TTC staining was exploited for detecting infarction volume and was conducted as described previously. In breif, rats were deeply anesthetised with 10% chloral hydrate intraperitoneally and sacrificed by decapitation. The brain was rapidly dissected and sectioned into six coronal blocks in an adult rat brain matrix (Kent Scientific Corporation) with an approximate thickness of 2 mm. These sections were stained by incubating them in a solution of 2% TTC (Sigma, St. Louis, MO, USA) for 30 min at 37°C in the dark and then a solution of 4% paraformaldehyde. TTC was dissolved in physiological saline solution. The unstained areas were considered to be the infract areas, as presented in the figures. The total infarction volume for each slice was calculated by including all brain slices. The possible interference of brain edema to infarction volume was corrected by the following standard method, the noninfarcted area of the ipsilateral hemisphere/total noninfarcted area (from both the ipsilateral and contralateral hemispheres) [20 (link)].
Adult rat brain matrix
Manufactured by Kent Scientific
Sourced in United States
The adult rat brain matrix is a tool used to aid in the systematic and consistent sectioning of an adult rat brain. It provides a standardized platform to hold the brain in a fixed position, enabling precise and reproducible slicing for downstream analysis.
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3 protocols using adult rat brain matrix
1
TTC Staining for Infarct Volume Assessment
2
Cerebral Infarction Assessment in Rats
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Rats were anesthetized intraperitoneally with sodium pentobarbital (40 mg/kg), and their brains were removed and frozen at −20°C for 30 min. Tissues of frozen forebrains were dissected and coronal sliced into 2-mm slices in adult rat brain matrix (Kent scientific Corporation) using a rodent brain matrix slicer. These tissue slices were stained with 2% 2,3,7-triphenyltetrazolium chloride (TTC, Sigma, USA) for 20 min in dark conditions at 37°C, then they were soaked and scanned in 4% paraformaldehyde phosphate buffer for 1 h. The degree of cerebral infarction was represented by the ratio between the infarct area and the entire brain area. Unstained (white) area was considered as the infract area whereas normal brain tissues were stained in red. The Image J 1.46R software (NIH, USA) enabled us to assess the cerebral infarction status: infarct area (white)/total area.
3
Quantifying Cerebral Infarction in Rats
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Rats were anesthetized by intraperitoneal injection of sodium pentobarbital (40 mg/kg), brains were removed and frozen at −20°C for 30 minutes. Tissues of frozen forebrains were dissected and coronally sliced into 2 mm slices from adult rat brain matrix (Kent Scientific Corporation, USA) using a rodent brain matrix slicer. These tissue slices were stained with 2% 2, 3, 7-triphenyltetrazolium chloride (TTC, Sigma, USA) at 37°C for 20 minutes under dark conditions, then slices were fixed in 4% paraformaldehyde phosphate buffer for 1 hour. The degree of cerebral infarction was represented by the ratio of infarct area to the area of the entire brain. The infarct brain area was represented by unstained (white) color whereas normal brain tissues were stained with red color. The Image J 1.46R software (NIH, USA) enabled us to determine the cerebral infarction status which was expressed as a ratio: infarct area (white)/total area (red).
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