Micromolded arrays for separation of adherent cells.
We present an efficient, yet inexpensive, approach for isolating viable single cells or colonies from a mixed population. This cell microarray platform possesses innovations in both the array manufacture and the manner of target cell release. Arrays of microwells with bases composed of detachable concave elements, termed microrafts, were fabricated by a dip-coating process using a polydimethylsiloxane mold as the template and the array substrate. This manufacturing approach enabled the use of materials other than photoresists to create the array elements. Thus microrafts possessing low autofluorescence could be fabricated for fluorescence-based identification of cells. Cells plated on the microarray settled and attached at the center of the wells due to the microrafts' concavity. Individual microrafts were readily dislodged by the action of a needle inserted through the compliant polymer substrate. The hard polymer material (polystyrene or epoxy resin) of which the microrafts were composed protected the cells from damage by the needle. For cell analysis and isolation, cells of interest were identified using a standard inverted microscope and microrafts carrying target cells were dislodged with the needle. The released cells/microrafts could be efficiently collected, cultured and clonally expanded. During the separation and collection procedures, the cells remained adherent and provided a measure of protection during manipulation, thus providing an extremely high single-cell cloning rate (>95%). Generation of a transfected cell line based on expression of a fluorescent protein demonstrated an important application for performing on-chip cell separations.
Wang
Yuli
Y
Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599, USA.
Phillips
Colleen
C
Xu
Wei
W
Pai
Jeng-Hao
JH
Dhopeshwarkar
Rahul
R
Sims
Christopher E
CE
Allbritton
Nancy
N
D002448:Cell Adhesion; D002469:Cell Separation / Q000295:instrumentation*; D004129:Dimethylpolysiloxanes; D005109:Extracellular Matrix; D005453:Fluorescence; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans
D004129:Dimethylpolysiloxanes; D049452:Green Fluorescent Proteins; C013830:baysilon