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Related Concept Videos

Subcellular Fractionation01:32

Subcellular Fractionation

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The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
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Updated: May 6, 2026

Cell Squeezing as a Robust, Microfluidic Intracellular Delivery Platform
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High Throughput Intracellular Delivery Using a 2D Cell-Squeezing Mechanoporation Device and Its Analysis by a Deep

Pulasta Chakrabarty1, Abinaya R1, Ryoma Suzuki2

  • 1Department of Engineering Design, Indian Institute of Technology Madras, Chennai, Tamil Nadu, 600036, India.

Advanced Healthcare Materials
|August 21, 2025
PubMed
Summary
This summary is machine-generated.

A novel 2D microfluidic device enhances cell-squeezing mechanoporation for high-throughput intracellular delivery. This technology efficiently delivers molecules like dextran and siRNA into various cell types, enabling advanced research and therapeutic applications.

Keywords:
cell‐squeezingdeep learningmechanoporationmicrofluidicssingle‐cell

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Area of Science:

  • Biotechnology
  • Microfluidics
  • Cell Biology

Background:

  • Cell-squeezing mechanoporation offers intracellular delivery but faces throughput limitations with 1D constrictions.
  • Developing high-throughput methods is crucial for advancing single-cell delivery applications.

Purpose of the Study:

  • To design and fabricate a 2D microfluidic device for parallel, high-throughput cell-squeezing mechanoporation.
  • To demonstrate efficient intracellular delivery of various molecules into diverse cell types using the novel device.

Main Methods:

  • Fabrication of a 2D microfluidic device with an array of vertical through-holes (8-15 µm diameter) on a thin SU-8 membrane within a PDMS structure.
  • Intracellular delivery via rapid cell shearing through microfluidic constrictions, enabling diffusion-based molecule uptake.
  • Analysis of delivery efficiency using image cytometry, combining instance segmentation and rule-based image processing for automated single-cell state evaluation.

Main Results:

  • Achieved high-throughput dextran (4-40 kDa) delivery into HeLa and Jurkat cells at rates up to 3 million cells/min.
  • Successfully delivered small interfering ribonucleic acid (siRNA) and plasmids into primary human mesenchymal stem cells (hMSCs) and human gingival fibroblasts (hGFs).
  • Automated deep learning-based analysis system enabled single-cell resolution quantification of mechanoporation-based intracellular delivery.

Conclusions:

  • The 2D cell-squeezing microfluidic device significantly increases throughput for intracellular delivery compared to 1D methods.
  • The platform demonstrates broad applicability for delivering molecules and genetic material into various cell types, including primary cells.
  • The integrated automated analysis system provides precise, single-cell resolution quantification, validating the device's potential for therapeutic applications.