ArticleNature methods2024
POLCAM: instant molecular orientation microscopy for the life sciences.
Article in Nature methods, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- SPIFFI enables single-shot super-resolution and multidimensional imaging.Nature methods · 2026Article
- Single Fluorogens and Orientation-Localization Microscopy for Quantifying Chemical and Biomolecular Dynamics at the Nanoscale.Accounts of chemical research · 2026Article
- Fused deep learning enables 6D single-molecule localization in polarization-resolved microscopy.Methods and applications in fluorescence · 2026Article
- 4polar3D single molecule imaging of 3D orientation in dense actin networks using ratiometric polarization splitting.Nature communications · 2026Article
- FLIPs: Genetically encoded molecular biosensors for functional imaging of cell signaling by linear dichroism microscopy.Science advances · 2026Article
- Dual-view microscopy of single-molecule dipole orientations.Biophysical reports · 2025Article
- Single-shot Stokes polarimetry of plasmon-coupled single-molecule fluorescence.Nanophotonics (Berlin, Germany) · 2025Article
- Single molecule orientation and localization microscopy.Nature photonics · 2025Article
- Advanced optical microscopy methods forBiophysical reviews · 2025Review
- Dipole Determination by Polarimetric Spectroscopy Yielding the Orientation of Gold Nanorods.Small science · 2025Article
- Single-shot full-field optical coherence tomography with a single polarization camera.Biomedical optics express · 2025Article
- Painting rich six-dimensional pictures using polarized fluorescence microscopy.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Single-molecule orientation-localization microscopy: Applications and approaches.Quarterly reviews of biophysics · 2024Review
- RASP: Optimal Single Puncta Detection in Complex Cellular Backgrounds.The journal of physical chemistry. B · 2024Article
Corrections and comments
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Authors and funding
15 authors.
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Abstract
Current methods for single-molecule orientation localization microscopy (SMOLM) require optical setups and algorithms that can be prohibitively slow and complex, limiting widespread adoption for biological applications. We present POLCAM, a simplified SMOLM method based on polarized detection using a polarization camera, which can be easily implemented on any wide-field fluorescence microscope. To make polarization cameras compatible with single-molecule detection, we developed theory to minimize field-of-view errors, used simulations to optimize experimental design and developed a fast algorithm based on Stokes parameter estimation that can operate over 1,000-fold faster than the state of the art, enabling near-instant determination of molecular anisotropy. To aid in the adoption of POLCAM, we developed open-source image analysis software and a website detailing hardware installation and software use. To illustrate the potential of POLCAM in the life sciences, we applied our method to study α-synuclein fibrils, the actin cytoskeleton of mammalian cells, fibroblast-like cells and the plasma membrane of live human T cells.
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Registered trials
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