Flow Analysis of Plasmodium

From: Fred A. Menendez (fmenende@jhsph.edu)
Date: Fri Jan 19 2001 - 11:59:15 EST


Compatriots from the Land of Flow:

A researcher at our institution wants to use flow cytometry to measure the
infection rate of RBCs with malarial parasites by measuring the malarial
DNA content of the RBCs.  He would use PI and possibly AO as the
fluorochromes. He also would like to differentiate at what phase of the
cell cycle the majority of the cells are in (?) by determining the number
of cells in G0/G1 vs G2/M.

While in theory this sounds feasible, and while I have done just such
measurements on tissue and WBCs, I have no experience with the measurement
of parasitic or RBC nucleic acids.  Of  course, with the exception of
reticulocytes (which can be measured by flow using AO) the vast majority of
circulating RBCs contain no nucleic acid. As such, I have a number of
technical question I need to answer before we begin these experiments.

Any thoughts, ideas, help, or references would be greatly appreciated.
Thank you in advance,

Fred Menendez
Johns Hopkins School of Hygiene and Public Health
Flow Cytometry Core Laboratory



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