ASC Field Illumination
From BioDIP
(Difference between revisions)
| Line 1: | Line 1: | ||
| − | The two images acquired | + | The two images acquired via [[ASC Fluorescence and Brightfield Imaging|Fluorescence and Brightfield Imaging]] already gave an impression of the field illumination. Homogeneous illumination is particularly important for quantitative analysis, i.e. the comparison of intensities in different regions of the field of view. Therefore the following test provides a more quantitative analysis of the field illumination. |
== requirements == | == requirements == | ||
* mirror slide | * mirror slide | ||
Revision as of 15:53, 30 September 2009
The two images acquired via Fluorescence and Brightfield Imaging already gave an impression of the field illumination. Homogeneous illumination is particularly important for quantitative analysis, i.e. the comparison of intensities in different regions of the field of view. Therefore the following test provides a more quantitative analysis of the field illumination.
requirements
- mirror slide
- alternatively: reflective surface (i.e. glass slide, gold-coated surface)
acquisition
- 80/20 mirror, laser of interest (1 line of each laser), pinhole 1 AU
- focus on reflective side of mirror
- try to obtain "japanese flag"
images
Zeiss mirror slide
"Japanese flag", reflected light image of mirror slide, Zeiss LSM 510, 10x/0.45, zoom 0.7, 594 nm laser, HiLo LUT in Fiji
Fiji: surface plot of illumination pattern
Fiji: surface plot of illumination pattern, inverted
