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Dynamic fluorescence microscopy of cellular uptake of intercalating model drugs by ultrasound-activated microbubbles

BHA Lammertink, R Deckers, M Derieppe, Ine De Cock, Ine Lentacker UGent, G Storm, CTW Moonen and C Bos (2017) MOLECULAR IMAGING AND BIOLOGY. 19(5). p.683-693
abstract
The combination of ultrasound and microbubbles can facilitate cellular uptake of (model) drugs via transient permeabilization of the cell membrane. By using fluorescent molecules, this process can be studied conveniently with confocal fluorescence microscopy. This study aimed to investigate the relation between cellular uptake and fluorescence intensity increase of intercalating model drugs. SYTOX Green, an intercalating fluorescent dye that displays > 500-fold fluorescence enhancement upon binding to nucleic acids, was used as a model drug for ultrasound-induced cellular uptake. SYTOX Green uptake was monitored in high spatiotemporal resolution to qualitatively assess the relation between uptake and fluorescence intensity in individual cells. In addition, the kinetics of fluorescence enhancement were studied as a function of experimental parameters, in particular, laser duty cycle (DC), SYTOX Green concentration and cell line. Ultrasound-induced intracellular SYTOX Green uptake resulted in local fluorescence enhancement, spreading throughout the cell and ultimately accumulating in the nucleus during the 9-min acquisition. The temporal evolution of SYTOX Green fluorescence was substantially influenced by laser duty cycle: continuous laser (100 % DC) induced a 6.4-fold higher photobleaching compared to pulsed laser (3.3 % DC), thus overestimating the fluorescence kinetics. A positive correlation of fluorescence kinetics and SYTOX Green concentration was found, increasing from 0.6 x 10(-3) to 2.2 x 10(-3) s(-1) for 1 and 20 mu M, respectively. Finally, C6 cells displayed a 2.4-fold higher fluorescence rate constant than FaDu cells. These data show that the temporal behavior of intracellular SYTOX Green fluorescence enhancement depends substantially on nuclear accumulation and not just on cellular uptake. In addition, it is strongly influenced by the experimental conditions, such as the laser duty cycle, SYTOX Green concentration, and cell line.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
Model drug, Fluorescence, Confocal microscopy, Ultrasound, Microbubbles, Drug delivery, MEDIATED SONOPORATION, MEMBRANE POROSITY, REAL-TIME, IN-VITRO, DELIVERY, ENDOCYTOSIS, INDUCTION, CELLS, GREEN
journal title
MOLECULAR IMAGING AND BIOLOGY
Mol. Imaging. Biol.
volume
19
issue
5
pages
683 - 693
Web of Science type
Article
Web of Science id
000408699500005
ISSN
1536-1632
1860-2002
DOI
10.1007/s11307-016-1042-x
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
8539486
handle
http://hdl.handle.net/1854/LU-8539486
date created
2017-11-28 13:02:47
date last changed
2017-12-13 09:11:56
@article{8539486,
  abstract     = {The combination of ultrasound and microbubbles can facilitate cellular uptake of (model) drugs via transient permeabilization of the cell membrane. By using fluorescent molecules, this process can be studied conveniently with confocal fluorescence microscopy. This study aimed to investigate the relation between cellular uptake and fluorescence intensity increase of intercalating model drugs.
SYTOX Green, an intercalating fluorescent dye that displays {\textrangle} 500-fold fluorescence enhancement upon binding to nucleic acids, was used as a model drug for ultrasound-induced cellular uptake. SYTOX Green uptake was monitored in high spatiotemporal resolution to qualitatively assess the relation between uptake and fluorescence intensity in individual cells. In addition, the kinetics of fluorescence enhancement were studied as a function of experimental parameters, in particular, laser duty cycle (DC), SYTOX Green concentration and cell line.
Ultrasound-induced intracellular SYTOX Green uptake resulted in local fluorescence enhancement, spreading throughout the cell and ultimately accumulating in the nucleus during the 9-min acquisition. The temporal evolution of SYTOX Green fluorescence was substantially influenced by laser duty cycle: continuous laser (100 \% DC) induced a 6.4-fold higher photobleaching compared to pulsed laser (3.3 \% DC), thus overestimating the fluorescence kinetics. A positive correlation of fluorescence kinetics and SYTOX Green concentration was found, increasing from 0.6 x 10(-3) to 2.2 x 10(-3) s(-1) for 1 and 20 mu M, respectively. Finally, C6 cells displayed a 2.4-fold higher fluorescence rate constant than FaDu cells.
These data show that the temporal behavior of intracellular SYTOX Green fluorescence enhancement depends substantially on nuclear accumulation and not just on cellular uptake. In addition, it is strongly influenced by the experimental conditions, such as the laser duty cycle, SYTOX Green concentration, and cell line.},
  author       = {Lammertink, BHA and Deckers, R and Derieppe, M and De Cock, Ine and Lentacker, Ine and Storm, G and Moonen, CTW and Bos, C},
  issn         = {1536-1632},
  journal      = {MOLECULAR IMAGING AND BIOLOGY},
  keyword      = {Model drug,Fluorescence,Confocal microscopy,Ultrasound,Microbubbles,Drug delivery,MEDIATED SONOPORATION,MEMBRANE POROSITY,REAL-TIME,IN-VITRO,DELIVERY,ENDOCYTOSIS,INDUCTION,CELLS,GREEN},
  language     = {eng},
  number       = {5},
  pages        = {683--693},
  title        = {Dynamic fluorescence microscopy of cellular uptake of intercalating model drugs by ultrasound-activated microbubbles},
  url          = {http://dx.doi.org/10.1007/s11307-016-1042-x},
  volume       = {19},
  year         = {2017},
}

Chicago
Lammertink, BHA, R Deckers, M Derieppe, Ine De Cock, Ine Lentacker, G Storm, CTW Moonen, and C Bos. 2017. “Dynamic Fluorescence Microscopy of Cellular Uptake of Intercalating Model Drugs by Ultrasound-activated Microbubbles.” Molecular Imaging and Biology 19 (5): 683–693.
APA
Lammertink, B., Deckers, R., Derieppe, M., De Cock, I., Lentacker, I., Storm, G., Moonen, C., et al. (2017). Dynamic fluorescence microscopy of cellular uptake of intercalating model drugs by ultrasound-activated microbubbles. MOLECULAR IMAGING AND BIOLOGY, 19(5), 683–693.
Vancouver
1.
Lammertink B, Deckers R, Derieppe M, De Cock I, Lentacker I, Storm G, et al. Dynamic fluorescence microscopy of cellular uptake of intercalating model drugs by ultrasound-activated microbubbles. MOLECULAR IMAGING AND BIOLOGY. 2017;19(5):683–93.
MLA
Lammertink, BHA, R Deckers, M Derieppe, et al. “Dynamic Fluorescence Microscopy of Cellular Uptake of Intercalating Model Drugs by Ultrasound-activated Microbubbles.” MOLECULAR IMAGING AND BIOLOGY 19.5 (2017): 683–693. Print.