ELISPOT assays provide a powerful, highly sensitive tool for gauging the production of cytokines and other cellular secretions; however, they have traditionally relied on visual spot counting. Modern ELISPOT reader technology allows for more precise and repeatable spot counting will less dependency on human judgment.
Some recent innovations include answering questions such as:
- Which cytokine spots are produced by antigen-specific T
cells (that are to be measured) vs. those produced by bystander cells
(being of no T cell diagnostic value)?
- How can the software designs be made both intuitive, yet flexible?
- What are the implications of small spots vs larger spots?
- How should the thresholds for minimum and maximum spot
size be set, when counting and analyzing the different cytokine ELISPOT
- In what frequency range are accurate measurements made?
- Can measurements be made at single-cell resolution?
- To what extent can antagonistic cytokines interfere with the results?
- How many replicates are required to produce significant
results? That is, how robust can these results be made using replicate wells?