CTL ImmunoSpot S5 core

ELISpot is one of the most sensitive mean to detect cytokine production from a single cell. A technology adapted from ELISA, the ELISpot utilizes membrane bound capturing antibody to capture cytokines of interest from cell that are culturing on the membrane. As soon as the cytokine are produced, it is captured locally and enriched around the cytokine-producing cell. At the end of the incubation, typically 24-48 hours, the cells were washed away and detecting antibodies and colorimetric substrate are subsequently applied. The end result is a visible spot formed around each individual cytokine producing cell. Thus the number of spot is a direct representation of number of cytokine-producing cells, and the area and intensity of each spot correlates with the amount produced. This assay practically allows the use of cytokine production as a report of T cell activation. Because of its sensitivity, ELISpot assay is particular suitable for detecting low frequency events or when working with sample with limited supply. Since a typical ELISpot assay is performed on plate following the standard 96-well plate format, any ELISpot assay can easily be adapted to high throughput format.  CTL ImmunoSpot S5 core

The use ELISpot is not restricted to cytokine detection. It has also been used for the detection of antigen-specific antibody as well as other secreted proteins and molecules. ELISpot has been used widely for frequency determination of antigen specific T/B cells, quantitative measurement of T cells response, Th1/Th2/Th17 immune response characterization, epiptope mapping, drug and vaccine development, auto-antigen/allergen screen and detection, clinicals test and bio-assay development, and more.

The CTL Immunospot S5 core is a robust instrument specifically designed for capturing and analyzing results from ELISpot assays. The high quality color CCD camera and light sources from above and below the plate ensure excellent image quality with capability to detect spot of 25 μm or smaller, with up to two colors. The S5 core is fast too, a typical 96-well plate is captured in less than 2 minutes. The instrument software features a user friendly and intuitive interface which makes data capturing a breeze. The analytical software is precise and versatile, allowing users to fine tune analytical parameters and perform data validation. The data export capability allows user to transfer professional looking results to common word processing or spreadsheet software for additional downstream processing.

A quick comparison between ELISpot, ELISA, suspension bead array (SBA, e.g. Luminex and CBA assay) and FACS analysis of intracellular cytokine staining for measuring cytokines production. For detail comparison of ELISpot to the other applications, please click here.

Question
ELISA SBA Flow cytometry ELISpot
Frequency of cytokine producing cells No  No Yes Yes
Lowest detection range N/A N/A 1 in 10e3-10e4  1 in 10e3-10e6
Measure secreted cytokine Yes Yes Yes/No Yes
Adaptability to high throughput format Yes Yes Yes/No Yes

Using ImmunoSpot S5 core analyzer

The ImmunoSpot S5 core analyzer is an user operatable instrument and requires some initial training on both image acquisition and analysis. We only provide the instrument for result acquisition. Please contact the core facility manager for demonstration, trial run, training or instrument use.

Fee

 Cost / plate
SCIID $10
TUFTS/TMC  $12.5
Non-Profit  $15
Commercial  $20

Image analysis using the CTL ImmunoSpot software is currently free of charge.

Key Specifications

  • High resolution color digital camera (2048 x 1536)
  • High quality lens optimized for distortion-free image capturing of entire well surface
  • High-precision two-axis automated stage.
  • Scan a 96-well plate in less than 2 minutes.
  • CTL ImmunoSpot V.5 software (Academic version
Please visit the manufacturer website for additional information.