Figure 1: End‑to‑end workflow: 2D cell squeezing for intracellular delivery and AI image cytometry for single‑cell readout (IMAGE)
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Top row (delivery and imaging). Cells and cargo are introduced into a microfluidic chip whose base integrates a porous SU‑8 membrane with vertical through‑holes and a PDMS chamber. As individual cells pass the sub‑cell‑sized through-holes, a brief 2D squeeze forms transient membrane pores, allowing cargo to enter; after passage, the membrane reseals, and cells exit to the outlets. Treated cells are then collected and imaged under a microscope (bright‑field and fluorescence).
Bottom row (automated single‑cell analysis). The same image set is processed by image cytometry using a Mask R‑CNN–based instance‑segmentation model: (i) bright‑field for cell localization, (ii) green fluorescence for cargo delivery, (iii) red fluorescence for viability/dying cells (e.g., PI or Calcein red‑orange), and (iv) instance‑segmentation overlays for per‑cell measurements. A simple rule-based map classifies each cell by the presence/absence of green and red signals into four states (delivered‑live, delivered‑dead, undelivered‑live, undelivered‑dead), enabling automatic calculation of delivery efficiency, viability, and per‑cell delivered amount. The diameter of each cell in an image is evaluated from the instance segmentation masks.
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