Sample Preparation for Electron Microscopy

Samples were processed as previously described^{3 (link),24}. In short, samples were osmicated, stained en bloc with uranyl acetate and embedded in EMbed 812, an Epon-812 substitute (EMS). 1 µm sections were cut and stained with toluidine blue and visualized on a light microscope (Leica DM5500B). Additional thin sections were cut, carbon-coated and imaged either on JEOL JEM-1200-EX electron microscope or a T12 electron microscope (FEI).

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Najm F.J., Madhavan M., Zaremba A., Shick E., Karl R.T., Factor D.C., Miller T.E., Nevin Z.S., Kantor C., Sargent A., Quick K.L., Schlatzer D.M., Tang H., Papoian R., Brimacombe K.R., Shen M., Boxer M.B., Jadhav A., Robinson A.P., Podojil J.R., Miller S.D., Miller R.H, & Tesar P.J. (2015). Drug-based modulation of endogenous stem cells promotes functional remyelination in vivo. Nature, 522(7555), 216-220.

Publication 2015

DM5500B JEM-1200-EX electron microscope T12 electron microscope

Carbon Electron microscope Epon 812 Microscope light Thin sections Toluidine blue Uranyl acetate

Corresponding Organization :

Other organizations : Case Western Reserve University, University School, PerkinElmer (United States), University of Cincinnati Medical Center, National Center for Advancing Translational Sciences, National Institutes of Health, Northwestern University

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Variable analysis

independent variables

Sample processing (osmication, staining en bloc with uranyl acetate, embedding in EMbed 812)
Sectioning (1 µm sections, thin sections)

dependent variables

Visualization of 1 µm sections on light microscope (Leica DM5500B)
Imaging of thin sections on electron microscopes (JEOL JEM-1200-EX, T12 electron microscope (FEI))

control variables

Sample preparation and processing (as previously described in reference 3)
Sectioning (1 µm and thin sections)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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