FluoroXTM Fluorospot Analysis Software Suite
For the individual colour planes, FluoroXTM follows the same rules for the scientifically validated identification of spots (link to Wiki Page 21) as does the single-colour ImmunoSpotTM Software. Rigorous experimental validation has shown, however, that for the identification of multi-analyte positive spots (generated e.g. by cytokine co- expressing T cells) precise centers of spot overlap algorithms are NOT suited: instead, Center of Mass Distance Algorithms need to be used, and are used by FluoroX, to accurately identify cytokine co-expressing individual T cells compensating for cells movement during the analyte secretion period(cited reference). (we need to get copyright and create link to the full article). 21 CFR Part 11 compliant QC and audit trails support transparent fluorescent counting in stricter environment.
The FluoroXTM platform implements multiple image formats. For high-throughput multi-color counting, where quick turn-around time and generating large amounts of data are limiting factors, a color resolution of 8 bit RGB images (256 levels of intensity per channel) is generally more than adequate for accurate multi-channel FluoroSpot assay analysis. For high-content quantitative analysis of multi-color FluoroSpot data, raw color 16 bit images with a linear pixel intensity function (65,536 levels of intensity per channel) are available. When an extremely wide (more than 1000 X difference) dynamic range of fluorescence detection is required, the ImmunoSpot® Fluoro-X platform can be set to scan and count floating point 32 bit per channel High Dynamic Range (HDR) images with an infinite number of possible intensity levels.