After flow cytometric based CMA3 staining assay, samples which were exposed to CMA3 were washed twice with PBS and assessed by Partec
PAS flow cytometer, using an argon laser with an excitation wave length of 488 nm.
The cells were incubated at 37[degrees]C with 5 [micro]L of Annexin V-fluorescein isothiocyanate (FITC) and 10 [micro]L of propidium iodide for 10 min and analyzed with the CyFlow Space (Partec
) and FloMax 2.82 software.
Samples were read using a Partec
Cube 8 flow cytometer with gates set at 2 x [10.sup.5] particles, and data was analyzed using FlowJo.
The blood samples for CD4 estimation were analysed using the Partec
CyFlow machine (Germany).
After washing and resuspending to a hematocrit of 1-2%, the percentage of FITC-labeled cells was measured by flow cytometry using the Partec
PAS-III particle analyzing system (Partec
GMbH) with argon laser (488 nm).
Mudzori, "Evaluation of the partec
flow cytometer against the BD FACSCalibur system for monitoring immune responses of human immunodeficiency virus-infected patients in Zimbabwe," Clinical and Vaccine immunology, vol.
The cells were analyzed using a CyFlowSpace flow cytometer (Partec
MMP was measured via fluorescence-activated cell sorting (FACS; Partec
, Gorlitz, Germany) after staining cells with Rh123, a membrane-permeable fluorescent dye .
The nuclei suspension was analyzed in the Partec
(model PAS II/III; Partec
GmbH; Munster, Germany) flow cytometer.
The nuclear DNA contents of leaves derived from in the cultures were measured with Ploidy Analyser equipment (Partec
GmbH, Germany) using the protocol described by Mishiba et al.
After 20 min incubation at 37[degrees]C, the samples were washed twice, resuspended in 500 [micro]l phosphate-buffered saline (PBS) and analyzed by flow cytometry (Partec
CyFlow[R] Space instrument, Partec
GmbH, Munster, Germany).