For each animal, two coronal tissue sections that included the primary somatosensory cortex (S1) and CA1 field of the hippocampus (HPC) were imaged with a Nikon A1R HD confocal microscope using three excitation wavelengths: 488 (filter cube = 450/50), 561 (filter cube = 525/50), and 640 (filter cube = 595/50) nm simultaneously, using a 20x (Plan Apo VC 20x DIC N2, MRD70200) or 40x water immersion (Apo LWD 40x WI λS DIC N2, MRD77410) objective. All image analysis was carried out offline in NIH ImageJ (RRID:SCR_003070) or FIJI (RRID:SCR_002285) and performed masked to treatment by a single observer per experiment. For each type of analysis, one image per two sections per animal was analyzed. All image analysis protocols were generated in-house and results from identical or similar protocols have been previously published (Lowery et al., 2017 (link); Sipe et al., 2016 (link)).
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Multimodal Imaging of Cortical and Hippocampal Structures
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Corresponding Organization : University of Rochester
Other organizations : University of Arkansas for Medical Sciences
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Variable analysis
independent variables
- Excitation wavelengths (488 nm, 561 nm, 640 nm)
- Fluorescence intensity measured in the primary somatosensory cortex (S1) and CA1 field of the hippocampus (HPC)
- Tissue sections from the same animals
- Imaging with a Nikon A1R HD confocal microscope
- Use of 20x or 40x water immersion objective
- Image analysis performed by a single observer masked to treatment
- Analysis protocols previously published and validated
- Positive controls: Not explicitly mentioned
- Negative controls: Not explicitly mentioned
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