Longitudinal assessment of fluorescence stability shows fluorescence intensity decreases over time: implications for fluorescence microscopy studies
Fig 4
Decreased fluorescence intensity of OE sections triple-stained for OMP-AF488, GAP43-AF546, and EdU-sulfo-Cyanine5 over the course of weekly confocal microscopy.
Sections were triple-stained with anti-OMP primary/AF488-conjugated secondary, anti-GAP43 primary/AF546-conjugated secondary and EdU (click chemistry detection with sulfo-Cyanine5). (a-c) MIPs of anti-OMP primary antibody with AF488-conjugated secondary antibody-stained OE section imaged with confocal microscopy. (d) Significant decrease in AF488 fluorescence intensity in single optical sections. (e) Significant decrease in AF488 fluorescence intensity in MIPs. (f-h) MIPs of anti-GAP43 primary antibody with AF546-conjugated secondary antibody-stained OE section imaged with confocal microscopy. (i) Significant decrease in AF546 fluorescence intensity in single optical sections. (j) Significant decrease in AF546 fluorescence intensity in MIPs. (k-m) MIPs of EdU with sulfo-Cyanine5 fluorophore-stained OE section imaged with confocal microscopy. (n) Significant decrease in sulfo-Cyanine5 fluorescence intensity in single optical sections. (o) Significant decrease in sulfo-Cyanine5 fluorescence intensity in MIPs. Images of weeks 1, 3 and 5 are shown as examples. Each line of data represents an individual mouse.