Standing-wave excitation for fluorescence microscopy

Lanni, Frederick; Bailey, Brent

doi:10.1016/0962-8924(94)90125-2

Cited by 19 publications

(22 citation statements)

References 13 publications

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“…This can be achieved by leaving the objects to be observed stationary in the focal plane of the objective lens and tracking the objects_ trajectories in the standing-wave field instead of sampling it (Lanni et al 1986). With such a procedure, diffusion parameters of small objects might be accessible with a precision in the nanometer range.…”

Section: Discussionmentioning

confidence: 99%

“…To generate the illumination pattern, two counterpropagating laser beams are brought to constructive interference, establishing a standing-wave field (Lanni et al 1986, Bailey et al 1993. In contrast to focused laser light techniques (Hell et al 1994, Klar et al 2000, Dyba & Hell 2002, Egner et al 2002 or structured illumination (Gustafsson et al 1995, Frohn et al 2000, the optical resolution itself is not enhanced.…”

Section: Introductionmentioning

confidence: 99%

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High-precision structural analysis of subnuclear complexes in fixed and live cells via spatially modulated illumination (SMI) microscopy

et al. 2008

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Spatially modulated illumination (SMI) microscopy is a method of wide field fluorescence microscopy featuring interferometric illumination, which delivers structural information about nanoscale architecture in fluorescently labelled cells. The first prototype of the SMI microscope proved its applicability to a wide range of biological questions. For the SMI live cell imaging this system was enhanced in terms of the development of a completely new upright configuration. This so called Vertico-SMI transfers the advantages of SMI nanoscaling to vital biological systems, and is shown to work consistently at different temperatures using both oil-and waterimmersion objective lenses. Furthermore, we increased the speed of data acquisition to minimize errors in the detection signal resulting from cellular or object movement. By performing accurate characterization, the present Vertico-SMI now offers a fully-fledged microscope enabling a complete three-dimensional (3D) SMI data stack to be acquired in less than 2 seconds. We have performed live cell measurements of a tet-operator repeat insert in U2OS cells, which provided the first in vivo signatures of subnuclear complexes. Furthermore, we have successfully implemented an optional optical configuration allowing the generation of high-resolution localization microscopy images of a nuclear pore complex distribution. Abbreviations

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

High-precision structural analysis of subnuclear complexes in fixed and live cells via spatially modulated illumination (SMI) microscopy

et al. 2008

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“…Photographic or electronic imaging (Brinkley and Cox 1978;Cherry 1985;Waggoner et al 1989) and different fluorescent labels with multiple filter combinations (Small et al 1988;Koller 1989;Haugland 1996;Juarranz et al 1996) are currently used for multiple protein localizations with conventional, confocal, stereo, or standing-wave fluorescence microscopy (Osborn et al 1978;Lanni 1986;Ploem 1986). Several approaches have been used for the visualization and analysis of different cell structures after multiple labeling.…”

Section: Discussionmentioning

confidence: 99%

Recycling cultured cells for immunofluorescent labeling

Espada

Juarranz

Villanueva

et al. 2001

Histochem Cell Biol

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A method to use sequential rounds of immunofluorescent labeling in cell cultures is presented. The method is based on the utilization of a non-liquid reducing agent, sodium dithionite, in conjunction with ionic or non-ionic detergents (SDS or TX100, respectively) at room temperature. This method preserves cell morphology and substrate antigenicity, and operates through the complete extraction of most primary and secondary antibodies. Using this protocol, the sequential immunolocalization of different proteins is possible, without signal interference with previous immunolabeling rounds. In addition, the method is also useful to recycle blotted membranes in immunoblots.

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“…Interference set up in the excitation or fluorescence emission fields can lead to an expansion of the 0Th In DI systems for standing-wave fluorescence microscopy (SWFM), the use of an axially-periodic interference pattern (standing waves) in the excitation field generates two axial sidebands in the OTF that pass additional information into the image field ( Figure 4) (35)(36)(37)(38)(39)(40). High-contrast discrimination of labeled actin filaments separated axially by <0.3 tim, and contour tracing of cytoskeletal filaments in increments as small as 0.036 pm have been demonstrated by this method.…”

Section: Optical Systems For Enhanced 3d Resolutionmentioning

confidence: 99%

<title>Three-dimensional fluorescence microscopy of cells</title>

et al. 1996

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Applications of fluorescence microscopy in biology range from macromolecular to multicellular, with living specimens in particular placing severe constraints on the resolution, precision, speed, and optical efficiency of the imageforming instrumentation. In most cases, a high-resolution optical system will produce images in which the axial position of a feature in the specimen is encoded through sharpness of focus, leading to inherently three-dimensional (3D) data. Even in the case where quantitative analysis of only one particular plane of focus within the specimen is to be made, fluorescence from that stratum must be isolated from out-of-focus contributions. At the present time, (i) confocal scanning and (ii) direct imaging used with computational deconvolution are the main methods by which object structure is extracted from serial-focus fluorescence image sets. In addition, fluorescence microscopes utilizing interferometric illumination, multi-photon excitation, composite apertures and image interferometry have been introduced specifically to improve 3D resolution. In this report, we look specifically at the constraints imposed by biological specimens on instrumentation used for 3D fluorescence imaging.

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Standing-wave excitation for fluorescence microscopy

Cited by 19 publications

References 13 publications

High-precision structural analysis of subnuclear complexes in fixed and live cells via spatially modulated illumination (SMI) microscopy

High-precision structural analysis of subnuclear complexes in fixed and live cells via spatially modulated illumination (SMI) microscopy

Recycling cultured cells for immunofluorescent labeling

<title>Three-dimensional fluorescence microscopy of cells</title>

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