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PA-3000 User Manual
starting point. With a fixed number of pulses and pulse width, test the effect of changing voltage
through the range of voltages. Next test the range of pulse widths against the optimal voltage.
Repeat this process until an optimum is found.
2
6
(x 10 )
calcein molecules/cell
Again, employing a factorial analysis by modifying electric field, pulse width and pulse
number simultaneously may save time. The reason for the increased efficiency is that with a
factorial design interactions can be assessed and experimental variability is measured over the
entire assay rather than just repetitions at individual independent variables.
1
0
Single Pulse
PulseAgile
Figure: 2.7 Example, Uptake Calcein
Fig 2.7: Molecular uptake of calcein (a fluorescent tracer molecule) is enhanced with
PulseAgile® protocols compared to a single pulse protocol. The single pulse protocol was
applied at 3.3 kV/cm for 50 µs. The PulseAgile® protocols included a single pulse (3.3 kV/cm,
50 µs) followed 0.125 seconds later by 10 pulses of 1 ms and 0.4 kV/cm with interval of either
(„) 0.125 s or (░) 20 sec. DU 145 prostate cancer cells were used at 2 X 106 cells/ml in a 2 mm
gap cuvette and 10 µM calcein. Molecular transport and cell viability were calculated using
calibrated flow cytometry with propidium iodide as the viability stain. (Mark Prausnitz, Ph.D.,
Georgia Institute of Technology provided data for this and the following graph)
cell viability (%)
1 00
50
0
S in g le P u lse
P u lse A g ile
Figure: 2.8 Viability
Fig 2.8: Cell viability was shown not to decrease with the PulseAgile® protocols. The
pulse protocols used were the same as those used in Fig. 2.7.
Cyto Pulse Sciences, Inc., P. O. Box 609, Columbia, MD 21045, Voice: 410-715-0990
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