Download Cyto Pulse Sciences, Inc.

PA4000 User Manual: rev.1-1/05
Table of Contents
page
1.
Introduction
1-1
PA-4000 PulseAgile® Systems, Options, and Accessories
2.
Tutorials
2-1
2.1
Electroporation
2-1
2.2
Electroporation Equipment
2-2
2.2.1
2.2.2
2.2.3
2.2.4
2-2
2-4
2-6
2-6
2.3
2.4
2.5
Electric Fields in Aqueous Solutions and "Load"
Exponential Decay (ED) Electroporators
Rectangular Wave Electroporators
Cyto Pulse Sciences PA-4000 PulseAgile® Electroporator
Using and Optimizing PulseAgile® Protocols
2-6
2.3.1
2.3.2
2.3.3
2.3.4
2.3.5
2.3.6
2.3.7
2.3.8
2.3.9
2.3.10
2-7
2-7
2-8
2-8
2-8
2-9
2-9
2-9
2-10
2-10
Background
Initial Pore Formation
Initial Pulse Width
Follow-up High Voltage Pulse Further Pore Formation
Movement of Material into Cells
Cell Viability Factors – Heat
Cell Viability Factors - Excess Voltage
Other Cell Associated Factors
Solution Temperature, Pore Closing Times
Addition of Reagents
Method Development
2-10
2.4.1
2.4.2
2.4.3
2.4.4
2.4.5
2-10
2-11
2-11
2-12
2-12
2-12
2-12
2-13
2-13
2-13
2-13
2.4.6
2.4.7
2.4.8
2.4.9
3.
1-2
Choosing a Starting Point
Electroporation Medium
Reporter Molecules
Cell Viability
Electrical Parameters
2.4.5.1 Published Protocols
2.4.5.2 Cell Diameter
Optimize the first pulse
Optimize multiple, high voltage pulses
Optimize molecular transport
Further optimization of Molecular Transport Pulses
References
2-15
Operational Concepts
3-1
3.1
Important Concepts
3-1
3.1.1
3.1.2
3.1.3
3.1.4
3-1
3-2
3-2
3-4
3-4
3-4
3-6
3-6
3-7
3.1.5
3.1.6
3.1.7
Load Resistance and Conductance
Power Supply Voltage Setting and Voltage Monitor
Relationship Between Power Supply and Pulse Amplitude
Changing Amplitude from Pulse-to-Pulse
3.1.4.1 Decreasing Voltage from One Pulse to the Next
3.1.4.2 Increasing Voltage from One Pulse to the Next
Pre-Pulse Load Estimator
Pulse Droop
Aqueous Solution Heating
Cyto Pulse Sciences, Inc. P. O. Box 609, Columbia, MD 21045 USA 410-715-0990
iii