The perk of RAWspot™ – our algorithm

Originally published October 13, 2022, updated December 20, 2022


Besides intuitive software and plug-and-play hardware, the most striking feature of our readers is their novel spot counting algorithm, RAWspot.

 

 

Exact spot centers

The power of RAWspot™ lies in its unparalleled ability to accurately detect spot centers in all wells analyzed. This is an essential factor in FluoroSpot, as location of the center point forms the basis identifying cells secreting multiple analytes.

RAWspot center zoom

Exact spot centers ensure reliable multiplexing.

RAWspot in ELISpot

For FluoroSpot as well as ELISpot

Despite no need for multiplexing, the algorithm excels also in ELISpot. Thanks to the accuracy in spot detection, our readers easily identify distinct spots in a linear fashion even with more than 3000 spots per well.  Thus, regardless if you are using enzymatic or fluorescent based detection, RAWspot consistently detects every spot, also in positive control wells.

 

 

So how does it work?

The algorithm processes the wide dynamic range of the image RAW signal in contrast to analysis of flat 8-bit images, like JPEG or TIF. Image RAW represents the uncompressed signal straight from the camera sensor, thus avoiding the problem of over-saturated spots.

Using the increased processing power of modern graphics cards, RAWspot™ extracts a 3-dimensional volume for each spot, corresponding to the relative amount of secreted analyte. This new dimension of spot-data is called Relative Spot Volume (RSV).

Read more in this technote on how RSV values can be used for a more detailed characterization of T cell responses.

For nitty gritty details, check out the this video, where the creator of the algorithm, Professor Joakim Jaldén at the KTH Royal Institute of Technology, breaks down the science behind RAWspot.

Finally, if you are interested mathematical research behind RAWspot™, two scientific publications by Professor Joakim Jaldén and his student Dr. Pol del Aguila Pla, can be found here and here.


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