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

Abstract

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.

Fabrication of TiO2 microspikes for highly efficient intracellular delivery by pulse laser-assisted photoporation

“…The temperature rise is signicantly high enough (>550 K) to induce cavitation bubbles at the Ti 3 O 5 /water interface. 50,74 These bubbles were generated, expanded, and destroyed, eventually resulting in strong jet ow in the vicinity of the cell membrane. As a result, the cell membranes deformed, generating transient pores which allow extracellular molecules to diffuse into the cell cytosol.…”

“…[40][41][42][43][44][45][46][47][48][49] Physical approaches are widely being developed, which use physical energy to deform the cell plasma membrane and deliver cargo into cells. 27,31,35,50 For example, in electroporation, an electric eld is required to deform cell membranes and create transient membrane pores for cargo delivery. However, it has limitations, such as electric eld distortion, pH variation, sample contamination, and high toxicity effect, reducing cell viability.…”

Microfluidic mechanoporation for cellular delivery and analysis

“…The methods are advantageous because of minimal chances of inducing harsh cellular responses such as unwanted immunogenic or chemical reactions, thereby improving delivery efficiencies. Electroporation [ [13] , [14] , [15] , [16] , [17] , [18] , [19] , [20] , [21] ], optoporation [ [22] , [23] , [24] , [25] , [26] , [27] , [28] ], magnetoporation [ [29] , [30] , [31] ], acoustoporation [ [32] , [33] , [34] , [35] ] and mechanoporation [ [36] , [37] , [38] , [39] , [40] , [41] , [42] , [43] , [44] , [45] , [46] , [47] , [48] , [49] , [50] , [51] , [52] , [53] , [54] , [55] , [56] , [57] , [58] , [59] , [60] ] are some of the most popular membrane disruption techniques. Although each of these methods have their own distinct features (elucidated in detail in Table 2 ), all of these, except mechanoporation, require an external energy field for membrane permeabilization.…”

“… Requires external energy field. >100,000 95 : 90 [ [22] , [23] , [24] , [25] , [26] , [27] , [28] ] 3. Magnetoporation - Magnetic field parameters - Vectors to permeabilize the cell membrane It can be used in vivo.…”

Single-Cell Analysis

“…The recent development of micro/nanofluidic devices can easily detect and analyze different single-cell omics such as genomics, proteomics, metabolomics as well as analyzing the different levels of disease progression [5]. Again, the devices have the ability to do single-cell therapy, diagnostics [13] and their analysis by using different physical approaches such as electroporation [14][15][16][17][18], mechanoporation [19], photoporation [20][21][22][23], microinjection [24,25] etc. Besides the extensive applications for cell separation, isolation, manipulation, and lysis, these devices are potentially helpful for studying cellular biochemical, mechanical and electrical characterization at single-cell or in subcellular level [26].…”