Microfluidic sonicator for real-time disruption of eukaryotic cells and bacterial spores for DNA analysis

Marentis, Theodore C.; Kusler, Brenda; Yaralioglu, G.G.; Liu, Shijun; Hæggström, Edward; Khuri‐Yakub, Butrus T.

doi:10.1016/j.ultrasmedbio.2005.05.005

Cited by 62 publications

(48 citation statements)

References 36 publications

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“…When microbubbles are added to the cell suspension, the cavitation effect is greater than when ultrasound is used alone. The mechanical and physical forces of ultrasound exposure will destroy cells if the energy and exposure time are beyond the tolerance limit of the culture, and these forces can even induce cell lysis, leading to death (19,20). Our data indicate that ultrasound combined with SonoVue has a minimal effect on the viability of PC-3 cells.…”

Section: Discussionmentioning

confidence: 77%

The improvement of liposome-mediated transfection of pEGFP DNA into human prostate cancer cells by combining low-frequency and low-energy ultrasound with microbubbles

Bai

Wu²,

Shen³

et al. 2011

Oncol Rep

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Abstract. The aim of this study was to explore the use of a contrast agent to study the effects of exposure to ultrasound, in combination with microbubbles, on liposome-mediated transfection of genes into human prostate cancer cells. A contrast agent was used to study the effects of ultrasound exposure in combination with microbubbles on liposomes, which transfect genes into human prostate cancer cells. The human prostate cancer cell line PC-3 in suspension was exposed to ultrasound with a 20% duty cycle (i.e., 2 sec 'on' time and 8 sec 'off' time) lasting 5 min, with and without ultrasound contrast agent (SonoVue™) using a digital sonifier at a frequency of 21 kHz and an intensity of 4.6 mW/cm 2 . Immediately after exposure to ultrasound, cell viability and membrane damage were measured. After exposure to ultrasound, the cell suspensions were put into 12-well plates and cultured for 24 h. Fluorescence microscopy and flow cytometry were used to detect pEGFP transfection efficiency. Exposure to ultrasound alone and ultrasound combined with microbubbles resulted in minimal cell death and induced negligible cell membrane damage. Ultrasound combined with microbubbles had a greater effect on cell membrane damage in all groups: the average cell membrane damage was 41.87%, and it was approximately 42-fold greater than in the control group. The average transfection efficiency of PC-3 cells was 20.30% for the liposome (Lipofectamine™)+pEGFP+ultrasound+ultrasound contrast agent (SonoVue) group; this was the highest rate of all groups measured and was approximately 81-fold greater than that of the control group. The use of low-frequency and low-energy ultrasound, in combination with microbubbles, could be a potent physical method for increasing liposome gene delivery efficiency. This technique is a promising non-viral approach that can be used in prostate cancer gene therapy.

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Section: Discussionmentioning

confidence: 77%

The improvement of liposome-mediated transfection of pEGFP DNA into human prostate cancer cells by combining low-frequency and low-energy ultrasound with microbubbles

Bai

Wu²,

Shen³

et al. 2011

Oncol Rep

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“…Effects of ultrasound and microbubbles on HUVEC viability The numbers of viable HUVECs decreased linearly with ultrasound exposure time, as shown in Figure 2A, but the viability was still more than 75% even at the effective exposure time of 12 s. The mechanical and physical forces of the ultrasound exposure will destroy the cell if the intensity and exposure time are beyond the tolerance limit of the culture and can even induce cell lysis leading to death [19,20] . Our data also indicate that the SonoVue microbubbles had a minimal effect on the viability of HUVECs ( Figure 2B) as it was measured at more than 90% even at high concentrations (40%, v/v).…”

Section: Resultsmentioning

confidence: 99%

Vascular gene transfer and drug delivery in vitro using low-frequency ultrasound and microbubbles

