Fluorospot flex

Manufactured by Mabtech

Sourced in Sweden

The FluoroSpot flex is a versatile lab equipment used for the detection and quantification of cytokine-secreting cells. It utilizes fluorescent-labeled antibodies to enable high-sensitivity and high-throughput analysis of cellular immune responses.

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Lab products found in correlation

Iris fluorospot reader, by Mabtech (1 mentions)

2 protocols using fluorospot flex

Quantifying Antibody-Secreting Cells

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The number of IgM, IgA, and IgG ASCs was determined using FluoroSpot flex (MABTECH AB, Sweden) following the manufacturer’s instructions. In brief, 5×10⁴ cells were added per well in 200 µl cell culture medium containing 10% fetal bovine serum. The cells were then incubated overnight in a 37°C humidified incubator with 5% CO2. Cells were then removed and detection antibodies were added to per well and incubated for 2 hours at 20°C. The spots were counted using a FluoroSpot reader (MABTECH AB, Sweden).

Huang T., Pi C., Xu X., Feng Y., Zhang J., Gu H, & Fang J. (2024). Effect of BAFF blockade on the B cell receptor repertoire and transcriptome in a mouse model of systemic lupus erythematosus. Frontiers in Immunology, 14, 1307392.

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Quantifying SARS-CoV-2 T Cell Responses

Cited 1 time

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To quantify SARS-CoV-2 specific CD4 and CD8 T cells, peptide restimulation with Fluorospot for IFN and IL-2 was performed (Mabtech; Fluorospot Flex). In short, 2,5X10ˆ5 PBMC were resuspended in complete medium, plated in triplicate into 96 well plates with a PVDF membrane bottom layer, precoated overnight with capturing antibodies directed against IFN and IL-2. PBMC were stimulated with CD4 T cell (CD4-R and CD4-S MP) or CD8 T cell (CD8-A and CD8-B MP) specific peptide pools at a final concentration of 1 mg/mL, as described in Weiskopf et al.⁵⁴ (link) After 23 hours of stimulation, plates were collected and spots were developed following manufacturer’s protocol (Mabtech, FSP-0102-10). Spots were revealed and quantified using Mabtech IRIS Fluorospot reader (Mabtech).

Bosteels C., Van Damme K.F., De Leeuw E., Declercq J., Maes B., Bosteels V., Hoste L., Naesens L., Debeuf N., Deckers J., Cole B., Pardons M., Weiskopf D., Sette A., Weygaerde Y.V., Malfait T., Vandecasteele S.J., Demedts I.K., Slabbynck H., Allard S., Depuydt P., Van Braeckel E., De Clercq J., Martens L., Dupont S., Seurinck R., Vandamme N., Haerynck F., Roychowdhury D.F., Vandekerckhove L., Guilliams M., Tavernier S.J, & Lambrecht B.N. (2022). Loss of GM-CSF-dependent instruction of alveolar macrophages in COVID-19 provides a rationale for inhaled GM-CSF treatment. Cell Reports Medicine, 3(12), 100833.

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