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  • 1
    Keywords: CELL ; Germany ; MICROSCOPY ; imaging ; SAMPLES ; RESOLUTION ; MARKER ; MARKERS ; ENDOPLASMIC-RETICULUM ; methods ; STIMULATED-EMISSION ; DEPLETION ; subdiffraction ; FLUORESCENT PROTEIN
    Abstract: We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy with GFP-labeled samples. The similar to 70 nm lateral resolution attained in this study is demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell. Our results mark the advent of nanoscale biological microscopy with genetically encoded markers
    Type of Publication: Journal article published
    PubMed ID: 16896340
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  • 2
    Keywords: CELLS ; CELL ; Germany ; MICROSCOPY ; imaging ; SUPPORT ; SYSTEMS ; SAMPLE ; SAMPLES ; RESOLUTION ; SUFFICIENT ; FREQUENCY ; FIELD ; FREQUENCIES ; DISPLAY ; REGION ; LIVE CELLS ; FLUORESCENCE ; max ; SECTIONS ; LIGHT ; 2-PHOTON EXCITATION ; 3-DIMENSIONAL TRANSFER-FUNCTIONS ; AXIAL RESOLUTION INCREASE ; COHERENT USE ; CONFOCAL MICROSCOPES ; ELECTROMAGNETIC DIFFRACTION ; FLUORESCENCE MICROSCOPY ; IMAGE-RESTORATION ; OPPOSING LENSES ; three-dimensional imaging
    Abstract: Although the addition of just the excitation light field at the focus, or of just the fluorescence field at the detector is sufficient for a three-to fivefold resolution increase in 4Pi-.uorescence microscopy, substantial improvements of its optical properties are achieved by exploiting both effects simultaneously. They encompass not only an additional expansion of the optical bandwidth, but also an amplified transfer of the newly gained spatial frequencies to the image. Here we report on the realization and the imaging properties of this 4Pi microscopy mode of type C that also is the far-field microscope with the hitherto largest aperture. We show that in conjunction with two-photon excitation, the resulting optical transfer function displays a sevenfold improvement of axial three-dimensional resolution over confocal microscopy in aqueous samples, and more importantly, a marked transfer of all frequencies within its inner region of support. The latter is present also without the confocal pinhole. Thus, linear image deconvolution is possible both for confocalized and nonconfocalized live-cell 4Pi imaging. Realized in a state-of-the-art scanning microscope, this approach enables robust three-dimensional imaging of fixed and live cells at; 80 nm axial resolution
    Type of Publication: Journal article published
    PubMed ID: 15377532
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