A5077950179- Advanced Electron Microscopy Techniques and Applications
- Electron and X-Ray Spectroscopy Techniques
- Ion-surface interactions and analysis
- Integrated Circuits and Semiconductor Failure Analysis
- Advanced X-ray Imaging Techniques
- Advanced Materials Characterization Techniques
- Cell Image Analysis Techniques
- Advanced Fluorescence Microscopy Techniques
- Digital Holography and Microscopy
Rosalind Franklin Institute
2022-2024
Abstract Structural biology studies inside cells and tissues require methods to thin vitrified specimens electron transparency. Until now, focused ion beams based on gallium have been used. However, implantation, changes surface chemistry an inability access high currents limit application. Here, we show that plasma-coupled sources can produce cryogenic lamellae of human in a robust automated manner, with quality sufficient for pseudo-atomic structure determination. Lamellae were produced...
Serial focussed ion beam scanning electron microscopy (FIB/SEM) enables imaging and assessment of subcellular structures on the mesoscale (10 nm to 10 µm). When applied vitrified samples, serial FIB/SEM is also a means target specific in cells tissues while maintaining constituents’ hydration shells for situ structural biology downstream. However, application non-stained cryogenic biological samples limited due low contrast, curtaining, charging artefacts. We address these challenges using...
Abstract An emergent volume electron microscopy technique called cryogenic serial plasma focused ion beam milling scanning (pFIB/SEM) can decipher complex biological structures by building a three-dimensional picture of samples at mesoscale resolution. This is achieved collecting consecutive SEM images after successive rounds FIB that expose new surface each step. Due to instrumental limitations, some image processing necessary before 3D visualization and analysis the data possible. are...
Abstract Structural biology inside cells and tissues requires methods able to thin vitrified specimens electron transparent thicknesses. Until now, focused ions beams based on gallium have been used. However, ion implantation, changes surface chemistry an inability access high currents limit Gallium as beam source. Here, we show that plasma-coupled sources can produce cryogenic lamella of human in a robust automated manner, with quality sufficient for pseudo-atomic structure determination....
Journal Article Computing for Optimized Biological Microscopy Data Processing and Analysis at The Rosalind Franklin Institute Get access Luís M A Perdigão, Perdigão Institute, Harwell Campus, Didcot, United Kingdom Corresponding author: luis.perdigao@rfi.ac.uk Search other works by this author on: Oxford Academic Google Scholar Neville B-y Yee, Yee Elaine L Ho, Ho Avery H Pennington, Pennington Diamond Light Source, Oliver NF King, King Michele C Darrow, Darrow Mark Basham Microanalysis,...
Abstract Serial focussed ion beam scanning electron microscopy (FIB/SEM) enables imaging and assessment of sub-cellular structures on the mesoscale (10 nm to 10 μm). When applied vitrified samples, serial FIB/SEM is also a means target specific in cells tissues while maintaining constituents’ hydration shells for in-situ structural biology downstream. However, application non-stained cryogenic biological samples limited due low contrast, curtaining charging artefacts. We address these...
The experimental limitations with optics observed in many microscopy and astronomy instruments result detrimental effects for the imaging of objects. This can be generally described mathematically as a convolution real object image point spread function that characterizes optical system. popular Richardson-Lucy (RL) deconvolution algorithm is widely used inverse process restoring data without these aberrations, often critical step processing data. Here we present versatile RedLionfish python...
Electron cryo-tomography is an imaging technique for probing 3D structures with at the nanometer scale. This has been used extensively in biomedical field to study complex of proteins and other macromolecules. With advancement technology, microscopes are currently capable producing images amounting terabytes data per day, posing great challenges scientists as speed processing cannot keep up ever-higher throughput microscopes. Therefore, automation essential natural pathway on which image...
Abstract An emergent volume electron microscopy (vEM) technique called cryogenic serial plasma focused ion beam milling scanning (pFIB/SEM) can decipher complex biological structures by building a three-dimensional picture of samples at mesoscale resolution. This is achieved collecting consecutive SEM images after successive rounds FIB that expose new surface each step. Due to instrumental limitations, some image processing necessary before 3D visualisation and analysis the data possible....
Abstract Electron cryo-tomography (cryo-ET) is an imaging technique for probing 3D structures with at the nanometre scale. This has been used extensively in biomedical field to study complex of proteins and other macromolecules. With advancement technology, microscopes are currently capable producing images amounting terabytes data per day, posing great challenges scientists as speed processing cannot keep up ever-higher throughput microscopes. Therefore, automation essential natural pathway...