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Biology subjects

Castrejon-Pita, A. A.

Publications and source records attributed to Castrejon-Pita, A. A..

2 recordsLinked to original sources

Identification and isolation of migratory subpopulations of cancer cells using microfluidics with fluid walls

Microfluidic assays have great potential for investigating cancer cell migration, a key process underlying metastasis which remains a major clinical challenge. However, adoption by biologists is limited due to complex fabrication and operation methods combined with difficulty in retrieving cells of interest. We jet-print dumbbell-shaped microfluidic environments with fluid walls in standard Petri dishes in seconds using standard growth media and immiscible oil. Prostate cancer cells and chemoattractant (bovine serum) are introduced into each end using local transient differences in Laplace pressure to enable spatial confinement. Automated live-cell imaging monitors chemotaxis towards serum using standard microscopes, as the confining fluid walls are optically transparent. Distinct subpopulations of migratory cells exhibit different morphologies and motilities, assessed using a custom machine-learning segmentation model. Then, we leverage the reconfigurability of fluid walls to isolate and retrieve rapidly-migrating single cells. This demonstrates that fluid-walled microfluidics is a versatile and accessible approach for investigating cancer cell migration at single-cell resolution, offering new opportunities to examine cellular heterogeneity in metastasis.

bioengineering↗

Creating wounds in cell monolayers using micro-jets

Many wound-healing assays are used in cell biology and biomedicine; they are often labor intensive and/or require specialized and costly equipment. We describe a contactless method to create wounds with any imaginable 2D pattern in cell monolayers using micro-jets of either media or an immiscible and biocompatible fluorocarbon (i.e., FC40). We also combine this with another method that allows automation and multiplexing using standard Petri dishes. A dish is filled with a thin film of media overlaid with FC40, and the two liquids reshaped into an array of microchambers in minutes. Each chamber in such a grid is isolated from others by fluid walls of FC40. Cells are now added, allowed to grow into a monolayer, and wounds created using the microjets; then, healing is monitored by microscopy. As arrays of chambers can be made using the media and Petri dishes familiar to biologists, and as dishes fit seamlessly into their incubators, microscopes, and workflows, we anticipate this assay will find wide application in biomedicine.

bioengineering↗