Infrared Pulse Laser-Activated Highly Efficient Intracellular Delivery Using Titanium Microdish Device.

We report infrared (IR) pulse laser-activated highly efficient parallel intracellular delivery by using an array of titanium microdish (TMD) device. Upon IR laser pulse irradiation, a two-dimensional array of TMD device generated photothermal cavitation bubbles to disrupt the cell membrane surface and create transient membrane pores to deliver biomolecules into cells by a simple diffusion process. We successfully delivered the dyes and different sizes of dextran in different cell types with variations of laser pulses. Our platform has the ability to transfect more than a million cells in a parallel fashion within a minute. The best results were achieved for SiHa cells with a delivery efficiency of 96% and a cell viability of around 98% for propidium iodide dye using 600 pulses, whereas a delivery efficiency of 98% and a cell viability of 100% were obtained for dextran 3000 MW delivery using 700 pulses. For dextran 10,000 MW, the delivery efficiency was 92% and the cell viability was 98%, respectively. The device is compact, easy-to-use, and potentially applicable for cellular therapy and diagnostic purposes.

[1]  T. Flaig,et al.  Multifunctional nanoclusters of NaYF4:Yb3+,Er3+ upconversion nanoparticle and gold nanorod for simultaneous imaging and targeted chemotherapy of bladder cancer. , 2019, Materials science & engineering. C, Materials for biological applications.

[2]  Xitao Wang,et al.  Enhanced thermal conductivity in copper matrix composites reinforced with titanium-coated diamond particles , 2011 .

[3]  Z. Hartman,et al.  Adenovirus vector induced innate immune responses: impact upon efficacy and toxicity in gene therapy and vaccine applications. , 2008, Virus research.

[4]  Fan-Gang Tseng,et al.  Recent Trends on Micro/Nanofluidic Single Cell Electroporation , 2013, Micromachines.

[5]  N. Melosh,et al.  Universal intracellular biomolecule delivery with precise dosage control , 2018, Science Advances.

[6]  Moon J. Kim,et al.  Effect of Ti interlayer on interfacial thermal conductance between Cu and diamond , 2018, Acta Materialia.

[7]  Xinqi Fan,et al.  Near-Infrared Light Activation of Proteins Inside Living Cells Enabled by Carbon Nanotube-Mediated Intracellular Delivery. , 2016, ACS applied materials & interfaces.

[8]  Alexander A. Oraevsky,et al.  Laser activated nanothermolysis of leukemia cells monitored by photothermal microscopy , 2005, SPIE BiOS.

[9]  Ronald G. Crystal,et al.  Transfer of Genes to Humans: Early Lessons and Obstacles to Success , 1995, Science.

[10]  W. D. De Vos,et al.  Fast spatial‐selective delivery into live cells , 2017, Journal of controlled release : official journal of the Controlled Release Society.

[11]  M. C. Mancini,et al.  Bioimaging: second window for in vivo imaging. , 2009, Nature nanotechnology.

[12]  L. Bonacina,et al.  Live cells assessment of opto-poration by a single femtosecond temporal Airy laser pulse , 2018, AIP Advances.

[13]  Nicole Rusk Torrents of sequence , 2011, Nature Methods.

[14]  Ting-Hsiang Wu,et al.  Massively parallel delivery of large cargo into mammalian cells with light pulses , 2015, Nature Methods.

[15]  Giovanna Zinzalla,et al.  Targeting protein-protein interactions for therapeutic intervention: a challenge for the future. , 2009, Future medicinal chemistry.

[16]  Nam-Trung Nguyen,et al.  Design, fabrication and characterization of drug delivery systems based on lab-on-a-chip technology. , 2013, Advanced drug delivery reviews.

[17]  F. Tantussi,et al.  Soft electroporation for delivering molecules into tightly adherent mammalian cells through 3D hollow nanoelectrodes , 2017, Scientific Reports.

[18]  Lior Pachter,et al.  Single-cell analysis at the threshold , 2016, Nature Biotechnology.

[19]  C T W Moonen,et al.  Understanding ultrasound induced sonoporation: definitions and underlying mechanisms. , 2014, Advanced drug delivery reviews.

[20]  Fan-Gang Tseng,et al.  Tuning nano electric field to affect restrictive membrane area on localized single cell nano-electroporation , 2013 .

[21]  X. Ruan,et al.  First principles calculation of lattice thermal conductivity of metals considering phonon-phonon and phonon-electron scattering , 2016 .

[22]  R. Herzog,et al.  Progress and prospects: immune responses to viral vectors , 2010, Gene Therapy.

[23]  D. Miklavčič,et al.  Electroporation in dense cell suspension--theoretical and experimental analysis of ion diffusion and cell permeabilization. , 2007, Biochimica et biophysica acta.

[24]  Thierry Biben,et al.  Threshold for Vapor Nanobubble Generation Around Plasmonic Nanoparticles , 2017 .

[25]  Fan-Gang Tseng,et al.  Essentials of Single-Cell Analysis , 2016 .

[26]  Pallavi Shinde,et al.  Current Trends of Microfluidic Single-Cell Technologies , 2018, International journal of molecular sciences.

[27]  D. Werner,et al.  Improved Working Model for Interpreting the Excitation Wavelength- and Fluence-Dependent Response in Pulsed Laser-Induced Size Reduction of Aqueous Gold Nanoparticles , 2011 .

[28]  Alexander A. Oraevsky,et al.  Elimination of leukemic cells from human transplants by laser nano-thermolysis , 2006, SPIE BiOS.

[29]  R. Weissleder A clearer vision for in vivo imaging , 2001, Nature Biotechnology.

[30]  M. Meunier,et al.  Photothermal response of hollow gold nanoshell to laser irradiation: Continuous wave, short and ultrashort pulse , 2015 .

[31]  R. Schmid,et al.  Liposome mediated gene transfer into the rat oesophagus , 1997, Gut.

[32]  Pallavi Shinde,et al.  Single-cell electroporation: current trends, applications and future prospects , 2018, Journal of Micromechanics and Microengineering.

[33]  Eric Mazur,et al.  Intracellular Delivery Using Nanosecond-Laser Excitation of Large-Area Plasmonic Substrates. , 2017, ACS nano.