Eppendorf tubes such that the bacteria concentration is different in each tube. Measure the OD 600 and emission fluorescence intensity of YOPRO at 509 nm of each solution. Plot the
fluorescence intensity as a function of the corresponding number of fluorescent bacteria. Because all the bacteria are lysed,
they all uptake YOPRO, and the fluorescence and OD 600
signals can be related to the number of bacteria containing
YOPRO.
19. The number of irreversibly electroporated bacteria can be calculated from the emission fluorescence intensity of PI using a
calibration curve. This calibration curve is obtained similarly to
the YOPRO calibration, but using 3 μM PI. The fluorescence
intensity of each solution at 618 nm is related to the
corresponding number of fluorescent bacteria using the
OD 600 .
Acknowledgments
This work was supported by the University of Florida.
References
1. Schoenbach KH, Peterkin FE, Alden RW,
Beebe SJ (1997) The effect of pulsed electric
fields on biological cells: experiments and
applications. IEEE Trans Plasma Sci 25
(2):284–292. https://doi.org/10.1109/27.
602501
2. Kanduser M, Miklavcic D (2008) Electroporation in biological cell and tissue: an overview.
In: Vorobiev E, Lebovka N (eds) Electrotechnologies for extraction from food plants
and biomaterials, Food Engineering Series,
vol 233. Springer, New York, NY, pp 1–37
3. Weaver JC, Chizmadzhev YA (1996) Theory of
electroporation: a review. Bioelectrochem
Bioenerg 41(2):135–160. https://doi.org/
10.1016/s0302-4598(96)05062-3
4. Kotnik T, Frey W, Sack M, Meglic SH,
Peterka
M,
Miklavcic
D
(2015)
Electroporation-based applications in biotechnology. Trends Biotechnol 33(8):480–488.
https://doi.org/10.1016/j.tibtech.2015.06.
002
5. Gehl J (2003) Electroporation: theory and
methods, perspectives for drug delivery, gene
therapy and research. Acta Physiol Scand 177
(4):437–447.
https://doi.org/10.1046/j.
1365-201X.2003.01093.x
6. Luo D, Saltzman WM (2000) Synthetic DNA
delivery systems. Nat Biotechnol 18(1):33–37
7. Garcia PA, Ge ZF, Moran JL, Buie CR (2016)
Microfluidic screening of electric fields for electroporation. Sci Rep 6:11. https://doi.org/10.
1038/srep21238
8. Movahed S, Li DQ (2011) Microfluidics cell
electroporation. Microfluid Nanofluid 10
(4):703–734.
https://doi.org/10.1007/
s10404-010-0716-y
9. Lee WG, Demirci U, Khademhosseini A
(2009) Microscale electroporation: challenges
and perspectives for clinical applications. Integr
Biol
1(3):242–251.
https://doi.org/10.
1039/b819201d
10. Geng T, Lu C (2013) Microfluidic electroporation for cellular analysis and delivery. Lab
Chip 13(19):3803–3821. https://doi.org/
10.1039/c3lc50566a
11. Experton J, Wilson AG, Martin CR (2016)
Low-voltage flow-through electroporation in
gold-microtube membranes. Anal Chem 88
(24):12445–12452.
https://doi.org/10.
1021/acs.analchem.6b03820
12. Experton J, Martin CR (2018) The effect of
voltage charging on the transport properties of
gold nanotube membranes. Small 0
54
Juliette Experton et al.
fluorescence intensity as a function of the corresponding number of fluorescent bacteria. Because all the bacteria are lysed,
they all uptake YOPRO, and the fluorescence and OD 600
signals can be related to the number of bacteria containing
YOPRO.
19. The number of irreversibly electroporated bacteria can be calculated from the emission fluorescence intensity of PI using a
calibration curve. This calibration curve is obtained similarly to
the YOPRO calibration, but using 3 μM PI. The fluorescence
intensity of each solution at 618 nm is related to the
corresponding number of fluorescent bacteria using the
OD 600 .
Acknowledgments
This work was supported by the University of Florida.
References
1. Schoenbach KH, Peterkin FE, Alden RW,
Beebe SJ (1997) The effect of pulsed electric
fields on biological cells: experiments and
applications. IEEE Trans Plasma Sci 25
(2):284–292. https://doi.org/10.1109/27.
602501
2. Kanduser M, Miklavcic D (2008) Electroporation in biological cell and tissue: an overview.
In: Vorobiev E, Lebovka N (eds) Electrotechnologies for extraction from food plants
and biomaterials, Food Engineering Series,
vol 233. Springer, New York, NY, pp 1–37
3. Weaver JC, Chizmadzhev YA (1996) Theory of
electroporation: a review. Bioelectrochem
Bioenerg 41(2):135–160. https://doi.org/
10.1016/s0302-4598(96)05062-3
4. Kotnik T, Frey W, Sack M, Meglic SH,
Peterka
M,
Miklavcic
D
(2015)
Electroporation-based applications in biotechnology. Trends Biotechnol 33(8):480–488.
https://doi.org/10.1016/j.tibtech.2015.06.
002
5. Gehl J (2003) Electroporation: theory and
methods, perspectives for drug delivery, gene
therapy and research. Acta Physiol Scand 177
(4):437–447.
https://doi.org/10.1046/j.
1365-201X.2003.01093.x
6. Luo D, Saltzman WM (2000) Synthetic DNA
delivery systems. Nat Biotechnol 18(1):33–37
7. Garcia PA, Ge ZF, Moran JL, Buie CR (2016)
Microfluidic screening of electric fields for electroporation. Sci Rep 6:11. https://doi.org/10.
1038/srep21238
8. Movahed S, Li DQ (2011) Microfluidics cell
electroporation. Microfluid Nanofluid 10
(4):703–734.
https://doi.org/10.1007/
s10404-010-0716-y
9. Lee WG, Demirci U, Khademhosseini A
(2009) Microscale electroporation: challenges
and perspectives for clinical applications. Integr
Biol
1(3):242–251.
https://doi.org/10.
1039/b819201d
10. Geng T, Lu C (2013) Microfluidic electroporation for cellular analysis and delivery. Lab
Chip 13(19):3803–3821. https://doi.org/
10.1039/c3lc50566a
11. Experton J, Wilson AG, Martin CR (2016)
Low-voltage flow-through electroporation in
gold-microtube membranes. Anal Chem 88
(24):12445–12452.
https://doi.org/10.
1021/acs.analchem.6b03820
12. Experton J, Martin CR (2018) The effect of
voltage charging on the transport properties of
gold nanotube membranes. Small 0
54
Juliette Experton et al.