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. Author manuscript; available in PMC: 2014 Jan 1.
Published in final edited form as: Methods Mol Biol. 2013;931:29–59. doi: 10.1007/978-1-62703-056-4_2

Fig. 3.

Fig. 3

Images of BPAE cell slide from Molecular Expressions labeled with DAPI, AlexaFluor® 488 Phalloidin, and MitoTracker® CMXRos. Images were collected on an Olympus IX71 microscope coupled with a mercury HBO lamp and using DAPI, FITC, and Texas Red BrightLine® hard coated filter sets (Semrock). Images were collected on a Retiga 2000R camera at full resolution with the same settings using three different 20× lenses. The lamp intensity was attenuated to 6%, with exposure times of 250 ms for DAPI, 1,000 ms for AlexaFluor® 488, and 500 ms for MitoTracker® used. Lenses used were (a) Plan 20×/0.4 NA; (b) UPlanFL N 20×/0.5NA; (c) UPlanSApo 20×/0.75NA. Images of the same sample as in (a–c) were collected using a 40×/0.6NA LWD, U PLAN FLlens with a coverslip thickness correction collar. Images were collected with the collar set correctly to 0.17 mm (d) or incorrectly to 1 mm (e). The image contrast, brightness, and gamma factor were all adjusted to the same levels for the images displayed in (a–c) as well as and for (d, e). The gamma factor was used to bring up dim features so they are visible within the images by eye. Scale bar is 10 μm.