Retrofitted confocal laser scanner for a commercial inverted fluorescence microscope

We describe the design and implementation of an inverted laser scanning confocal fluorescence microscope utilizing the commercial Nikon Diaphot TMD platform. An external confocal scanner was retrofitted through the video side port of the Diaphot. With 10x, 0.5 NA dry and 60x, 1.4 NA oil immersion objectives, the depth discrimination is 5.8 mum and 0.8 mum, respectively, as determined by derivatives of fluorescence edge responses measured in liquid samples of rhodamine 6G dissolved in DMSO. We present sample edge response curves and representative confocal fluorescence images of tumor cells in monolayer culture. (C) 2001 American Institute of Physics.