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Research Papers

Injection of Propidium Iodide into HeLa Cells Using a Silicon Nanoinjection Lance Array

[+] Author and Article Information
Zachary K. Lindstrom, Steven J. Brewer, Melanie A. Ferguson

Department of Mechanical Engineering,
Brigham Young University,
Provo, UT 84602

Sandra H. Burnett

Department of Microbiology
and Molecular Biology,
Brigham Young University,
Provo, UT 84602

Brian D. Jensen

Department of Mechanical Engineering,
Brigham Young University,
Provo, UT 84602
e-mail: bdjensen@byu.edu

1Corresponding author.

Manuscript received April 4, 2014; final manuscript received September 15, 2014; published online October 3, 2014. Assoc. Editor: Malisa Sarntinoranont.

J. Nanotechnol. Eng. Med 5(2), 021008 (Oct 03, 2014) (7 pages) Paper No: NANO-14-1028; doi: 10.1115/1.4028603 History: Received April 04, 2014; Revised September 15, 2014

Delivering foreign molecules into human cells is a wide and ongoing area of research. Gene therapy, or delivering nucleic acids into cells via nonviral or viral pathways, is an especially promising area for pharmaceutics. All gene therapy methods have their respective advantages and disadvantages, including limited delivery efficiency and low viability. We present an electromechanical method for delivering foreign molecules into human cells. Nanoinjection, or delivering molecules into cells using a solid lance, has proven to be highly efficient while maintaining high viability levels. This paper describes an array of solid silicon microlances that was tested to determine efficiency and viability when nanoinjecting tens of thousands of HeLa cells simultaneously. Propidium iodide (PI), a dye that fluoresces when bound to nucleic acids and does not fluoresce when unbound, was delivered into cells using the lance array. Results show that the lance array delivers PI into up to 78% of a nanoinjected HeLa cell culture, while maintaining 78–91% viability. With these results, we submit the nanoinjection method using a silicon lance array as another promising particle delivery method for mammalian culture cells.

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Figures

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Fig. 2

Images of the injection device from (a) above and (b) below

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Fig. 1

Scanning electron micrograph showing the edge of a lance array chip

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Fig. 3

Optical micrograph of a HeLa 229 cell culture. The cells are approximately 50% confluent.

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Fig. 4

Schematic of the setup for the nanoinjection process with a lance array, using the application of an electric field (not to scale)

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Fig. 5

Process diagram for the nanoinjection technique using a lance array and voltage (not to scale). The process begins with (a) cultured cells in solution with PI and the lance array, with a negative voltage applied to the lances to accumulate PI molecules. The lances penetrate cell membranes (b), and the voltage is reversed (c), releasing the PI molecules inside the cytoplasm and/or nucleus of cells. The lances are removed from the cells (d), leaving PI molecules inside.

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Fig. 7

Bar graph comparing average viability data for controls, diffusion injections, 1.5 VDC injections, and pulsed voltage injections. The error bars indicate a 95% confidence interval (α = 0.05). *Statistically significant difference in population averages (p < 0.05).

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Fig. 6

Bar graph comparing average PI uptake data for controls, diffusion injections, 1.5 VDC injections, and pulsed voltage injections. The error bars indicate a 95% confidence interval (α = 0.05). *Statistically significant difference in population averages (p < 0.05).

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Fig. 10

Bar graph comparing PI uptake between controls and cells nanoinjected with PI added at several time intervals. Error bars indicate a 95% confidence interval (α = 0.05). *Statistically significant difference in population averages (p < 0.05).

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Fig. 11

Bar graph comparing viability between controls and cells nanoinjected with PI added at several time intervals. Error bars indicate a 95% confidence interval (α = 0.05).

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Fig. 8

Bar graph comparing average viability data for controls and cells punctured with the lance array. The error bars indicate a 95% confidence interval (α = 0.05). *Statistically significant difference in population averages (p < 0.05).

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Fig. 9

Bar graph comparing PI uptake between cells strained with a bare silicon chip, cells punctured with lances, and unpunctured controls. Error bars indicate a 95% confidence interval (α = 0.05). **Statistically significant difference in population averages (p < 0.01).

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