US EPA

1801340 

Journal Article 

COMPARISON OF ANTI-FADING AGENTS USED IN FLUORESCENCE MICROSCOPY - IMAGE-ANALYSIS AND LASER CONFOCAL MICROSCOPY STUDY 

Longin, A; Souchier, C; Ffrench, M; Bryon, PA 

1993 

Yes 

Journal of Histochemistry and Cytochemistry
ISSN: 0022-1554
EISSN: 1551-5044 

41 

12 

1833-1840 

English 

8245431 

To ascertain the ability of commercial and home-made anti-fading media to reduce the decrease of fluorescein isothiocyanate (FITC) fluorescence, we studied the bleaching characteristics of FITC-stained Reh 6 cells mounted in buffered glycerol and in anti-fading media. We measured the intensity of fluorescence over time with a confocal laser scanning microscope and a standard epifluorescence microscope coupled to an image analysis system. Most of the anti-fading media effectively retard fading but each has drawbacks. Better results were obtained with media containing p-phenylenediamine (solutions in buffered glycerol, Vectashield, Fluorstop). However, Mowiol, Slowfade, n-propyl gallate (20 g/liter) were also effective in retarding fading. Most of them, except Mowiol, reduced fluorescence intensity. We concluded that the choice of anti-fading medium would depend on the desired results: a slower decay of fluorescence despite an initial quenching of fluorescence or a lower retardant effect with no decrease in initial fluorescence intensity. Moreover, the combination of Mowiol with another anti-fading medium may be a useful compromise when a strong retardant effect is required without marked quenching of the initial fluorescence. 

FADING; FLUORESCENCE; ANTI-FADING; FLUORESCEIN ISOTHIOCYANATE (FITC); RHODAMINE B ISOTHIOCYANATE; IMAGE ANALYSIS; CONFOCAL MICROSCOPY