Fluorescence Polarization Microscopy
Fluorescence Polarization Microscopy - Related Videos
09:22
Introduction to Fluorescence Microscopy
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2023
Fluorescence microscopy is a very powerful analytical tool that combines the magnifying properties of light microscopy with visualization of fluorescence. Fluorescence is a phenomenon that involves absorbance and emission of a small range of light wavelengths by a fluorescent molecule known as a fluorophore. Fluorescence microscopy is accomplished in conjunction with the basic light microscope by the addition of a powerful light source, specialized filters, and a means of fluorescently...
11:23
Lensless Fluorescent Microscopy on a Chip
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Cited by 8 •
2011
A lensless on-chip fluorescent microscopy platform is demonstrated that can image fluorescent objects over an ultra-wide field-of-view of e.g., >0.6-8 cm2 with <4μm resolution using a compressive sampling based decoding algorithm. Such a compact and wide-field fluorescent on-chip imaging modality could be valuable for high-throughput cytometry, rare-cell research and microarray-analysis.
01:01
Group Polarization
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2020
Group polarization is the strengthening of an original group attitude following the discussion of views within a group (Teger & Pruitt, 1967). That is, if a group initially favors a viewpoint, after discussion the group consensus is likely a stronger endorsement of the viewpoint. Conversely, if the group was initially opposed to a viewpoint, group discussion would likely lead to stronger opposition. The phenomenon of group polarization explains many actions taken by groups that would not...
09:02
Visualizing Bacteria in Nematodes using Fluorescent Microscopy
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Cited by 20 •
2012
To study the mutualism between Xenorhabdus bacteria and Steinernema nematodes, methods were developed to monitor bacterial presence and location within nematodes. The experimental approach, which can be applied to other systems, entails engineering bacteria to express the green fluorescent protein and visualizing, using fluorescence microscopy bacteria within the transparent nematode.
08:27
Visualizing Stromule Frequency with Fluorescence Microscopy
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Cited by 5 •
2016
Protocols to investigate the dynamics of chloroplast stromules, the stroma-filled tubules that extend from the surface of chloroplasts, are described.
FAQs
The key signal is the extent to which emitted light preserves the polarization imposed during excitation. Fluorophores that rotate rapidly during the relevant measurement interval change their orientation and produce a more depolarized signal. Restricted rotational motion or aligned molecules preserve more polarization, allowing differences in molecular mobility or organization to become measurable.
Fluorescence polarization or anisotropy adds information about orientation and rotational mobility to the spatial information obtained from fluorescence imaging. A signal can therefore indicate whether molecules are aligned, rapidly rotating, or moving more slowly, rather than simply showing where fluorescence is present. This extra dimension supports analysis of molecular organization and interactions in biological samples.
Protein binding studies benefit from this method because molecular interactions can be evaluated through their effects on the polarization readout. When fluorescent molecules participate in a different organizational or motion state, the measured fluorescence polarization or anisotropy provides a biophysical signal associated with that state. This complements imaging of where proteins appear by addressing how their organization or mobility changes.
A basic measurement begins by illuminating the sample with polarized excitation light, then analyzing the polarization of the fluorescence emitted by the labeled molecules. Researchers quantify the resulting fluorescence polarization or anisotropy and relate the value to molecular orientation or rotational mobility. In biological samples, this workflow can be applied to cells and tissues to connect optical measurements with organization and motion.
Researchers can apply the method to questions about protein binding, membrane organization, and cytoskeletal structure. Each application uses polarization behavior as a readout of how fluorescent molecules move or align within the relevant biological setting. The resulting measurements add biophysical information to studies of cellular architecture, helping connect molecular arrangement with structural organization.
In biology, cellular function depends on both where molecules are located and how they move. Polarization measurements link those dimensions by reporting molecular dynamics and spatial organization in cells and tissues. This perspective can help researchers interpret how changes in membrane, cytoskeletal, or protein-associated organization contribute to cellular function.