Indian Muntjac – Deer Skin Fibroblast cells. Staining: Blue: DAPI, Green: Phalloidin-Alexa488, Red: Mitotracker CMXRos Top: image from full field of view; bottom: zoom in. Images by Jeroen Kole (Confocal.nl).
Comparison of performance of RCM and a PMT based confocal system
The resolution and sensitivity of the raw RCM image is better, resulting in a better deconvolution result.
The light intensity at the sample plane was measured at: 4.5 microwatt (1.0 AU), 12.5 microwatt (0.3 AU) and 3.0 microwatt (RCM).
Nuclear spread from fixed mouse spermatocytes, immunostained for SYCP3 a component of the synaptonemal complex (Alexa 488-labelling). Upper panel: images of a PMT based confocal with 1.0 AU pinhole, 0.3 AU pinhole and RCM image. Lower panel: same images deconvolved using Huygens Essential RCM module (SVI), using an experimental point spread function. Scalebar: 1 micrometer. Sample courtesy of A. Agostinho – Advanced Light Microscopy Facility, Science for Life Laboratory. Imaging and deconvolution performed by Jeroen Kole (Confocal.nl).
