1. A method of fluorescent dot counting in an image programmed in a memory of a device comprising: a. determining dot candidate seeds, wherein determining the dot candidate seeds comprises: i. applying a tophat transform to the image; ii. applying h-maxima suppression; iii. detecting regional maxima; and iv. performing connected component analysis; b. segmenting dot candidate patches; c. extracting dot candidate features; and d. classifying dot candidates.