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Imaging cellular structures
Imaging cellular structures
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To localize cellular structures down to the molecular level fluorescence labeling is an indispensable technique. Topographic information of the corresponding structures or the cellular compartments hosting the structures is another refinement that can be provided by AFM imaging.
Here L929 fibroblasts and MC3T3 osteoblasts were stained for f-actin using AlexaFluor546-and FITC-phalloidin respectively. L929 cells were grown and fixed on glass slides, the MC3T3 cells on coverslips for the use with the CoverslipHolder™. To prevent photo bleaching, focus and adequate sample position were adjusted using fluorescence detection in binning mode. High quality images for the overlay were taken before calibration images. Applying phalloidin staining the actin stress fibers could be visualized using fluorescence microscopy. Additional topographical information could be derived by AFM imaging, also revealing the fine structure of the filaments and thus complementing the optical image.
Here L929 fibroblasts and MC3T3 osteoblasts were stained for f-actin using AlexaFluor546-and FITC-phalloidin respectively. L929 cells were grown and fixed on glass slides, the MC3T3 cells on coverslips for the use with the CoverslipHolder™. To prevent photo bleaching, focus and adequate sample position were adjusted using fluorescence detection in binning mode. High quality images for the overlay were taken before calibration images. Applying phalloidin staining the actin stress fibers could be visualized using fluorescence microscopy. Additional topographical information could be derived by AFM imaging, also revealing the fine structure of the filaments and thus complementing the optical image.