Technical Reference #1896

1896.

Cellular MicroRNA and P Bodies Modulate Host-HIV-1 Interactions Robin Nathans; Chia-ying Chu; Anna Kristina Serquina; Chih-Chung Lu; Hong Cao; and Tariq M Rana, University of Massachusetts Medical School, Molecular Cell, 34(1896), (2009)
Link To Paper

Materials & Methods:
293T cells were cultured in 35 mm dishes with glass coverslip bottoms (Mat- Tek Corporation Ashland MA). Cells were transfected with expression vectors for YFP-tagged proteins using Lipofectamine 2000 according to the manufacturer’s protocol. Twenty-four hours later cells were fixed and monitored for YFP signals of transiently expressed proteins. Signals were detected by a Leica confocal imaging spectrophotometer system (TCS-SP2) attached to a Leica DMIRE inverted fluorescence microscope equipped with an argon laser two HeNe lasers an acousto-optic tunable filter (AOTF) to attenuate individual visible laser lines and a tunable acousto-optical beam splitter (AOBS). A 633 1.32 NA oil-immersion objective was employed.

Microscopic Technique
confocal imaging spectrophotometer system, inverted fluorescence microscope

Cell Type(s)
293T cells