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Quantification of apoptosis

KN Kevin K. Noguchi
SJ Stephen A. Johnson
FM Francesca M. Manzella
KM Kobe L. Masuoka
SW Sasha L. Williams
LM Lauren D. Martin
GD Gregory A. Dissen
CI Chrysanthy Ikonomidou
KS Katie J. Schenning
JO John W. Olney
AB Ansgar M. Brambrink
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The stereological quantitative analysis of cellular apoptosis was conducted by an investigator blinded to the treatment conditions using serial sections cut at 2 mm intervals across the entire brain, and stained with AC3 to mark apoptotic profiles. Sections were quantified at 10x magnification using a computer-assisted Microbrightfield Stereo-Investigator system to estimate the number of AC3-positive (apoptotic) profiles. It was possible to distinguish apoptotic neurons from apoptotic oligodendrocytes because, as we have illustrated previously3–5,35,50, the AC3 stain displays the full cell body and processes of both cell types, and they are morphologically dissimilar. Moreover, the dying oligodendrocytes are confined almost exclusively to white matter and the dying neurons are confined exclusively to gray matter.

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