Correlated atomic force microscopy and fluorescence lifetime imaging of live bacterial cells Export

@article{citeulike:4571133, abstract = {We report on imaging living bacterial cells by using a correlated tapping-mode atomic force microscopy (AFM) and confocal fluorescence lifetime imaging microscopy (FLIM). For optimal imaging of Gram-negative Shewanella oneidensis MR-1 cells, we explored different methods of bacterial sample preparation, such as spreading the cells on poly- -lysine coated surfaces or agarose gel coated surfaces. We have found that the agarose gel containing 99\% ammonium acetate buffer can provide sufficient local aqueous environment for single bacterial cells. Furthermore, the cell surface topography can be characterized by tapping-mode in-air AFM imaging for the single bacterial cells that are partially embedded. Using in-air rather than under-water AFM imaging of the living cells significantly enhanced the contrast and signal-to-noise ratio of the AFM images. Near-field AFM-tip-enhanced fluorescence lifetime imaging (AFM–FLIM) holds high promise on obtaining fluorescence images beyond optical diffraction limited spatial resolution. We have previously demonstrated near-field AFM–FLIM imaging of polymer beads beyond diffraction limited spatial resolution. Here, as the first step of applying AFM–FLIM on imaging bacterial living cells, we demonstrated a correlated and consecutive AFM topographic imaging, fluorescence intensity imaging, and FLIM imaging of living bacterial cells to characterize cell polarity.}, author = {Micic, M.}, citeulike-article-id = {4571133}, citeulike-linkout-0 = {http://dx.doi.org/10.1016/j.colsurfb.2003.10.020}, citeulike-linkout-1 = {http://linkinghub.elsevier.com/retrieve/pii/S0927-7765(03)00278-9}, day = {15}, doi = {10.1016/j.colsurfb.2003.10.020}, issn = {09277765}, journal = {Colloids and Surfaces B: Biointerfaces}, keywords = {afm, live, oneidensis, shewanella, thesis}, month = {April}, number = {4}, pages = {205--212}, posted-at = {2009-05-21 17:39:32}, priority = {2}, title = {Correlated atomic force microscopy and fluorescence lifetime imaging of live bacterial cells}, url = {http://dx.doi.org/10.1016/j.colsurfb.2003.10.020}, volume = {34}, year = {2004} }

TY - JOUR ID - citeulike:4571133 L3 - citeulike-article-id:4571133 N2 - We report on imaging living bacterial cells by using a correlated tapping-mode atomic force microscopy (AFM) and confocal fluorescence lifetime imaging microscopy (FLIM). For optimal imaging of Gram-negative Shewanella oneidensis MR-1 cells, we explored different methods of bacterial sample preparation, such as spreading the cells on poly- -lysine coated surfaces or agarose gel coated surfaces. We have found that the agarose gel containing 99% ammonium acetate buffer can provide sufficient local aqueous environment for single bacterial cells. Furthermore, the cell surface topography can be characterized by tapping-mode in-air AFM imaging for the single bacterial cells that are partially embedded. Using in-air rather than under-water AFM imaging of the living cells significantly enhanced the contrast and signal-to-noise ratio of the AFM images. Near-field AFM-tip-enhanced fluorescence lifetime imaging (AFM–FLIM) holds high promise on obtaining fluorescence images beyond optical diffraction limited spatial resolution. We have previously demonstrated near-field AFM–FLIM imaging of polymer beads beyond diffraction limited spatial resolution. Here, as the first step of applying AFM–FLIM on imaging bacterial living cells, we demonstrated a correlated and consecutive AFM topographic imaging, fluorescence intensity imaging, and FLIM imaging of living bacterial cells to characterize cell polarity. IS - 4 JF - Colloids and Surfaces B: Biointerfaces SN - 09277765 EP - 212 TI - Correlated atomic force microscopy and fluorescence lifetime imaging of live bacterial cells VL - 34 SP - 205 KW - afm KW - live KW - oneidensis KW - shewanella KW - thesis AU - Micic, M PY - 2004/04/15/ UR - http://dx.doi.org/10.1016/j.colsurfb.2003.10.020 ER -