Method and system for analysing fluorospot assays

A system for analysing fluorospot assays, the system comprising:

• (i) a light source arrangement configured to generate a plurality of excitation lights, wherein at a given time, the light source arrangement generates only one excitation light;
• (ii) a collimation arrangement positioned on an optical path of a generated excitation light, wherein the collimation arrangement is configured to collimate the generated excitation light;
• (iii) a multiband beam splitter positioned on an optical path of the collimated excitation light, the multiband beam splitter being supported by a support arrangement, wherein the multiband beam splitter is configured to:
• (a) reflect the collimated excitation light onto an assay plate for illuminating a well of the assay plate having a plurality of wells;
• (b) receive a reflection of the excitation light and an emitted light from the assay plate;
• and
• (c) reflect a portion of the reflected excitation light whilst transmitting a remaining portion of the reflected excitation light and the emitted light to a filtering arrangement;
• (iv) the filtering arrangement comprising a filter wheel and at least one emission filter, wherein the filter wheel is configured to accommodate the at least one emission filter therein, and wherein the at least one emission filter is configured to
• (d) remove the remaining portion of the reflected excitation light received from the multiband beam splitter;
• and
• (e) purify the emitted light whilst transmitting the emitted light to an optic arrangement;
• (v) the optical arrangement configured to direct the purified emitted light onto an imaging device;
• (vi) the imaging device configured to capture at least one image of the well, in raw image format, for each of the plurality of excitation light;
• and
• (vii) a processing module coupled to the imaging device, wherein the processing module is configured to
• (f) generate a model of analyte release distribution in the well for each of the plurality of excitation lights by optimizing a pre-diffusion analyte distribution based on the detected at least one potential release site;
• wherein optimizing the pre-diffusion analyte distribution is implemented iteratively at least once, and comprises employing at least one of:
• alternating direction methods, multiplicative update rules, forward-backward proximal gradient algorithms;
• and
• (g) for the plurality of excitation lights employed to illuminate the well of the assay plate, cluster a plurality of co-positioned fluorospots as a multiple secretion fluorospot, wherein the clustering is performed for all generated models of analyte release distribution, and wherein the clustering determines at least one multiple secretion fluorospot.