For immunofluorescence staining and Fluoro-Gold tracing, multichannel fluorescence images of the brain sections were captured using a CCD camera attached to a Carl Zeiss Axiovision fluorescence microscope (Axiovert 200M), and exported as TIFF image files using Axiovision LE (Carl Zeiss, Germany).
Due to the ubiquitous nature of mitochondrial immunoreactivity in the brain, the immunoreactive elements were captured using a 100× oil lens (Axioplan, Zeiss) and the well demarcated punctate elements were manually counted in two 100× fields (6044 µm/field) for each lesion stage and NWM of every acute multiple sclerosis and WMS case.
To measure brain infarct, images from the stained coronal sections were captured using a digital camera and analyzed with the ImageJ (NIH) image processing software.
To quantify neurodegeneration in the cell body and neuropil, images of the 5 µm brain sections stained with hematoxylin and eosin and were captured using bright-field microscopy.
Stained brain slices were photographed using an Olympus Provis AX70 microscope (Olympus America, Center Valley, PA) and images were captured using QCapture Suite (QImaging, Surrey, BC).
Fluorescence images of whole brain coronal sections were obtained using a SteREO Lumar.V12 fluorescence stereoscope (Carl Zeiss MicroImaging GmbH, Oberkochen Germany), whereas striatal images were captured using a Leica TCS 4D confocal scanning laser microscope (Leica Lasertechnik GmbH).
Depending on treatment, 9 11 brain regions and 4 6 sections for each region were examined using light microscopy (Nikon Eclipse E800 series microscope. Images were captured using a CoolSnapHQ camera system and Metamorph software (using 10Xobjective).
Images were captured using a digital camera.
The images were captured using IVIS® Lumina XR Imaging System.
Images were captured using Spot Software Diagnostic Instruments , Inc; Sterling Heights, MI).
