Flow cytometry protocol to evaluate ionizing radiation effects on P-glycoprotein activity

Abstract

The aim of this work was to establish a protocol to evaluate ionizing radiation effects on P-glycoprotein (P-gp) activity. For this, human peripheral blood samples were irradiated in vitro with different doses and P-gp activity was analyzed for CD4 and CD8 T lymphocytes through rhodamine123-efflux assay by flow cytometry. By simultaneous employment of percentage and mean fluorescence index parameters, subject-by-subject analysis pointed out changes in P-gp activity for some individuals and irradiated samples.Based on this work, the proposed protocol was considered adequate for evaluating P-gp activity on cells after radioactive stress. Besides, this research suggests that P-gp activity could be an important factor to define patient-specific protocols in combined chemo-and radiotherapy, particularly when radiation exposure precedes chemical treatment.

Damage assessment of the equine sperm membranes by fluorimetric technique

“…Among these, great emphasis has been given to fluorescent probes by epifluorescence microscopy or flow cytometry system (Graham et al, 1990Andrade et al, 2007;Celeghini et al, 2008). Fluorescence is a sensible and specific indicator of the state of some molecules, so that it can be applied as a method to measure the metabolic changes in the living cells (Haugland, 2001;Santos et al, 2008). The integrity of sperm membrane is crucial for the maintenance of the sperm fertilizing capability.…”