Loss of viable neurons was determined by Nissl (cresyl violet) staining, and thus it reveals the structural features of neurons. The spinal cord sections described above were used to stain with Nissl as previously described (Sakakima et al., 2012). Cells that contained Nissl substance were considered to be viable neurons. Condensed fragmented staining indicates neuronal degeneration. The viable neurons with visible nucleoli were manually counted using a BX-60 microscope equipped with a DP70 camera unit (Olympus).
Dp70 camera unit
Manufactured by Olympus
Sourced in Japan
The DP70 camera unit is a high-quality digital camera designed for use with Olympus microscopes. It features a 12.5-megapixel CCD sensor and can capture images with a resolution of 4080 x 3072 pixels. The DP70 provides accurate color reproduction and is capable of live video streaming at up to 20 frames per second. It is compatible with a variety of Olympus microscope models and can be integrated into various imaging software applications.
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5 protocols using dp70 camera unit
1
Nissl Staining for Neuronal Viability
2
Spinal Cord Histopathology Analysis
3
Electrochemical Analysis and Microscopic Characterization
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The electrochemical experiments were performed using CHI900B and 750E potentiostats (CH Instruments, Austin, TX) with a three-electrode cell placed in a Faraday cage. A 1 mm diameter Pt wire was used as a counter electrode, and the reference electrode was Ag/AgCl (3 M KCl). Scanning electron microscopy (SEM) was performed using a Sirion FEI XL FEG/SFEG microscope (FEI Co., The Netherlands) with an accelerating voltage of 2 kV. Fluorescence microscopy images were taken using an Olympus IX71 inverted microscope with a mercury lamp and a CCD camera system (DP70 camera unit and PCI board). Dynamic laser scattering (DLS) data was obtained by Zetasizer Nano ZS (Malvern, Westborough, MA). Nanoparticle tracking analysis (NTA) data was obtained by NanoSight NS300 (Malvern, Westborough, MA). 2D and 3D simulation were performed using Comsol Multiphysics.
4
Histological Analysis of EAE Mouse Model
5
Spinal Cord Tissue Preparation for Histology
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Mice were anesthetized and transcardially perfused first with phosphate-buffered saline (PBS) and then 4% paraformaldehyde in PBS as described previously [38 (link)]. The spinal cords were decalcified prior to histological examination. Cryosections (8 μm thick) obtained from the spinal cord lumbar area were used for Hematoxylin and Eosin (H&E) staining or immunostaining for B220 (Invitrogen, Carlsbad, CA USA; cat. no. 14045285). All digital images were taken using a BX-60 microscope equipped with a DP70 camera unit (Olympus, Tokyo, Japan).
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