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Anatomical Methods
1. Videos from 2023 International C. elegans Conference Electron Microscopy Workshop (click on title for full size video)
Application of high-pressure freeze serial TEM for single-cell reconstruction and immunohistochemistry by Mia Krout
vEM for worm connectomics and more by Ben Mulcahy
How hard is it to do EM on worms? Sample prep tips for conventional and more fancy EM (FIB-SEM) by Sebastian Markert
Array tomography: How to democratize C. elegans EM by Irina Kolotuev
2. Fixation and Embedding for TEM
Immersion Fixation Protocol for Structure
Microwave-Assisted Fixation for Ultrastructure
Microwave-Assisted Fixation for Immunocytochemistry
High Pressure Freeze (HPF) and Freeze Substitution
Pre-Embedding Antibody Staining for TEM
Older Methods
Conventional two-step fixation by David H. Hall
Freeze substitution with tannic acid/osmium_long incubation by Robby Weimer - pdf file
Freeze substitution with tannic acid/osmium_short incubation by Robby Weimer - pdf file
High pressure freezing/freeze-substitution of C. elegans embryos and L1 worms by Richard Fetter
Freeze substitution in Potassium Permanganate by Robby Weimer - pdf file
Metal mirror (slam-freeze) fixation (MMF) by David H. Hall
Flat embedding of worms (slam freezing) by Steve Fields (Rand Lab)
Laser hole fixation of embryos by Carolyn Norris (Hedgecock Lab)
3. Fixation and Embedding for SEM
Fixation for Block-Face Imaging, ATUM & FIB/SEM (OTO)
SEM preparation of dissected worms by David Greenstein
4. Sectioning
Serial Thin Sectioning - The Hartwieg Method
Cryo-Thin Sections
Post-Embedding Antibody Staining for TEM
Alternate Methods for Post-Staining Thin Sections
Grid Making
ATUM (automated) Thin Sections
5. Imaging (TEM and SEM)
Serial Block-Face SEM
FIB/SEM
Viewing ATUM Sections on Tape
Electron Tomography Imaging
6. Staining for Light and Fluorescence Microscopy
Mounting animals for observation with Nomarski DIC Optics by Monica Driscoll
Freeze-crack and staining by Janet Duerr
FMRFamide staining by Chris Li
Gonad-intestine staining by Barth Grant
Antibody staining of formaldehyde-fixed animals by Gary Ruvkun and Michael Finney
Staining of early embryos by Bruce Bowerman
Formaldehyde fixation and cytoskeletal staining by Raffi Aroian
DiI and DiO staining methods
SYTO staining of nucleic acids in live worms by Hill and L'Hernault
DAPI (4'-6-Diamidino-2-phenylindole) staining of nuclei by Ross Francis (modified by Sudhir Nayak)
FITC (fluorescein isothiocyanate) staining of amphids and phasmids by Ed Hedgecock
7. Image Analysis and Serial Reconstruction
Electron Tomography
Computer-Based Analytical Tools
8. See Electron Microscopy Methods in Cell Biology in WormBook
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