Nano-localized single-cell nano-electroporation

Lab on a Chip ◽  
2020 ◽  
Vol 20 (22) ◽  
pp. 4194-4204
Author(s):  
Tuhin Subhra Santra ◽  
Srabani Kar ◽  
Hwan-You Chang ◽  
Fan-Gang Tseng
Keyword(s):  
Cell Viability ◽  
Single Cell ◽  
Intracellular Delivery ◽  
High Cell ◽  
Highly Efficient

We demonstrated nano-electroporation technique to create transient nano-holes at single or multiple nano-localized positions of a single-cell for a highly efficient intracellular delivery with high cell viability.

Near-infrared nanosecond-pulsed laser-activated highly efficient intracellular delivery mediated by nano-corrugated mushroom-shaped gold-coated polystyrene nanoparticles

Nanoscale ◽  
2020 ◽  
Vol 12 (22) ◽  
pp. 12057-12067 ◽  
Author(s):  
Tuhin Subhra Santra ◽  
Srabani Kar ◽  
Te-Chang Chen ◽  
Chih-Wei Chen ◽  
Jayant Borana ◽  
...  
Keyword(s):  
Cell Viability ◽  
Near Infrared ◽  
Pulsed Laser ◽  
Intracellular Delivery ◽  
High Cell ◽  
Polystyrene Nanoparticles ◽  
Infrared Wavelength ◽  
Highly Efficient ◽  
Nanosecond Pulsed Laser

Here, an efficient intracellular delivery of molecules with high cell viability is reported using plasmonic photoporation mediated by nano-corrugated mushroom-shaped gold-coated polystyrene nanoparticles at near-infrared wavelength.

scholarly journals Efficient Intracellular Delivery of Molecules with High Cell Viability Using Nanosecond-Pulsed Laser-Activated Carbon Nanoparticles

ACS Nano ◽  
2014 ◽  
Vol 8 (3) ◽  
pp. 2889-2899 ◽  
Author(s):  
Aritra Sengupta ◽  
Sean C. Kelly ◽  
Nishant Dwivedi ◽  
Naresh Thadhani ◽  
Mark R. Prausnitz
Keyword(s):  
Activated Carbon ◽  
Cell Viability ◽  
Carbon Nanoparticles ◽  
Pulsed Laser ◽  
Intracellular Delivery ◽  
High Cell ◽  
Nanosecond Pulsed Laser

Nanolocalized Single-Cell-Membrane Nanoelectroporation: For higher efficiency with high cell viability.

2014 ◽  
Vol 8 (1) ◽  
pp. 30-34 ◽  
Author(s):  
Tuhin Subhra Santra ◽  
Srabani Kar ◽  
Jayant Borana ◽  
Pen-Cheng Wang ◽  
Fan-Gang Tseng
Keyword(s):  
Cell Membrane ◽  
Cell Viability ◽  
Single Cell ◽  
High Cell

scholarly journals Systematic Study of Single-Cell Isolation from Musculoskeletal Tissues for Single-Cell Sequencing

2020 ◽  
Author(s):  
Peng Guo ◽  
Xizhe Liu ◽  
Penghui Zhang ◽  
Zhongyuan He ◽  
Liru Wen ◽  
...  
Keyword(s):  
Quality Control ◽  
Articular Cartilage ◽  
Cell Viability ◽  
Single Cell ◽  
Nucleus Pulposus ◽  
Posterior Longitudinal Ligament ◽  
Cell Aggregates ◽  
High Cell ◽  
Musculoskeletal Tissues ◽  
Single Cell Isolation

