Brain sections from saline-, glutamate- or glutamate-plus-osmotin-treated rat pups (n=5 rats per group) were analyzed after 12 h of drug treatment. Transcardial perfusion was performed with 1X phosphate-buffered saline (PBS) followed by 4% ice-cold paraformaldehyde. The brain tissues were post-fixed overnight in 4% paraformaldehyde and subsequently transferred to 20% sucrose until they sank to the bottom of the tube. The brains were frozen in OCT (Tissue-Tek O.C.T. compound medium, Sakura Finetek USA, Inc., Torrance, CA, USA) and then sectioned into 14 μm sections in the coronal plane with a CM 3050S cryostat (Leica, Wetzlar, Germany). The sections were thaw-mounted on Probe-On positively charged slides (Thermo Fisher Scientific Inc., Waltham, MA, USA).
Probe on positively charged slides
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Probe-On positively charged slides are laboratory equipment used for DNA and RNA immobilization. The slides are coated with a positively charged surface that facilitates the attachment of nucleic acid probes, enabling efficient hybridization during various molecular biology applications.
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3 protocols using probe on positively charged slides
1
Rat Brain Sectioning and Analysis
2
Brain Tissue Preparation for Histology
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As previously described [67 (link)], the mice were subjected to transcardial perfusion with ice-cold PBS followed by 4% neutrally buffered paraformaldehyde (NBP). After post fixing the brain in 4% NBP for 72 h, it was washed with PBS and then transferred to 20% sucrose solution for a further 48 h. The brain tissue was frozen in optimal cutting temperature (OCT) (Tissue-Teks O.C.T. Compound Medium, Sakura Finetek USA, Inc., Torrance, CA, USA) and sectioned into 14–16 μm sections in the coronal plane with a CM 3050S cryostat (Leica, Wetzlar, Germany). The sections were thaw mounted on Probe-On positively charged slides (Thermo Fisher Scientific Inc., Waltham, MA, USA).
3
Morphological Brain Tissue Analysis
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The animals were euthanized after 12 h of drug treatment to conduct morphological studies as we reported earlier [26 (link)]. Briefly, the brain tissues from all the treated groups after 12 h were subjected to transcardial perfusion with 4 % ice-cold paraformaldehyde. After postfixing these brain tissues, 4 % paraformaldehyde was transferred to 20 % sucrose. The tissues were frozen in OCT (Tissue-Tek O.C.T. Compound Medium, Sakura Finetek USA, Inc., Torrance, CA, USA), sectioned into 14–16-μm sections in the coronal plane with a CM 3050S cryostat (Leica, Wetzlar, Germany). The sections were thaw-mounted on Probe-On positively charged slides (Thermo Fisher Scientific Inc., Waltham, MA, USA).
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