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Related Concept Videos

Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
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Fixation and Sectioning

Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
Simple Staining Technique01:24

Simple Staining Technique

OverviewStaining techniques in microscopy enhance the visualization of microorganisms by increasing contrast and allowing the differentiation of cellular structures. Simple staining is one of the fundamental methods used to observe the basic morphological characteristics of microorganisms, including their size, shape, and arrangement. This method relies on the application of a single dye to stain the entire cell, producing a clear contrast between the cell and the background.FixationFixation is...

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Microscopy Techniques for Interpreting Fungal Colonization in Mycoheterotrophic Plants Tissues and Symbiotic Germination of Seeds
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Sample preparation for STED microscopy.

Christian A Wurm1, Daniel Neumann, Roman Schmidt

  • 1Mitochondrial Structure and Dynamics/Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Goettingen, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|December 4, 2009
PubMed
Summary
This summary is machine-generated.

Researchers developed sample preparation protocols for Stimulated Emission Depletion (STED) microscopy. These methods enable sub-diffraction resolution imaging for fluorescence microscopy, overcoming traditional optical limits.

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Area of Science:

  • Biophysics
  • Optical Microscopy
  • Cell Biology

Background:

  • The diffraction limit restricts far-field optical microscopy resolution to approximately half the wavelength of light.
  • Stimulated Emission Depletion (STED) microscopy offers a way to overcome these resolution limits in fluorescence imaging.
  • Conventional sample preparation for confocal microscopy may not suffice for high-resolution STED imaging.

Purpose of the Study:

  • To present robust protocols for preparing test samples specifically for STED microscopy.
  • To provide adaptable starting points for existing labeling strategies to meet STED microscopy requirements.
  • To facilitate sub-diffraction resolution imaging in biological samples.

Main Methods:

  • Development of standardized protocols for preparing bead samples.
  • Establishment of protocols for immunolabeled mammalian cells suitable for STED microscopy.
  • Focus on ensuring structural preservation of specimens under high-resolution conditions.

Main Results:

  • Demonstrated robust protocols for generating test samples for STED microscopy.
  • Provided methods applicable to both bead samples and biological specimens (mammalian cells).
  • Enabled adaptation of existing labeling techniques for sub-diffraction resolution.

Conclusions:

  • The presented protocols are effective for preparing samples for STED microscopy.
  • These methods support the adaptation of current fluorescence imaging techniques for enhanced resolution.
  • Facilitates the use of STED microscopy for detailed structural analysis beyond the diffraction limit.