Analysis of single-cell differences by use of an on-chip microculture system and optical trapping.
A method is described for continuous observation of isolated single cells that enables genetically identical cells to be compared; it uses an on-chip microculture system and optical tweezers. Photolithography is used to construct microchambers with 5-microm-high walls made of thick photoresist (SU-8) on the surface of a glass slide. These microchambers are connected by a channel through which cells are transported, by means of optical tweezers, from a cultivation microchamber to an analysis microchamber, or from the analysis microchamber to a waste microchamber. The microchambers are covered with a semi-permeable membrane to separate them from nutrient medium circulating through a "cover chamber" above. Differential analysis of isolated direct descendants of single cells showed that this system could be used to compare genetically identical cells under contamination-free conditions. It should thus help in the clarification of heterogeneous phenomena, for example unequal cell division and cell differentiation.
Wakamoto
Y
Y
Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Japan.
Inoue
I
I
Moriguchi
H
H
Yasuda
K
K
D001419:Bacteria / Q000254:growth & development*; D019149:Bioreactors; D002455:Cell Division; D002999:Clone Cells / Q000166:cytology; D015194:Diffusion Chambers, Culture / Q000379:methods*; D004852:Epoxy Compounds; D004926:Escherichia coli / Q000254:growth & development; D007834:Lasers; D008846:Micromanipulation / Q000379:methods*; D011108:Polymers
D004852:Epoxy Compounds; D011108:Polymers; C520895:SU-8 compound