Mitochondrial DNA Damage Assessment Using Fluorescence Microscopy Quantitation

Abstract

Current methods of fluorescence quantitation are often costly, technically complex, and prone to error. A relatively efficient and cost effective method of fluorescence quantitation is described, which results in quantifiable data. Photographic images of control and test sections are produced under the same conditions of image capture, processing, and print production. The resultant contact sheet is scanned into the NIH Image 1.6 program on a Macintosh computer. Selected areas of these images are measured, resulting in fluorescence mean and standard deviation data that are statistically analyzed. The method was validated by comparison with 3 other methods of fluorescence quantitation: analysis of digitized images, confocal laser scanning microscopy, and densitometry of photographic negatives. Analysis of positives, and to a lesser extent densitometry of negatives, proved to be faster, more efficient, less error prone and cost effective alternatives to the more established methods. (The J Histotechnol 22:17, 1999)

Keywords:

• densectometry
• digitization
• image analysis
• laser scanning