Yang

Liu

et al. 2010

Acta Pharmacol Sin

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Aim: To determine the effects of ultrasound exposure in combination with a microbubble contrast agent (SonoVue) on the cellular uptake and delivery of drugs/genes into human umbilical vein endothelial cells (HUVECs) as well as their biological effects on migration. Methods: HUVECs in suspension were exposed to pulsed ultrasound with a 10% duty cycle in combination with various concentrations of a microbubble contrast agent (SonoVue) using a digital sonifier at a frequency of 20 kHz and an intensity of 3.77 W/cm 2 on the surface of a horn tip. Cell culture inserts were used to determine the cell migration ability. Results: Exposure to pulsed ultrasound resulted in enhanced green fluorescent protein (EGFP) gene transfection efficiencies ranging from 0.2% to 2%. The transfection efficiency of HUVECs was approximately 3-fold higher in the presence of SonoVue than in its absence at the effective exposure time of 6 s. For drug delivery to HUVECs using ultrasound, the delivery efficiencies of a lowmolecular-weight model drug (TO-PRO ® -1, M W 645.38) were significantly higher when compared to drug delivery without ultrasound, with a maximum efficiency of approximately 34%. However, the delivery efficiencies of a high-molecular-weight model drug (DextranRhodamine B, M W 70 000) were low, with a maximum delivery efficiency of nearly 0.5%, and gene transfection results were similarly poor. The migration ability of HUVECs exposed to ultrasound was also lower than that of the control (no exposure). Conclusion: The use of low-frequency and low-energy ultrasound in combination with microbubbles could be a potent physical method of increasing drug/gene delivery efficiency. This technique is a promising nonviral approach that can be used in cardiovascular disease therapy.

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“…3 However, it was not focused on the challenges of increasing DNA availability from spores in environmental samples. Marentis et al 7 described a microfluidic minisonicator consisting of a channel microfabricated in glass and bonded to a quartz plate with a microfabricated gold/ZnO/gold transducer. Driving the transducer at 367 MHz transmitted acoustic waves through the quartz into the channel.…”

Section: Discussionmentioning

confidence: 99%

“…niger spore samples, used as received, were pushed through the channel and the treated samples were assessed with real-time PCR. 7 The analysis of the real-time PCR data was unconventional and the increase in DNA availability did not approach 90%.…”

Section: Discussionmentioning

confidence: 99%

A Flow-Through Ultrasonic Lysis Module for the Disruption of Bacterial Spores

Warner

Bruckner-Lea

Grate

et al. 2009

JALA: Journal of the Association for Laboratory Automation

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An automated, flow-through ultrasonic lysis module that is capable of disrupting bacterial spores to increase the DNA available for biodetection is described. The system uses a flow-through chamber that allows for direct injection of the sample without the need for a chemical or enzymatic pretreatment step to disrupt the spore coat before lysis. Lysis of Bacillus subtilis spores, a benign simulant of Bacillus anthracis, is achieved by flowing the sample through a tube whose axis is parallel to the faces of two transducers that deliver 10 W cm−2 to the surface of the tube at 1.4-MHz frequency. Increases in amplifiable DNA were assessed by real-time PCR analysis that showed at least a 25-fold increase in amplifiable DNA after ultrasonic treatment with glass beads, compared with controls with no ultrasonic power applied. The ultrasonic system and integrated fluidics are designed as a module that could be incorporated into multistep, automated sample treatment and detection systems for pathogens.

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Microfluidic sonicator for real-time disruption of eukaryotic cells and bacterial spores for DNA analysis

Cited by 62 publications

References 36 publications

The improvement of liposome-mediated transfection of pEGFP DNA into human prostate cancer cells by combining low-frequency and low-energy ultrasound with microbubbles

The improvement of liposome-mediated transfection of pEGFP DNA into human prostate cancer cells by combining low-frequency and low-energy ultrasound with microbubbles

Vascular gene transfer and drug delivery in vitro using low-frequency ultrasound and microbubbles

A Flow-Through Ultrasonic Lysis Module for the Disruption of Bacterial Spores

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