Table 6. Autofluorescence and specific fluorescence at peak of emission for Cy3 & Cy5 excited by white light LED (SOLA), PE2 (535nm & 615nm) or PE4000 (555 & 635nm).

Cy3 at 570nm	PE2 535nm		PE4000 555nm		Sola
	BK	specific	BK	specific	BK	specific
average	1.989757	9.160652	1.588668	6.795557	1.327651	5.540776
sem	0.253373	0.182925	0.014931	0.123194	0.224278	0.105282
ratio specific/bk		4.603906		4.277519		4.173367
Cy5 at 670nm	PE2 615nm		PE4000 635nm		Sola
	BK	specific	BK	specific	BK	specific
average	0.272805	0.919322	0.26374	1.122252	0.128934	0.52169
sem	0.01203	0.022824	0.025742	0.02747	0.010783	0.014581
ratio specific/BK		3.369884		4.255147		4.046169

The specific staining was quantified in thresholded positive cells and 3 regions of interest were drawn manually in unstained tissue for quantification of the background fluorescence (BK).

The average fluorescence intensity (counts/s) was quantified at the peak of fluorescence on images taken at the appropriate wave length by the Nuance camera with Tunable filter (ie 570nm for Cy3 & 670nm for Cy5).

Peaks of excitation for Cy3 & Cy5 are 554nm & 649nm respectively.