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Conference Paper: A study on the change in plasma membrane potential during sonoporation

TitleA study on the change in plasma membrane potential during sonoporation
Authors
Issue Date2013
PublisherISTU2013.
Citation
The 13th International Symposium on Therapeutic Ultrasound (ISTU 2013), Shanghai, China, 12-16 May 2013. In 13th ISTU Final Programme, 2013, p. 152 How to Cite?
AbstractOBJECTIVES: There has been validated that the correlation of sonoporation with calcium transients is generated by ultrasound-mediated microbubbles activity. Besides calcium, other ionic flows are likely involved in sonoporation. Our hypothesis is the cell electrophysiological properties are related to the intracellular delivery by ultrasound and microbubbles. In this study, a real-time live cell imaging platform is used to determine whether plasma membrane potential change is related to the sonoporation process at the cellular level. METHODS: Hela cells were cultured in DMEM supplemented with 10% FBS in Opticell Chamber at 37 °C and 5% CO2, and reached 80% confluency before experiments. The Calcein Blue-AM, DiBAC4(3) loaded cells in the Opticell chamber filled with PI solution and Sonovue microbubbles were immerged in a water tank on a inverted fluorescence microscope. Pulsed ultrasound (1MHz freq., 20 cycles, 20Hz PRF, 0.2-0.5MPa PNP) was irradiated at the angle of 45° to the region of interest for 1s.The real-time fluorescence imaging for different probes was acquired by a cooled CCD camera every 20s for 10min. The time-lapse fluorescence images were quantitatively analyzed to evaluate the correlation of cell viability, intracellular delivery with plasma membrane potential change. RESULTS: Our preliminary data showed that the PI fluorescence, which indicated intracellular delivery, was immediately accumulated in cells adjacent to microbubbles after exposure, suggesting that their membranes were damaged by ultrasound-activated microbubbles. However, the fluorescence reached its highest level within 4 to 6 minutes and was unchanged thereafter, indicating the membrane was gradually repaired within this period. Furthermore, using DIBAC4(3), which detected the change in the cell membrane potential, we found that the loss of membrane potential might be associated with intracellular delivery, because the PI fluorescence accumulation was usually accompanied with the change in DIBAC4 (3) fluorescence. CONCLUSIONS: Our study suggests that there may be a linkage between the cell membrane potential change and intracellular delivery mediated by ultrasound and microbubbles. We also suggest that other ionic flows or ion channels may be involved in the cell membrane potential change in sonoporation. Further efforts to explore the cellular mechanism of this phenomenon will improve our understanding of sonoporation.
DescriptionPosters: no. 4
Control ID: 1680329
Persistent Identifierhttp://hdl.handle.net/10722/189836

 

DC FieldValueLanguage
dc.contributor.authorQin, Pen_US
dc.contributor.authorCai, Pen_US
dc.contributor.authorDu, Len_US
dc.contributor.authorJin, Len_US
dc.contributor.authorYu, Aen_US
dc.date.accessioned2013-09-17T15:00:47Z-
dc.date.available2013-09-17T15:00:47Z-
dc.date.issued2013en_US
dc.identifier.citationThe 13th International Symposium on Therapeutic Ultrasound (ISTU 2013), Shanghai, China, 12-16 May 2013. In 13th ISTU Final Programme, 2013, p. 152en_US
dc.identifier.urihttp://hdl.handle.net/10722/189836-
dc.descriptionPosters: no. 4-
dc.descriptionControl ID: 1680329-
dc.description.abstractOBJECTIVES: There has been validated that the correlation of sonoporation with calcium transients is generated by ultrasound-mediated microbubbles activity. Besides calcium, other ionic flows are likely involved in sonoporation. Our hypothesis is the cell electrophysiological properties are related to the intracellular delivery by ultrasound and microbubbles. In this study, a real-time live cell imaging platform is used to determine whether plasma membrane potential change is related to the sonoporation process at the cellular level. METHODS: Hela cells were cultured in DMEM supplemented with 10% FBS in Opticell Chamber at 37 °C and 5% CO2, and reached 80% confluency before experiments. The Calcein Blue-AM, DiBAC4(3) loaded cells in the Opticell chamber filled with PI solution and Sonovue microbubbles were immerged in a water tank on a inverted fluorescence microscope. Pulsed ultrasound (1MHz freq., 20 cycles, 20Hz PRF, 0.2-0.5MPa PNP) was irradiated at the angle of 45° to the region of interest for 1s.The real-time fluorescence imaging for different probes was acquired by a cooled CCD camera every 20s for 10min. The time-lapse fluorescence images were quantitatively analyzed to evaluate the correlation of cell viability, intracellular delivery with plasma membrane potential change. RESULTS: Our preliminary data showed that the PI fluorescence, which indicated intracellular delivery, was immediately accumulated in cells adjacent to microbubbles after exposure, suggesting that their membranes were damaged by ultrasound-activated microbubbles. However, the fluorescence reached its highest level within 4 to 6 minutes and was unchanged thereafter, indicating the membrane was gradually repaired within this period. Furthermore, using DIBAC4(3), which detected the change in the cell membrane potential, we found that the loss of membrane potential might be associated with intracellular delivery, because the PI fluorescence accumulation was usually accompanied with the change in DIBAC4 (3) fluorescence. CONCLUSIONS: Our study suggests that there may be a linkage between the cell membrane potential change and intracellular delivery mediated by ultrasound and microbubbles. We also suggest that other ionic flows or ion channels may be involved in the cell membrane potential change in sonoporation. Further efforts to explore the cellular mechanism of this phenomenon will improve our understanding of sonoporation.-
dc.languageengen_US
dc.publisherISTU2013.-
dc.relation.ispartof13th International Symposium on Therapeutic Ultrasound Final Programmeen_US
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.titleA study on the change in plasma membrane potential during sonoporationen_US
dc.typeConference_Paperen_US
dc.identifier.emailQin, P: pqin@hku.hken_US
dc.identifier.emailYu, A: alfred.yu@hku.hken_US
dc.identifier.authorityYu, A=rp00657en_US
dc.description.naturepostprint-
dc.identifier.hkuros222141en_US
dc.identifier.spage152-
dc.identifier.epage152-
dc.publisher.placeChina-

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