•Enzymatic Elispot labeling
•allows to label 1-2 cytokines per cell synchronous.
•If you are interested to see ‘double secreting’ cells the stain of two cells in neighborhood may overlap. So, It is not clear if two cells are overlapping or one cells is really secreting two different cytokines ( see below left)

‘Fluorospot’ labeling
Allows to label a (theoretical unlimited number different cytokines) Because excitation for each cytokine is applied -one after the other-, decisions for double secreting cells is clearly.
One Image is acquired for each cytokine!

Currently Fluorospot assays with 1-4 Fluorophores are on the market.

Fluorospot offer's significant advantages over colorimetric formats, particularly in the areas of multiplexing and automated spot detection.

Moreover, as spot development is not enzymatic, signal intensity is directly proportional to the amount of analyte within.