Microsample preparation by dielectrophoresis: isolation of malaria.
An important enabling factor for realising integrated micro fluidic analysis instruments for medical diagnostics purposes is front-end sample preparation. Dielectrophoresis is a method that offers great potential for cell discrimination and isolation for sample processing, and here we have applied it to the problem of isolating malaria-infected cells from blood. During development of the malarial pathogen, Plasmodium falciparum, increases occur in the ionic permeability of the plasma membrane of infected erythrocytes. When challenged by suspension in a low conductivity medium, infected cells lose internal ions while uninfected cells retain them. The resultant dielectric differences between infected and uninfected cells were exploited by dielectrophoretic manipulation in spatially inhomogeneous, travelling electrical fields produced by two types of microelectrode arrays. Parasitised cells of ring form or later stage from cultures and clinical specimens were isolated by steric dielectric field-flow-fractionation, focused at the centre of a spiral electrode array, and identified and counted. The dielectrophoretic methods require only a few micro litres of blood, and should be applicable to the production of small, low-cost automated devices for assessing parasite concentrations with potential applicability to drug sensitivity studies and the diagnosis of malaria. By simple adjustment of the electrical field parameters, other cell subpopulations that characterise disease, such as residual cancer cells in blood, can be similarly isolated and analysed.
Gascoyne
Peter
P
Department of Molecular Pathology, University of Texas, M D Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77030, USA. peter@dielectrophoresis.org
Mahidol
Chulabhorn
C
Ruchirawat
Mahidol
M
Satayavivad
Jutamaad
J
Watcharasit
Piyajit
P
Becker
Frederick  F
F
D000818:Animals; D004586:Electrophoresis / Q000379:methods*; D004912:Erythrocytes / Q000469:parasitology*; D008839:Microelectrodes; D010963:Plasmodium falciparum / Q000302:isolation & purification*