Advanced system check
From BioDIP
(Difference between revisions)
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Advanced System Check | Advanced System Check | ||
− | Protocol (draft) | + | == Protocol (draft) == |
[[image:Conva10xok.jpg with bar.jpg|thumb]] | [[image:Conva10xok.jpg with bar.jpg|thumb]] | ||
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**requirements: 0.2/0.5µm fluorescent beads sample | **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) | **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 |
Revision as of 16:00, 28 September 2009
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