title: Real-time quantification of protein expression and translocation at individual cell resolution using imaging-dish-based live cell array.
abstract: Cell populations represent intrinsically heterogeneous systems with a high level of spatiotemporal complexity. Monitoring and understanding cell-to-cell diversity is essential for the research and application of intra- and interpopulation variations. Optical analysis of live cells is challenging since both adherent and nonadherent cells change their spatial location. However, most currently available single-cell techniques do not facilitate treatment and monitoring of the same live cells over time throughout multistep experiments. An imaging-dish-based live cell array (ID-LCA) has been developed and produced for cell handling, culturing, and imaging of numerous live cells. The dish is composed of an array of pico scale cavities-pico wells (PWs) embossed on its glass bottom. Cells are seeded, cultured, treated, and spatiotemporally measured on the ID-LCA, while each cell or small group of cells are locally constrained in the PWs. Finally, predefined cells can be retrieved for further evaluation. Various types of ID-LCAs were used in this proof-of-principle work, to demonstrate on-ID-LCA transfection of fluorescently tagged chimeric proteins, as well as the detection and kinetic analysis of their induced translocation. High variability was evident within cell populations with regard to protein expression levels as well as the extent and dynamics of protein redistribution. The association of these parameters with cell morphology and functional parameters was examined. Both the new methodology and the device facilitate research of the translocation process at individual cell resolution within large populations and thus, can potentially be used in high-throughput fashion.
authors.affiliation: The Biophysical Interdisciplinary Schottenstein Center for the Research and Technology of the Cellome, Department of Physics Building 214, Bar-Ilan University, Ramat Gan, 5290002, Israel.
authors.forename: Orit
authors.initials: O
authors.lastname: Ravid-Hermesh
authors.forename: Naomi
authors.initials: N
authors.lastname: Zurgil
authors.forename: Yana
authors.initials: Y
authors.lastname: Shafran
authors.forename: Maria
authors.initials: M
authors.lastname: Sobolev
authors.forename: Moti
authors.initials: M
authors.lastname: Galmidi
authors.forename: Yoav
authors.initials: Y
authors.lastname: Badihi
authors.forename: Liron Limor
authors.initials: LL
authors.lastname: Israel
authors.forename: Jean Paul
authors.initials: JP
authors.lastname: Lellouche
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Lellouche
authors.forename: Shulamit
authors.initials: S
authors.lastname: Michaeli
authors.forename: Mordechai
authors.initials: M
authors.lastname: Deutsch
mesh_terms: D018929:Cell Culture Techniques / Q000295:instrumentation*; D002468:Cell Physiological Phenomena; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D057054:Molecular Imaging / Q000295:instrumentation*; D011506:Proteins / Q000378:metabolism*; D059010:Single-Cell Analysis / Q000379:methods*; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism*
chemical_list: D011506:Proteins