Longitudinal assessment of fluorescence stability shows fluorescence intensity decreases over time: implications for fluorescence microscopy studies
Fig 1
Breakdown of analyses completed for each experiment using Control Exp. 1 (anti-OMP primary, AF488-conjugated secondary) as an example.
(a-c) Example images of OB section used for Control Exp. 1 (anti-OMP-AF488). (a) Image without overlays. (b) Same image with overlaid ROI used to assess combined background + staining fluorescence. (c) Same image with overlaid ROI used to assess background fluorescence (i.e., external plexiform layer (EPL)), in which no OMP staining is present. (d) Change in raw mean gray fluorescence values for total, i.e., background + stain (circle), stain (square), and background (triangle). (e) Percentage normalized mean gray fluorescence for total (combined background and stain) fluorescence. (f) Percentage normalized mean gray fluorescence for background fluorescence. (g) Percentage normalized mean gray fluorescence for staining fluorescence signal (total – background). Each line of data represents a single mouse.