Batch Analysis of the ATP Content of Bovine Sperm, Oocytes, and Early Embryos Using a Scintillation Counter to Measure the Chemiluminescence Produced by the Luciferin–Luciferase Reaction

Abstract

This report describes an approach to using a commercial luciferin–luciferase kit and a scintillation counter for batch analysis of the ATP content of bovine sperm, oocytes and embryos. After ATP extraction, the luciferin–luciferase assay mixture was added to as many as 48 samples at one time, and the tubes counted once, and then again, in reverse order. The square-root of the geometric mean of the two counts for a tube was directly proportional to the ATP content. The measured ATP content was directly proportional to the number of sperm, oocytes, or blastocysts in a sample. The assay is sensitive enough to measure the ATP content of a single oocyte or blastocyst, or approximately 1000 sperm. The assay does not require the complicated and expensive automated sampling and mixing apparatus that is required for batch analysis using luminometer counting. This approach may significantly simplify the determination of the ATP content of large numbers of samples.