麻豆淫院

Many applications鈥攆rom drug discovery and diagnostics to cell engineering and gene modulation鈥攔equire delivering biomolecules into large numbers of cells and rapidly evaluating the outcomes. The challenge is two-fold: achieve intracellular delivery at scale across diverse cells and cargoes, and obtain quantitative results fast enough to keep pace with that delivery.

Researchers at the Indian Institute of Technology (IIT) Madras and Toyohashi University of Technology (TUT) have developed an integrated platform that advances both fronts simultaneously. The work is in the journal Advanced Healthcare Materials.

The platform combines two modules: a massively parallel through鈥慼ole cell鈥憇queezing mechanoporation device for high鈥憈hroughput intracellular delivery, and an automated single鈥慶ell image鈥慶ytometry pipeline built on Mask R鈥慍NN.

The device guides cells through an array of up to 62,000 tiny through鈥慼oles that are narrower than the cell diameter. A brief, gentle squeeze creates transient membrane pores that admit biomolecules into the cell interior, then reseal, allowing cells to recover.

In validation studies, the team delivered gene鈥憇ilencing RNA (siRNA) and plasmid DNA across multiple cell types, including human gingival fibroblasts (hGFs), demonstrating broad utility for cell engineering and personalized therapies.

To keep up with the fast delivery, an automated analysis system uses the same microscope images labs already acquire. A deep鈥憀earning model looks at those images and, in a single pass, reports four readouts: cell size, the fraction of cargo鈥憄ositive cells, the fraction of viable cells, and per鈥慶ell fluorescence intensity.

What used to take hours of manual counting now takes about 83 seconds on a representative dataset of roughly 500 cells, with accuracy comparable to human review.

Large cohorts were processed鈥攆or example, 1,980 cells (6鈥慒AM siRNA) and 1,184 (EGFP plasmid). These sample sizes make the results statistically robust and turn high鈥憈hroughput delivery into high鈥慶onfidence decisions.

"Our goal was simple: get molecules inside many cells quickly and gently," says first author Pulasta Chakrabarty. "Seeing the device work across different cell types points to real potential in cell engineering and personalized therapies."

Co鈥慳uthor Ryoma Suzuki adds, "The automated model looks at the same images labs already use. It does the counting and measuring鈥攃ell size, delivery efficiency, viability, and per鈥慶ell fluorescence鈥攊n one pass, so the evaluation keeps pace with the experiments."

"High throughput alone isn't enough," notes Prof. Moeto Nagai. "What matters is the speed at which results can be trusted. By unifying high鈥憈hroughput delivery with automated quality control, we move from proofs鈥憃f鈥慶oncept to practical workflows鈥攁nd closer to systems that prepare a patient's cells on鈥憇ite."

By coupling scalable intracellular delivery with rapid, automated evaluation, the platform expands what can be accomplished in a single day鈥攆rom large鈥憇cale screening and diagnostics to practical cell manipulation and future point鈥憃f鈥慶are gene鈥慹diting workflows.