Darkfield Imaging
Darkfield Imaging - 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.
13:45
Hybrid µCT-FMT imaging and image analysis
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Cited by 34 •
2015
We describe a protocol for hybrid imaging, combining fluorescence-mediated tomography (FMT) with micro computed tomography (µCT). After fusion and reconstruction, we perform interactive organ segmentation to extract quantitative measurements of the fluorescence distribution.
14:09
Localization and Relative Quantification of Carbon Nanotubes in Cells with Multispectral Imaging Flow Cytometry
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Cited by 18 •
2013
In this study, the main purpose was to monitor the cellular uptake and eventual exocytosis of carbon nanotubes (CNTs). From this perspective, we proposed here a unique method using multispectral imaging flow cytometry allowing a quantification and localization of CNTs in a statistically relevant number of cells.
FAQs
The image contrast arises from separating illumination from detection. Oblique light reaches the specimen, but directly transmitted light is blocked before it can enter the objective. Cells, particles, and surface features redirect part of that illumination through scattering, allowing the objective to collect those signals and render the corresponding structures against the dark field.
Visibility depends on whether a specimen contains features that redirect the oblique illumination. Cellular structures, microorganisms, particles, and surface features can produce detectable scattered light even when their natural contrast is limited. Consequently, the technique can reveal morphology or particle behavior without requiring a fluorescent label or staining step.
Darkfield Imaging provides a label-free alternative when researchers want to preserve specimens or avoid fluorescent labels. Brightfield and fluorescence microscopy offer different contrast strategies, whereas darkfield detection emphasizes light scattered by the sample. Using these approaches as complements can help researchers select imaging conditions suited to morphology, dynamic behavior, or sample preservation.
A functional arrangement must create oblique illumination, prevent directly transmitted light from reaching the objective, and allow scattered light from the specimen to enter the objective. This sequence is essential: without the illumination angle, relevant scattering may not be generated, and without the light-blocking step, the background would not remain dark enough to emphasize the specimen.
The method is useful for observing cell morphology, microorganism motility, particle behavior, and dynamic interactions in engineered biological systems. These applications benefit from viewing specimens without staining, particularly when labeling could interfere with preservation goals or when researchers need to monitor changes and interactions over time in a noninvasive manner.
Darkfield observations can provide visual information about specimen shape, movement, particle behavior, and interactions among components of an engineered biological system. Because the approach avoids staining and fluorescent labels, it supports noninvasive monitoring while preserving the sample. The resulting images can complement other microscopy data when structural or dynamic behavior is the main focus.