Electroporation Assay - Applied BioPhysics, Inc.
Electroporation Assay - Applied BioPhysics, Inc.
Electroporation Assay - Applied BioPhysics, Inc.
Create successful ePaper yourself
Turn your PDF publications into a flip-book with our unique Google optimized e-Paper software.
Elevated Field Applications<br />
1 <strong>Electroporation</strong><br />
2 Wound healing assay
NORMAL MODE 1 MICROAMP, 10 MILLIVOLTS<br />
ELEVATED FIELD 1 MILLIAMP, A FEW VOLTS<br />
Elevated<br />
current<br />
applied<br />
~200msec<br />
pore formation
Variation of the pulse duration: Lucifer yellow uptake<br />
MDCK Type II cells<br />
Pulse:<br />
4.0 V<br />
40 kHz<br />
50 msec<br />
100 msec<br />
200 msec<br />
500 msec
Uptake of dyes with different molecular weight<br />
Pulse:<br />
40 kHz, 4.0 V, 200 msec<br />
Lucifer Yellow<br />
M = 0.5 kDa<br />
TRITC-dextran<br />
M = 76 kDa<br />
FITC-dextran<br />
M = 250 kDa<br />
Albany Medical College (F. Minnear) has<br />
demonstrated introduction of DNA constructs using<br />
the method and obtained expression of GFP
Monitoring Changes in Cell Shape: Intracellular Agonists<br />
Membrane Permeable Compounds<br />
CPT-cAMP: membrane permeable analogue of cAMP<br />
1,4<br />
1,3<br />
|Z|(t) / |Z|(0)<br />
1,2<br />
1,1<br />
1,0<br />
0,9<br />
0 µM<br />
30 µM<br />
10 µM<br />
3 µM<br />
0.3 µM 1 µM<br />
0,8<br />
0 2 4 6 8 10<br />
t [hrs]<br />
300 µM<br />
100 µM
Monitoring Changes in Cell Shape: Intracellular Agonists<br />
In situ <strong>Electroporation</strong>: Effector Uptake<br />
8-OH-cAMP: membrane-impermeant cAMP-analogue<br />
1: Addition of 8-OH-cAMP<br />
2: <strong>Electroporation</strong><br />
|Z| [kΩ]<br />
12<br />
11<br />
10<br />
9<br />
1<br />
High field<br />
2<br />
100 µM<br />
0 µM<br />
8<br />
4 kHz<br />
7<br />
0 1 2 3 4 5<br />
t [hrs]
www.biophysics.com
Change language