Abstract Background: The single-cell platform provided revolutionary way to study cellular biology. Technologically, a sophistic protocol of isolating qualified single cells would be key to deliver to single-cell platform, which requires high cell viability, high cell yield and low content of cell aggregates or doublets. For musculoskeletal tissues, like bone, cartilage, nucleus pulposus, tendons, etc. as well as their pathological state, which are tense and dense, it’s full of challenge to efficiently and rapidly prepare qualified single-cell suspension. Conventionally, enzymatic dissociation methods were wildly used but lack of quality control. In the present study, we designed specific enzymatic treatment protocols for several human pathological musculoskeletal tissues, including degenerated nucleus pulposus, ossifying posterior longitudinal ligament and knee articular cartilage with osteoarthritis, aiming to rapidly and efficiently harvest qualified single-cell suspensions to meet the requirements of single-cell RNA-sequencing (scRNA-seq).Results: The single-cell suspensions from human degenerated nucleus pulposus and ossifying posterior longitudinal ligaments were both qualified after systematic quality control. Bioanalyzer trace showed expected cDNA size distribution of the scRNA-seq library. A clear separation of cellular barcodes from background partitions were verified by the barcode-rank plot after sequencing. However, we failed to obtain eligible samples from articular cartilage due to low cell viability and excessive cell aggregates and doublets. Conclusions: In conclusion, we provided rapid and efficient single-cell isolation protocols for human degenerated nucleus pulposus and ossifying posterior longitudinal ligament, which could be applied for scRNA-seq. More efforts will be made on improving the protocols for human articular cartilage.

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

RSC Advances ◽  
2021 ◽  
Vol 11 (16) ◽  
pp. 9336-9348
Author(s):  
L. Mohan ◽  
Srabani Kar ◽  
Pallab Sinha Mahapatra ◽  
Moeto Nagai ◽  
Tuhin Subhra Santra
Keyword(s):  
Cell Viability ◽  
Pulse Laser ◽  
Intracellular Delivery ◽  
Living Cells ◽  
Highly Efficient ◽  
Delivery Efficiency

The introduction of foreign cargo into living cells with high delivery efficiency and cell viability by laser asisted photoporation on TiO2 microspikes platform.

Probing Cellular Outcomes Using Heterobivalent Constructs

2019 ◽  
Author(s):  
Rohit Bhadoria ◽  
Kefeng Ping ◽  
Christer Lohk ◽  
Ivar Järving ◽  
Pavel Starkov
Keyword(s):  
Cell Viability ◽  
Small Molecules ◽  
Cellular Uptake ◽  
Kinase Inhibitors ◽  
Rho Kinase ◽  
Intracellular Delivery ◽  
Rho Kinase Inhibitors

<div> <div> <div> <p>Conjugation techniques are central to improving intracellular delivery of bioactive small molecules. However, tracking and assessing the overall biological outcome of these constructs remains poorly understood. We addressed this issue by having developed a focused library of heterobivalent constructs based on Rho kinase inhibitors to probe various scenarios. By comparing induction of a phenotype of interest vs. cell viability vs. cellular uptake, we demonstrate that such conjugates indeed lead to divergent cellular outcomes. </p> </div> </div> </div>

scholarly journals Microfluidic Based Physical Approaches towards Single-Cell Intracellular Delivery and Analysis

Micromachines ◽  
2021 ◽  
Vol 12 (6) ◽  
pp. 631
Author(s):  
Kiran Kaladharan ◽  
Ashish Kumar ◽  
Pallavi Gupta ◽  
Kavitha Illath ◽  
Tuhin Subhra Santra ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Biology ◽  
Transfection Efficiency ◽  
Intracellular Delivery ◽  
Diagnostic Tools ◽  
Delivery Methods ◽  
Physical Methods ◽  
Cell Measurement ◽  
Therapeutic Development ◽  
Single Living

The ability to deliver foreign molecules into a single living cell with high transfection efficiency and high cell viability is of great interest in cell biology for applications in therapeutic development, diagnostics, and drug delivery towards personalized medicine. Various physical delivery methods have long demonstrated the ability to deliver cargo molecules directly to the cytoplasm or nucleus and the mechanisms underlying most of the approaches have been extensively investigated. However, most of these techniques are bulk approaches that are cell-specific and have low throughput delivery. In comparison to bulk measurements, single-cell measurement technologies can provide a better understanding of the interactions among molecules, organelles, cells, and the microenvironment, which can aid in the development of therapeutics and diagnostic tools. To elucidate distinct responses during cell genetic modification, methods to achieve transfection at the single-cell level are of great interest. In recent years, single-cell technologies have become increasingly robust and accessible, although limitations exist. This review article aims to cover various microfluidic-based physical methods for single-cell intracellular delivery such as electroporation, mechanoporation, microinjection, sonoporation, optoporation, magnetoporation, and thermoporation and their analysis. The mechanisms of various physical methods, their applications, limitations, and prospects are also elaborated.

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