Comet Assay Analysis
Oxidative DNA damage is evaluated using the Comet assay. To quantitate the Comet assay, prepared samples are examined under a Nikon fluorescence microscope. Fifty to one hundred comets per slide are visually scored according to the amount of DNA present in the tail. The Olive tail moment = [(fluorescence intensity of the tail)/(fluorescence intensity of the head) x tail length], is used as a measure of DNA damage. (Klaunig JE)
The Tecan Infinite M200 Microplate Reader is used to determine the amount of anti-oxidant in the sample. Magellan6 software converts instrument-generated WSP files to Excel spreadsheet format. Values are compared to standard curves. (Klaunig JE)
Fluorescence Microscopy – Nuclear Activity Assays
Fluorescence microscopy is used to analyze nuclear activity assays.
“Cell culture involves the dispersal of cells in an artificial environment composed of nutrient solutions, a suitable surface to support the growth of cells, and ideal conditions of temperature, humidity, and gaseous atmosphere. In such a system, a researcher can precisely measure the response of the cell’s alterations in culture alterations, prospective drugs, the presence or absence of other kinds of cells, carcinogenic agents, and viruses.” (Coriell Institue for Cellular Research)
“qPCR and, more specifically, real-time qPCR has become a routine and robust approach for measuring the expression of genes of interest, validating microarray experiments, and monitoring biomarkers.” (Biotechniques)