Advanced system check
From BioDIP
MPI-CBG LMF
Advanced System Check
Contents |
Protocol (draft)
- fluorescence and bright field imaging (all systems)
- purpose: test general performance of the system - basic functions work fine? + Koehler illumination, light paths ok?
- requirements: Zeiss/Leica Convallaria demo sample
- work flow: acquire two images in basic resolution (512x512), blue ex / green em plus transmitted light image, using 10x objective
- field of illumination
- purpose: check for even illumination
- requirements: mirror
- work flow: 80/20 mirror, 488 nm laser, pinhole 1 AU. focus on reflective side of mirror. try to obtain "japanese flag".
- psf
- purpose:
- requirements:
- work flow:
- overlay UV/V and VIS (confocal systems with separate UV/V laser coupling)
- purpose: check the coupling precision of the UV/V laser
- requirements: 0.2/0.5µm fluorescent beads sample
- work flow: open pinhole(s), use high resolution apochromatic lens (NA 1.2 or above), acquire two images (UV/V plus VIS channel) with good sampling (pixel size ~100nm)
PSF Macro
install LUT
Mac: go to applications folder > fiji: ctrl+click on the Fiji(.app) icon > Show package content create "luts" folder if it's not there yet drag and drop the *.lut into it (re)start fiji
install macro
Fiji > Plugins > Macros > Install... select macro txt file macro gets installed to that menu for the current session
load bead stack
Fiji > File > Open select bead stack file if necessary, split channels (Fiji > Image > Color > Split Channels) if necessary, crop image to a single bead