Abstract.
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-µm-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.
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Wakamoto, Y., Inoue, I., Moriguchi, H. et al. Analysis of single-cell differences by use of an on-chip microculture system and optical trapping. Fresenius J Anal Chem 371, 276–281 (2001). https://doi.org/10.1007/s002160100999
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DOI: https://doi.org/10.1007/s002160100999