ASC Fluorescence and Brightfield Imaging

First step of the system check. The purpose is to test the general performance of the system: Do the basic functions work fine? Does the acquisition software behave as it should? Can one set up Koehler illumination? Are the transmitted and the reflected beam-paths ok? Although this test does not represent high-resolution work, it might reveal general problems with the system.

requirements

• basic sample with fluorescence and contrast in transmitted light (i.e. Zeiss/Leica Convallaria demo sample)

acquisition

• set up the beam-path for fluorescence and transmitted light
• use basic air objective (i.e. 10x/0.4)
• acquire two images in basic resolution (i.e. 512x512)

analysis

• Does the illumination look reasonably homogeneous over the whole field of view in both images?
• Is the transmitted light image free of dirt particles?
• Are both images free of artifacts?

images

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  Convallaria, Zeiss LSM 510, 10x/0.45 objective, 488 nm excitation, BP505-550 detection, pinhole: 1 airy unit

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  Convallaria, Zeiss LSM 510, 10x/0.45 objective, 488 nm illumination, transmitted light detection