Version 2 2022-06-03, 17:04Version 2 2022-06-03, 17:04
Version 1 2022-05-23, 16:11Version 1 2022-05-23, 16:11
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posted on 2022-06-03, 17:04authored byCyril Deroy, James H. R. Wheeler, Agata N. Rumianek, Peter R. Cook, William M. Durham, Kevin R. Foster, Edmond J. Walsh
Microfluidic devices
are widely used in many fields of biology,
but a key limitation is that cells are typically surrounded by solid
walls, making it hard to access those that exhibit a specific phenotype
for further study. Here, we provide a general and flexible solution
to this problem that exploits the remarkable properties of microfluidic
circuits with fluid wallstransparent interfaces between culture
media and an immiscible fluorocarbon that are easily pierced with
pipets. We provide two proofs of concept in which specific cell subpopulations
are isolated and recovered: (i) murine macrophages chemotaxing toward
complement component 5a and (ii) bacteria (Pseudomonas
aeruginosa) in developing biofilms that migrate toward
antibiotics. We build circuits in minutes on standard Petri dishes,
add cells, pump in laminar streams so molecular diffusion creates
attractant gradients, acquire time-lapse images, and isolate desired
subpopulations in real time by building fluid walls around migrating
cells with an accuracy of tens of micrometers using 3D printed adaptors
that convert conventional microscopes into wall-building machines.
Our method allows live cells of interest to be easily extracted from
microfluidic devices for downstream analyses.