Author: Evonuk, Kirsten S.; Moseley, Carson E.; Doyle, Ryan E.; Weaver, Casey T.; DeSilva, Tara M.
Title: Determining Immune System Suppression versus CNS Protection for Pharmacological Interventions in Autoimmune Demyelination Document date: 2016_9_12
ID: vr83284f_4
Snippet: 1. Tissue preparation 1. Sacrifice EAE mice in a separate experiment from those used in step 3 and its sub-steps at any point after EAE induction (often ~ day 30, during the chronic phase of disease for C57Bl/6 mice or during a peak in average clinical scores for SJL mice) following the steps below to determine extent of reactive gliosis and demyelination. 2. Anesthetize mice with 2.5% isoflurane and 97.5% oxygen and confirm appropriate depth of .....
Document: 1. Tissue preparation 1. Sacrifice EAE mice in a separate experiment from those used in step 3 and its sub-steps at any point after EAE induction (often ~ day 30, during the chronic phase of disease for C57Bl/6 mice or during a peak in average clinical scores for SJL mice) following the steps below to determine extent of reactive gliosis and demyelination. 2. Anesthetize mice with 2.5% isoflurane and 97.5% oxygen and confirm appropriate depth of anesthesia with a gentle toe pinch using forceps, looking for a lack of response. Perform transcardiac perfusion as described in step 3. 8. When ready, section tissue at 16 μm with a cryostat and mount on electrostatically charged slides. Put every 10 th section on a slide each for brain and spinal cord (for example, slide 1 will have sections 1, 11, and 21, and slide 2 will have sections 2, 12, and 22, and so on). Store slides at -80 o C or use right away for staining.
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