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Aid ispot reader

Manufactured by Autoimmun Diagnostika
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Sourced in Germany

The AID iSpot reader is a compact and automated instrument designed for the detection and analysis of cytokine-secreting cells. It utilizes an ELISPOT-based technology to quantify the frequency of antigen-specific T-cells or other immune cells in biological samples.

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Lab products found in correlation

Ifn γ antibody, by R&D Systems (2 mentions) Streptavidin alkaline phosphatase, by Merck Group (2 mentions) Complete rpmi medium, by Merck Group (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Cpg odn 1826, by InvivoGen (1 mentions) Pvdf 96 well plates, by Merck Group (1 mentions) Pvdf plate, by Merck Group (1 mentions) Anti cd8α, by BD (1 mentions)

Spelling variants of this product

AID iSpot (7 citations) AID iSpot Reader System (7 citations) ISpot Reader (2 citations) AID iSpot ELISpot reader (2 citations)

4 protocols using aid ispot reader

1

Quantifying Antigen-Specific Plasma Cells

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For analysis of memory B-cells, splenocytes and lung cells were stimulated (5 × 10E5 cells per well, 24-well plate; 5 days, 37 °C) with 10 μg/mL CpG ODN 1826 (Invivogen, San Diego, CA), 10 μg/mL pokeweed mitogen (Sigma-Alderich, Zwijndrecht, The Netherlands), and Staphylococcus aureus protein A of Cowan Strain (1:5000; Sigma) in RPMI complete medium with β-mercaptoethanol (1:25000; Sigma) to induce differentiation into antibody secreting cells24 (link). The percentage of OMV specific antibody secreting cells were subsequently determined by ELISpot. The numbers of OMV-specific IgG- and IgA-producing plasma cells in blood, spleen and lungs were directly determined using the same ELISpot method, with 10 µg/mL wildtype B1917 OMV as coat, as described before15 (link). Spots were counted with an AID iSpot reader (Autoimmun Diagnostika, Strassberg, Germany) and indicated as antibody-secreting cells per 5 × 10E5 cells.
Raeven R.H., Rockx-Brouwer D., Kanojia G., van der Maas L., Bindels T.H., ten Have R., van Riet E., Metz B, & Kersten G.F. (2020). Intranasal immunization with outer membrane vesicle pertussis vaccine confers broad protection through mucosal IgA and Th17 responses. Scientific Reports, 10, 7396.
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2

Detection of OMV, BrkA, and Vag8-specific Plasma Cells

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For the detection of OMV, BrkA or Vag8-specific IgG-producing plasma cells in spleen, filter plates were coated with 10 µg/mL WT B1917 OMV, 5 µg/mL BrkA or 5 µg/mL Vag8 and the ELISpot method was used as described before [20 (link)]. Spots were counted with an AID iSpot reader (Autoimmun Diagnostika, Strassberg, Germany) and indicated as antibody-secreting cells (ASC) per 5 × 105 cells. For analysis of memory B-cells, splenocytes were stimulated to induce differentiation into antibody-secreting cells as described previously [21 (link)]. The amounts of OMV, BrkA and Vag8-specific ASC were subsequently determined by using the same ELISpot method, only here 1 × 105 stimulated cells were added to the coated plates.
Raeven R.H., van Vlies N., Salverda M.L., van der Maas L., Uittenbogaard J.P., Bindels T.H., Rigters J., Verhagen L.M., Kruijer S., van Riet E., Metz B, & van der Ark A.A. (2020). The Role of Virulence Proteins in Protection Conferred by Bordetella pertussis Outer Membrane Vesicle Vaccines. Vaccines, 8(3), 429.
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3

Analysis of Tumor-Specific CD8α+ T-cells

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For analysis of tumour antigen-specific CD8α+ T-cell response, ELISPOT assay was performed with splenocytes from immunized mice. Spleens were collected aseptically, processed into single-cell suspension, and plated with 5 × 105 splenocytes per well in 96-well PVDF plates (EMD Millipore) pre-coated with IFN-γ antibody (R&D Systems) overnight. Splenocytes were then restimulated with antigen peptides (2 μg ml−1) or controls for 24 h. Assays were completed using sequential incubations with biotinylated secondary antibody, streptavidin alkaline phosphatase (Sigma Chemical) and NBT/BCIP substrate (Surmodics). Developed spots were analysed using an AID iSpot Reader (Autoimmun Diagnostika GmbH).
Son S., Nam J., Kim A.S., Ahn J., Park K.S., Phoo M.T., Sherren B., Zou W., Lee S.H., Farokhzad O.C., Shi J, & Moon J.J. (2022). Induction of T-helper-17-cell-mediated anti-tumour immunity by pathogen-mimicking polymer nanoparticles. Nature biomedical engineering, 7(1), 72-84.
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4

Analyzing Tumor-Specific CD8+ T-cells

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For analysis of tumor-antigen-specific CD8α+ T-cells in the systemic circulation, submandibular bleeding was performed at indicated time points. Red blood cells in PBMCs were lysed with ACK lysis buffer twice at room temperature, followed by two washing steps with FACS buffer. PBMCs were blocked with CD16/32 antibody for 10 min and then stained with H-2Kb OVA tetramer-SINNFEKL (MBL International) and anti-CD8α (BD Biosciences) for flow cytometry analysis. ELISPOT assay was performed with splenocytes from immunized mice. Splenocytes were harvested aseptically, processed into a single-cell suspension, and plated with 5 × 105 splenocytes per well in 96-well PVDF plates (EMD Millipore) precoated with IFN-γ antibody (R&D Systems) overnight. Splenocytes were then restimulated with antigen peptides (2 µg/mL) or controls for 24 h. Assays were completed using sequential incubations with biotinylated-secondary antibody, streptavidin alkaline phosphatase (Sigma Chemical), and NBT/BCIP substrate (Surmodics). Developed spots were analyzed using an AID iSpot Reader (Autoimmun Diagnostika GmbH, Germany).
Son S., Nam J., Zenkov I., Ochyl L.J., Xu Y., Scheetz L., Shi J., Farokhzad O.C, & Moon J.J. (2020). Sugar-Nanocapsules Imprinted with Microbial Molecular Patterns for mRNA Vaccination. Nano letters, 20(3), 1499-1509.
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