Selective chemical treatment of cellular microdomains using multiple laminar streams.
There are many experiments in which it would be useful to treat a part of the surface or interior of a cell with a biochemical reagent. It is difficult, however, to achieve subcellular specificity, because small molecules diffuse distances equal to the extent of the cell in seconds. This paper demonstrates experimentally, and analyzes theoretically, the use of multiple laminar fluid streams in microfluidic channels to deliver reagents to, and remove them from, cells with subcellular spatial selectivity. The technique made it possible to label different subpopulations of mitochondria fluorescently, to disrupt selected regions of the cytoskeleton chemically, to dislodge limited areas of cell-substrate adhesions enzymatically, and to observe microcompartmental endocytosis within individual cells. This technique does not require microinjection or immobilization of reagents onto nondiffusive objects; it opens a new window into cell biology.
Takayama
Shuichi
S
Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, MA 02138, USA.
Ostuni
Emanuele
E
LeDuc
Philip
P
Naruse
Keiji
K
Ingber
Donald E
DE
Whitesides
George M
GM
D000818:Animals; D001426:Bacterial Proteins; D002417:Cattle; D002478:Cells, Cultured; D003584:Cytological Techniques; D003599:Cytoskeleton / Q000737:chemistry; D004705:Endocytosis / Q000187:drug effects; D005456:Fluorescent Dyes; D007091:Image Processing, Computer-Assisted; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D008928:Mitochondria / Q000737:chemistry; D008954:Models, Biological; D036103:Nanotechnology; D010539:Permeability; D011956:Receptors, Cell Surface / Q000502:physiology; D013347:Subcellular Fractions / Q000737:chemistry / Q000187:drug effects*; D014162:Transfection; D014357:Trypsin / Q000737:chemistry
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D011956:Receptors, Cell Surface; C054562:yellow fluorescent protein, Bacteria; D014357:Trypsin