Testing for electrotransfection parameters by use of the fluorescent dye Lucifer Yellow CH

Georg Sczakiel; Rainer Döffinger; MICHAEL PAWLITA

doi:10.1016/0003-2697(89)90248-0

Testing for electrotransfection parameters by use of the fluorescent dye Lucifer Yellow CH

Georg Sczakiel^*, Rainer Döffinger, MICHAEL PAWLITA

^*Corresponding author for this work

Institute of Molecular Medicine

German Cancer Research Center

6 Citations (Scopus)

Abstract

A nonradioactive and rapid method to systematically optimize conditions for electrotransfection is described here for a critical parameter, the initial voltage. This technique utilizes the electric field-dependent transfer of the fluorescent compound Lucifer Yellow CH into cells. Dye uptake can be followed and quantified by fluorescence microscopy for individual cells or in sum by fluorescence spectroscopy. Electrotransfection conditions for maximal dye and DNA uptake correspond with each other. Cotransfection of Lucifer Yellow CH and DNA coding for the indicator gene chloramphenicol acetyltransferase followed by fluorescence-activated cell sorting demonstrates that the cell population that takes up the fluorescent compound also expresses the indicator gene.

Original language	English
Journal	Analytical Biochemistry
Volume	181
Issue number	2
Pages (from-to)	309-314
Number of pages	6
ISSN	0003-2697
DOIs	https://doi.org/10.1016/0003-2697(89)90248-0
Publication status	Published - 09.1989

Funding

This work was supported in part by BMFT Grant FKZ 11-083-89. We thank L. Mir for initial suggestions, R. S. Goody for helpful advice with fluorescence !spectroscopy, M. Stoehr for the cell sorting, and H. G&zmann for technical assistance. We also thank H. zur Hausen for his continued encouragement and support.

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title = "Testing for electrotransfection parameters by use of the fluorescent dye Lucifer Yellow CH",

abstract = "A nonradioactive and rapid method to systematically optimize conditions for electrotransfection is described here for a critical parameter, the initial voltage. This technique utilizes the electric field-dependent transfer of the fluorescent compound Lucifer Yellow CH into cells. Dye uptake can be followed and quantified by fluorescence microscopy for individual cells or in sum by fluorescence spectroscopy. Electrotransfection conditions for maximal dye and DNA uptake correspond with each other. Cotransfection of Lucifer Yellow CH and DNA coding for the indicator gene chloramphenicol acetyltransferase followed by fluorescence-activated cell sorting demonstrates that the cell population that takes up the fluorescent compound also expresses the indicator gene.",

author = "Georg Sczakiel and Rainer D{\"o}ffinger and MICHAEL PAWLITA",

note = "Funding Information: This work was supported in part by BMFT Grant FKZ 11-083-89. We thank L. Mir for initial suggestions, R. S. Goody for helpful advice with fluorescence !spectroscopy, M. Stoehr for the cell sorting, and H. G\&zmann for technical assistance. We also thank H. zur Hausen for his continued encouragement and support.",

year = "1989",

month = sep,

doi = "10.1016/0003-2697(89)90248-0",

language = "English",

volume = "181",

pages = "309--314",

journal = "Analytical Biochemistry",

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T1 - Testing for electrotransfection parameters by use of the fluorescent dye Lucifer Yellow CH

AU - Sczakiel, Georg

AU - Döffinger, Rainer

AU - PAWLITA, MICHAEL

N1 - Funding Information: This work was supported in part by BMFT Grant FKZ 11-083-89. We thank L. Mir for initial suggestions, R. S. Goody for helpful advice with fluorescence !spectroscopy, M. Stoehr for the cell sorting, and H. G&zmann for technical assistance. We also thank H. zur Hausen for his continued encouragement and support.

PY - 1989/9

Y1 - 1989/9

N2 - A nonradioactive and rapid method to systematically optimize conditions for electrotransfection is described here for a critical parameter, the initial voltage. This technique utilizes the electric field-dependent transfer of the fluorescent compound Lucifer Yellow CH into cells. Dye uptake can be followed and quantified by fluorescence microscopy for individual cells or in sum by fluorescence spectroscopy. Electrotransfection conditions for maximal dye and DNA uptake correspond with each other. Cotransfection of Lucifer Yellow CH and DNA coding for the indicator gene chloramphenicol acetyltransferase followed by fluorescence-activated cell sorting demonstrates that the cell population that takes up the fluorescent compound also expresses the indicator gene.

AB - A nonradioactive and rapid method to systematically optimize conditions for electrotransfection is described here for a critical parameter, the initial voltage. This technique utilizes the electric field-dependent transfer of the fluorescent compound Lucifer Yellow CH into cells. Dye uptake can be followed and quantified by fluorescence microscopy for individual cells or in sum by fluorescence spectroscopy. Electrotransfection conditions for maximal dye and DNA uptake correspond with each other. Cotransfection of Lucifer Yellow CH and DNA coding for the indicator gene chloramphenicol acetyltransferase followed by fluorescence-activated cell sorting demonstrates that the cell population that takes up the fluorescent compound also expresses the indicator gene.

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DO - 10.1016/0003-2697(89)90248-0

M3 - Journal articles

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EP - 314

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 2

ER -

Testing for electrotransfection parameters by use of the fluorescent dye Lucifer Yellow CH

Abstract

Funding

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