Upright axio imager z1 microscope

Brightfield and wide-field fluorescence imaging was performed with a Zeiss upright Axio-Imager Z1 microscope fitted with Axio- Cam HRc and HRm cameras and Apotome. Images were acquired with AxioVision software (Carl Zeiss) or ZEN blue (Carl Zeiss). For IF, matched sections from 6 brains were imaged per condition using identical acquisition settings to allow for quantification. Following this, matched neocortical regions of 100 μm width were cropped from each section in Photoshop (Adobe) and imported into ImageJ software (NIH). To generate fluorescence intensity histograms, cropped sections were divided into 20 bins of equal size across the ventricular to pial surfaces and fluorescence intensity corresponding to NFIA or NFIB was normalized to DAPI intensity within each bin. Statistical significance was determined using one-way ANOVA, with p-values below 0.05 considered significant. All values are presented as the mean, with error bars representing the standard error of the mean. Images in figures are representative images that have been identically cropped, enhanced for contrast and brightness, and pseudo-colored to permit overlay using Adobe Photoshop software.

Brightfield imaging was performed with a Zeiss upright Axio-Imager Z1 microscope fitted with an Axio-Cam HRc camera. Confocal fluorescent images were acquired as single 0.6-μm-thick optical sections using a Zeiss inverted Axio-Observer fitted with a W1 Yokogawa spinning disk module and Hamamatsu Flash4.0 sCMOS camera and Slidebook 5.5 software. Images were pseudocoloured to permit overlay and then were cropped, sized and contrast-brightness enhanced for presentation with Adobe Photoshop software.