Propidium iodide labeling of nanoparticles as a novel tool for the quantification of cellular binding and uptake

Neumeyer, Andrea; Bukowski, Mirko; Veith, Michael; Lehr, Claus‐Michael; Daum, Nicole

doi:10.1016/j.nano.2010.12.007

Cited by 27 publications

(25 citation statements)

References 48 publications

(48 reference statements)

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“…However these methods are either tedious or cannot be used with all types of particles. 49 In this work, flow cytometry to was used demonstrate the efficiency of the magnetofection with SPIONS.…”

Section: Discussionmentioning

confidence: 99%

Electrostatic assembly of a DNA superparamagnetic nano-tool for simultaneous intracellular delivery and in situ monitoring

Geinguenaud

Souissi

Fagard

et al. 2012

Nanomedicine: Nanotechnology, Biology and Medicine

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Section: Discussionmentioning

confidence: 99%

Electrostatic assembly of a DNA superparamagnetic nano-tool for simultaneous intracellular delivery and in situ monitoring

Geinguenaud

Souissi

Fagard

et al. 2012

Nanomedicine: Nanotechnology, Biology and Medicine

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“…32 CSAgNPs dispersed in DMEM were mixed with PI (0.1 µg/mL) and the mixture was incubated at room temperature for 2 hours. RAW264.7 cells (1 × 10 5 cells/mL) were grown on glass coverslips.…”

Section: Cellular Uptake Studiesmentioning

confidence: 99%

Toxicity and antibacterial assessment of chitosan-coated silver nanoparticles on human pathogens and macrophage cells

Jena¹,

Mohanty²,

Mallick³

et al. 2012

IJN

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Background: Pathogenic bacteria are able to develop various strategies to counteract the bactericidal action of antibiotics. Silver nanoparticles (AgNPs) have emerged as a potential alternative to conventional antibiotics because of their potent antimicrobial properties. The purpose of this study was to synthesize chitosan-stabilized AgNPs (CS-AgNPs) and test for their cytotoxic, genotoxic, macrophage cell uptake, antibacterial, and antibiofilm activities. Methods: AgNPs were synthesized using chitosan as both a stabilizing and a reducing agent. Antibacterial activity was determined by colony-forming unit assay and scanning electron microscopy. Genotoxic and cytotoxic activity were determined by DNA fragmentation, comet, and MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assays. Cellular uptake and intracellular antibacterial activity were tested on macrophages. Results: CS-AgNPs exhibited potent antibacterial activity against different human pathogens and also impeded bacterial biofilm formation. Scanning electron microscopy analysis indicated that CS-AgNPs kill bacteria by disrupting the cell membrane. CS-AgNPs showed no significant cytotoxic or DNA damage effect on macrophages at the bactericidal dose. Propidium iodide staining indicated active endocytosis of CS-AgNPs resulting in reduced intracellular bacterial survival in macrophages. Conclusion:The present study concludes that at a specific dose, chitosan-based AgNPs kill bacteria without harming the host cells, thus representing a potential template for the design of antibacterial agents to decrease bacterial colonization and to overcome the problem of drug resistance.

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“…PI (Sigma) was used in this experiment as an injection particle to determine the efficiency and viability of the lance array nanoinjection method. When PI comes into contact with genetic material, PI molecules intercalate between base pairs of DNA and ribonucleic acid (RNA) [25][26][27][28][29][30][31]. PI molecules attached to nucleic acids emit enhanced (20x-30x) fluorescent light waves when excited by certain lamps, making PI positive cells detectable with flow cytometry.…”

Section: Lance Array Fabrication and Injection Devicementioning

confidence: 99%

“…PI molecules attached to nucleic acids emit enhanced (20x-30x) fluorescent light waves when excited by certain lamps, making PI positive cells detectable with flow cytometry. PI does not penetrate the membrane of viable cells [26,30,31,32], so PI positive cells in this experiment must be either successfully injected, dead, or both. In previous experi ments, no cytotoxicity has been seen in cells for at least 1 h after the addition of PI to islet cells [33].…”

Section: Lance Array Fabrication and Injection Devicementioning

confidence: 99%

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

Lindstrom

Brewer

Ferguson

et al. 2014

Journal of Nanotechnology in Engineering and Medicine

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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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Propidium iodide labeling of nanoparticles as a novel tool for the quantification of cellular binding and uptake

Cited by 27 publications

References 48 publications

Electrostatic assembly of a DNA superparamagnetic nano-tool for simultaneous intracellular delivery and in situ monitoring

Electrostatic assembly of a DNA superparamagnetic nano-tool for simultaneous intracellular delivery and in situ monitoring

Toxicity and antibacterial assessment of chitosan-coated silver nanoparticles on human pathogens and macrophage cells

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

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