Measurement of adenosine triphosphate content in single red blood cells using the firefly bioluminescent reaction

Abstract

A unique optical instrument is described which uses the firefly bioluminescent reaction to measure adenosine triphosphate (ATP) levels in single red blood cells. The method allows chemical content level to be associated with individual cell features. The optical instrument consists of a phase contrast microscope to view cells, a pulsed argon-ion laser to rupture the cell membrane, and a photon counting system to measure the bioluminescent yield. The technique has been calibrated against a standard ATP measurement using bulk analysis methods. The ATP loss mechanism for blood cells in a controlled depletion experiment was also investigated.

''Aged'' (dense) circulating red cells contain normal concentrations of ATP

“…The essence of the technique is suspension of red cells in autologous plasma containing luciferin and luciferase, lysis of the cells with a pulse from a laser, and counting of the photoemissions resulting from reaction of the released ATP with the luciferase. Details of the technique are to be published elsewhere (2); preliminary accounts of the technique have been • published (3,4).…”