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• W2892384900 abstract "Abstract Correlative light and electron microscopy (CLEM) has been in use for several years, however it has remained a costly method with difficult sample preparation. Here, we report a series of technical improvements developed for precise and cost-effective correlative light and scanning electron microscopy (SEM) and focused ion beam (FIB)/SEM microscopy of single cells, as well as large tissue sections. Customized coordinate systems for both slides and coverslips were established for thin and ultra-thin embedding of a wide range of biological specimens. Immobilization of biological samples was examined with a variety of adhesives. For histological sections, a filter system for flat embedding was developed. We validated ultra-thin embedding on laser marked slides for efficient, high-resolution CLEM. Target cells can be re-located within minutes in SEM without protracted searching and correlative investigations were reduced to a minimum of preparation steps, while still reaching highest resolution. The FIB/SEM milling procedure is facilitated and significantly accelerated as: (i) milling a ramp becomes needless, (ii) significant re-deposition of milled material does not occur; and (iii) charging effects are markedly reduced. By optimizing all technical parameters FIB/SEM stacks with 2 nm iso-voxels were achieved over thousands of sections, in a wide range of biological samples." @default.
• W2892384900 created "2018-10-05" @default.
• W2892384900 creator A5040083768 @default.
• W2892384900 creator A5050402857 @default.
• W2892384900 date "2018-09-24" @default.
• W2892384900 modified "2023-10-16" @default.
• W2892384900 title "From Light Microscopy to Analytical Scanning Electron Microscopy (SEM) and Focused Ion Beam (FIB)/SEM in Biology: Fixed Coordinates, Flat Embedding, Absolute References" @default.
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• W2892384900 doi "https://doi.org/10.1017/s1431927618015015" @default.
• W2892384900 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6378657" @default.
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