A MICROSCOPE PERFUSION RESPIROMETER FOR CONTINUOUS RESPIRATION MEASUREMENT OF CULTURED-CELLS DURING MICROSCOPIC OBSERVATION

  • 1 January 1983
    • journal article
    • research article
    • Vol. 87  (1) , 19-34
Abstract
The construction of a microscope perfusion respirometer (MPR) for simultaneous recording of cellular respiration and microscopic morphology is described. All light microscope techniques for living cells (e.g., phase contrast, differential interference contrast (DIC), fluorimetry) can be applied to the monolayer cells grown on a coverslip. The main constituents of the MPR are a precisely operating perfusion pump (constant volume output), a modified Dvorak-Stoner perfusion chamber, a special holder for the Clark-type O2 probe, gas-tight connections of stainless steel tubing with dead volume-free fittings and a temperature control unit. The cell material, established XTH (Xenopus laevis tadpole heart) cells, is characterized. Examples of operation are presented, concerning normal respiration, respiration during uncoupling of oxidative phosphorylation by carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) and lactate production under anoxia. The corresponding mitochondrial in situ morphology is demonstrated on photomicrographs. Details of construction and application are discussed. This new technique may extend the use of cell cultures instead of animal experiments in pharmaceutical routine tests.