Anti-Fading Agents for Confocal Immunofluorescence: Colocalization of Nuclear Polypeptides

Abstract

We evaluated the performance of four anti-fading agents during acquisition of multiple optical sections near the widest diameter of Drosophila accessory gland nuclei using indirect immunofluorescence and confocal laser scanning microscopy. Two commercially available agents, Vectashield® and SlowFade® showed anti-fading properties that alleviated fluorochrome fading associated with the acquisition of multiple fluorescent optical Z-series from a single specimen by a confocal laser scanning system. Using these reagents, we were able to colocalize polypeptides through immunostained whole Drosophila nuclei.

Key Words:

• Drosophila
• nucleus
• histone
• immunocytochemistry
• fluorochrome
• confocal microscopy