Slides were imaged on an Olympus FV1200 confocal microscope as previously described for analysis.71 (link) Fifteen confocal z-stack images were captured per treatment at 60× magnification. To examine the extent of GR nuclear colocalization, ImageJ software (National Institutes of Health) was utilized. Briefly, DAPI-positive nuclei were masked, and the percent of total GR-positive signal to DAPI-overlapping was calculated as previously described.34 (link),71 (link) A Zeiss LSM 880 AxioObserver laser scanning confocal microscope was used for representative images. A 63×/1.40 PLAN APO oil objective was 1-μm Z-stacked images with Argon (for FKBP51-positive signal in green) and Red HeNe (for GR-positive signal in red).
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Quantitative Confocal Imaging of GR Nuclear Localization
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Corresponding Organization : USF Health Byrd Alzheimer's Institute
Other organizations : Technical University of Munich
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Variable analysis
independent variables
- Treatment
- Extent of GR nuclear colocalization
- Capturing fifteen confocal z-stack images per treatment at 60× magnification
- Using DAPI-positive nuclei to mask and calculate the percent of total GR-positive signal to DAPI-overlapping
- No positive or negative controls were explicitly mentioned.
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