A5010878454- Advanced Fluorescence Microscopy Techniques
- Near-Field Optical Microscopy
- Advanced Electron Microscopy Techniques and Applications
- Image Processing Techniques and Applications
- Cell Image Analysis Techniques
- Digital Holography and Microscopy
- Optical Coherence Tomography Applications
- Anatomy and Medical Technology
- Force Microscopy Techniques and Applications
- Scoliosis diagnosis and treatment
- Cellular Mechanics and Interactions
- Hip disorders and treatments
University of Glasgow
2019-2021
The spatial resolution of conventional optical microscopy is limited by diffraction to transverse and axial resolutions about 250 nm, but localization point sources, such as single molecules or fluorescent beads, can be achieved with a precision 10 nm better in each direction. Traditional approaches two dimensions enable high only for thin in-focus layer that typically much less than the depth cell. This precludes, example, super-resolution extended three-dimensional biological structures...
We demonstrate how a Twin-Airy Point Spread Function can be employed in super-resolution microscopy to combine state-of-the-art 3D localisation precision and depth of field with the potential for an order-of-magnitude increase emitter densities.
Localisation microscopy provides a dramatic resolution enhancement over conventional microscopy, capable of imaging biological structures at the scale individual molecules. The image-acquisition speed is however limited by requirement to time-sequentially image fluorescence emissions without significant overlap emitter images. For 3D localisation each emission encoded with spatially extended point-spread function, which further reduces maximum usable density emitters in acquired images, and...
Localisation microscopy provides a dramatic resolution enhancement over conventional microscopy, capable of imaging biological structures at the scale individual molecules. The image-acquisition speed is however limited by requirement to time-sequentially image fluorescence emissions without significant overlap emitter images. For 3D localisation each emission encoded with spatially extended point-spread function, which further reduces maximum usable density emitters in acquired images, and...
Precise 3D point localization is increasingly important in microscopy, but algorithms break down when PSFs overlap. We adapt the CLEAN algorithm from astronomical imaging to enable MLE of high-density datasets.
Computational imaging with engineered pupil functions has been plagued by image artefacts. We discuss solutions to this problem that yield snapshot 3D and localisation microscopy the highest reported voxel count.
Measurement of cellular tractions is important for, for example, engineering artificial tissue. We propose the use PSF-engineered localisation microscopy to achieve video rate measurements in large volumes with minimal sample light exposure.