/meso-scale-discovery/","button_target":"_self"},{"image":"https://cellcarta.com/wp-content/uploads/2026/08/CellCarta_-50-1-scaled.jpg","image_alt":"Immune monitoring multiplex assay workflow","title":"Immune Monitoring","description":"Multiplex assays for immune response monitoring across the clinical trial continuum.","button_text":"Discover","button_url":"/capabilities/immune-monitoring/","button_target":"_self"}]}#-supreme-cellcarta-tab-content-4-701{oxy-supreme-cellcarta-tab-content-content-heading-tag:h3;oxy-supreme-cellcarta-tab-content-content-heading-size:default;oxy-supreme-cellcarta-tab-content-content-body-size:default;tabs-data:[{"tab_title":"Single-Cell Insight","content_title":"","content_body":"ELISpot and FluoroSpot provides a highly sensitive count of functional antigen-specific responses at the single-cell level.
\nThese assays require only a small number of cells,allows for multiple stimulations,and involves minimal cell manipulations —no fixation or permeabilization required.
","tab_image":"https://cellcarta.com/wp-content/uploads/2026/06/Single-Cell-Insight-50.jpg","image_alt":"Scientist analysing ELISpot plate at single-cell level"},{"tab_title":"Cytokine Detection","content_title":"","content_body":"ELISpot uses enzymatic action to measure the secretion of a single cytokine,uncovering critical information about cell-mediated immunity. All resulting SFU counts are standardized using the ImmunoSpot® S6 Universal analyzer.
\nFor example,CellCarta can easily monitor memory T cell responses using a fit-for-purpose ELISpot test for IFNγdetection. This assay is readily available for exploratory use and can be validated for secondary and primary endpoints.
","tab_image":"https://cellcarta.com/wp-content/uploads/2026/07/ELISpot-for-IFNγ-tab-13.jpg","image_alt":"ELISpot assay plate for IFN-gamma detection"},{"tab_title":"Multiplex Analysis","content_title":"","content_body":"FluoroSpot uses fluorophore-labeled antibodies for one-step direct detection,enabling simultaneous analysis of multiple analytes in a single well.
\nNot only does this technology allows users to identify cells producing each analyte individually,but it can also detect cells co-producing analytes simultaneously.
\nWe have validated several multiplex FluoroSpot assays for use,including:
\nCellCarta's scientists can customize both ELISpot and FluoroSpot assays according to the antigen stimulation selected and cell type investigated.
\nOur customization workflow follows a well-established validation process designed to produce optimized,reproducible assays. Whether your study calls for singleplex or multiplex design,our team adapts to your protocol rather than fitting your program to some standard template.
","tab_image":"https://cellcarta.com/wp-content/uploads/2026/06/Assay-customization-470.jpg","image_alt":"Scientist customizing ELISpot assay in laboratory"}]}