Spinning-disk confocal image stacks were taken with a 63x/1.40 NA using a BD CARVII (BD Bioscience) spinning-disk confocal microscope accessory fitted to the Zeiss 200M inverted microscope as described previously [26 (link)]. Widefield images were taken using a 63x/1.40 numerical aperture (NA) objective with an Optovar setting of 1.6x or 2.5x on a Zeiss 200M inverted microscope (Carl Zeiss MicroImaging Inc., Thornwood, NY, USA). Image stacks were deconvolved using 10 iterations, Huygens Professional software (version 09:26:16; Scientific Volume Imaging, Hilversum, the Netherlands). Maximum intensity projections (MIPs) and mean pixel intensity measurements were made with iVision-Mac™ software (version 4.5.5; BioVision Technologies). Super resolution (3D SIM) stacks were taken using a Deltavision OMX™ super resolution imaging microscope (GE Healthcare) with an Olympus 60x/NA 1.42 or 100x/NA 1.49 objective and analyzed using FIJI software (version 2.0.0-rc-34/1.50a) and iVision-Mac™ software.
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Spinning-disk Confocal Imaging and Super-resolution
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Corresponding Organization : University of Arkansas for Medical Sciences
Other organizations : National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health
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Variable analysis
independent variables
- None explicitly mentioned
- Maximum intensity projections (MIPs)
- Mean pixel intensity measurements
- Spinning-disk confocal microscope accessory (BD CARVII)
- Zeiss 200M inverted microscope
- 63x/1.40 NA objective
- Optovar setting of 1.6x or 2.5x (for widefield images)
- Deconvolution using 10 iterations, Huygens Professional software
- IVision-Mac™ software for image analysis
- Deltavision OMX™ super resolution imaging microscope
- Olympus 60x/NA 1.42 or 100x/NA 1.49 objective (for super resolution imaging)
- No positive or negative controls were explicitly mentioned in the provided information.
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