title: Integrated optical molecular imaging system for four-dimensional real-time detection in living single cells.
abstract: A novel, multifunctional optical imaging system was developed by integrating four-dimensional (4D) real-time confocal microscopy (RT-CM), multicolor total internal reflection microscopy (TIRFM), and Nomarski differential interference contrast (DIC) microscopy based on an epifluorescence microscope platform. A microcell incubator was combined with the imaging system for extended, real-time monitoring of living cells. The 4D images were generated by a combination of 3D images and multiple spatial or time images of a specimen, obtained at 10 nm intervals. Optical sectioning was accomplished with a z-motor, which obtained 4D information with sequential layered sections. The integrated imaging system showed excellent detection sensitivity at the single-molecule level and 3D-spatial resolution (20 nm x-y and 10 nm z-axis) without moving the cell sample. This could be a tool for obtaining crucial information needed to develop approaches for characterizing and understanding the dynamics of biomolecules and nanoparticles in individual living cells and molecular interactions at the single-molecule level.
authors.affiliation: Department of Applied Chemistry, College of Applied Science, Kyung Hee University, Yongin-si, Gyeonggi-do 446-701, South Korea.
authors.forename: Seungah
authors.initials: S
authors.lastname: Lee
authors.forename: Seong Ho
authors.initials: SH
authors.lastname: Kang
mesh_terms: D000818:Animals; D002478:Cells, Cultured / Q000166:cytology* / Q000378:metabolism*; D003199:Computer Systems; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000295:instrumentation*; D018613:Microscopy, Confocal / Q000295:instrumentation*; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation*; D008858:Microscopy, Phase-Contrast / Q000295:instrumentation*; D057054:Molecular Imaging / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018511:Systems Integration