An improved automated method to quantitate infarct volume in triphenyltetrazolium stained rat brain sections.
BACKGROUND
The identification of acute neuroprotectants relies heavily on rodent stroke models. It is well know that some of the more common models used can exhibit a relatively high degree of inter animal variability. This necessitates the need to increase the sample size per group and to run concomitant positive and negative control groups with each study in order to increase the consistency and reproducibility of the model. As such, one aspect of these studies that has become more labor intensive is the measurement of infarct volume post study.


METHODS
Herein, we describe a simple method to determine stroke infarct volume in triphenyltetrazolium (TTC) stained brain sections utilizing an automated set of routines using standard software. The method was first validated by determining the correlation of infarct volumes derived from the manual measurements vs the automated method for the same samples across a wide range of infarcts.


RESULTS
This comparison resulted in a significant correlation (r=0.99) indicating that the automated method was a valid method to assess infarct volume across a wide range in lesion volumes. Next, the automated infarct analysis tool was used to determine the effect of (+)-MK801, a well known neuroprotectant, on infarct volume after cerebral ischemia. This study demonstrated a significant reduction in infarct volume in (+)-MK801 treated rats.


CONCLUSIONS
These data demonstrate a simple, accurate automated routine to measure lesion volume in TTC stained sections.
Regan
Hillary K
HK
Department of Stroke and Neurodegeneration, Merck Research Laboratories, West Point, PA 19486, USA.
Detwiler
Theodore J
TJ
Huang
Judy C
JC
Lynch
Joseph J
JJ
Regan
Christopher P
CP
D000818:Animals; D001921:Brain / Q000187:drug effects / Q000473:pathology*; D001923:Brain Chemistry; D020520:Brain Infarction / Q000378:metabolism / Q000473:pathology* / Q000517:prevention & control; D016291:Dizocilpine Maleate / Q000008:administration & dosage / Q000627:therapeutic use; D007091:Image Processing, Computer-Assisted / Q000379:methods; D007274:Injections, Intraperitoneal; D018696:Neuroprotective Agents / Q000008:administration & dosage / Q000627:therapeutic use; D051381:Rats; D015203:Reproducibility of Results; D013194:Staining and Labeling / Q000379:methods; D013778:Tetrazolium Salts / Q000737:chemistry*; D013997:Time Factors
D018696:Neuroprotective Agents; D013778:Tetrazolium Salts; D016291:Dizocilpine Maleate; C009591:triphenyltetrazolium