Brain Tissue Harvesting and Histological Analysis

Animal brains were harvested at either 2 weeks post-surgery or following completion of behavioral testing at 2 months, as previously described.^{16 (link)} The entire brain was sectioned at 50-µm intervals on a sliding microtome (Thermo), and serial sections were mounted, dried, and stained with Cresyl Violet or the following antibodies: BrdU [Rat Mab BU1/75(ICR1), Accurate Chemicals], Olig2 (Mouse Mab from C.D. Stiles, Harvard), GFAP (Rat Mab 2.2B10, Thermo), Iba1 (Rabbit Pab, Wako), CC1 (APC, Mouse Mab OP80, Calbiochem), Nkx2.2 (Mouse Mab 74.5A, Developmental Studies Hybridoma Bank), Collagen 4A (Col4A; Goat Pab, AB769 Millipore), Claudin 5 (Mouse Mab, 35-2500 Invitrogen).

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Nguyen V., Chavali M., Larpthaveesarp A., Kodali S., Gonzalez G., Franklin R.J., Rowitch D.H, & Gonzalez F. (2021). Neuroprotective effects of Sonic hedgehog agonist SAG in a rat model of neonatal stroke. Pediatric Research, 90(6), 1161-1170.

Publication 2021

sliding microtome Rabbit Pab Mouse Mab 74.5A

Animal Antibodies Brain Brdu Claudin 5 Collagen Cresyl violet Gfap Goat Hybridoma Microtome Mouse Olig2 Rabbit Surgery

Corresponding Organization :

Other organizations : University of California, San Francisco, Wellcome/MRC Cambridge Stem Cell Institute, University of Cambridge

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Variable analysis

independent variables

Time of brain harvesting: 2 weeks post-surgery or 2 months after completion of behavioral testing

dependent variables

Histological characteristics of the brain sections, including cell types and protein expression (e.g., BrdU, Olig2, GFAP, Iba1, CC1, Nkx2.2, Collagen 4A, Claudin 5)

control variables

Tissue processing: The entire brain was sectioned at 50-μm intervals on a sliding microtome and stained with various antibodies

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

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