Darkfield Microscopy
Darkfield Microscopy - Related Videos
05:15
Darkfield Microscopy to Visualize Diffusional Dynamics of Nanorods on Cell Membrane
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2025
In this video, we describe the darkfield microscopy (DFM) technique combined with single-particle tracking (SPT) analysis to monitor the interaction of plasmonic gold nanorods with the cell membrane of a specific cell line of interest. Combined DFM and SPT analysis allows the visualization of the nanorod movement through the cell membrane with high spatiotemporal resolution.
12:19
Identification of Metal Oxide Nanoparticles in Histological Samples by Enhanced Darkfield Microscopy and Hyperspectral Mapping
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Cited by 37 •
2015
Enhanced darkfield microscopy and hyperspectral imaging with spectral mapping enable screening, localization, and identification of nanoscale materials in histological samples with improved speed and accuracy over traditional methods. The goal of this paper is to provide methods for darkfield imaging and hyperspectral mapping of metal oxide nanoparticles in histological samples.
09:09
Visualizing Diffusional Dynamics of Gold Nanorods on Cell Membrane using Single Nanoparticle Darkfield Microscopy
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Cited by 1 •
2021
Here, we show the use of traditional dark-field microscopy to monitor the dynamics of gold nanorods (AuNRs) on cell membrane. The location and orientation of single AuNRs are detected using ImageJ and MATLAB, and the diffusive states of AuNRs are characterized by single particle tracking analysis.
10:00
Low-Cost Cryo-Light Microscopy Stage Fabrication for Correlated Light/Electron Microscopy
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Cited by 11 •
2011
We demonstrate the fabrication of a low-cost cryogenic stage designed to fit most reflected light microscopes. This lab-built cryogenic stage enables efficient and reliable correlative imaging between cryo-light and cryo-electron microscopy.
11:16
Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
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Cited by 12 •
2016
A method is described whereby quantum dot (QD) nanoparticles can be used for correlative immunocytochemical studies of epoxy embedded human pathology tissue. We employ commercial antibody fragment conjugated QDs that are visualized by widefield fluorescence light microscopy and transmission electron microscopy.
FAQs
Contrast arises because a central opaque stop blocks direct transmitted light before it reaches the specimen’s image-forming path. A specialized condenser instead directs illumination at an oblique angle. Transparent or lightly scattering cells and structures redirect part of that light into the objective, making those features visible against the dark field.
The opaque stop and condenser work as a paired illumination system rather than as independent accessories. The stop removes direct transmitted illumination, while the condenser establishes the oblique directions needed for scattering. Image formation therefore depends on light being redirected by the specimen; without that scattered contribution, transparent material provides little visible signal.
Darkfield Microscopy differs from routine brightfield viewing mainly in how contrast is produced. It can show transparent, unstained material through scattered light instead of relying on routine staining, and it may expose structures that are difficult to distinguish in brightfield images. This makes the approach especially useful when preserving live-sample observation matters.
A practical observation begins with a transparent or lightly scattering biological sample that does not require routine staining. The microscope uses the central opaque stop together with the specialized condenser to provide oblique illumination, after which the observer examines light scattered by the specimen. This workflow supports viewing cells or microorganisms while they remain unstained.
Use this method when rapid, noninvasive viewing is more important than producing a stained preparation. In biology, it is suited to live, unstained samples and can support bacterial morphology and motility observations. It also serves screening and teaching, where immediate visual contrast helps users examine specimens without routine staining.
The resulting images can provide evidence about cell or microorganism shape, visible fine structures, and movement over time. Because the sample can be observed live and without routine staining, the technique is useful for studying dynamic cellular behavior rather than only fixed appearance. Its value is strongest when motion or subtle scattering features are biologically relevant.