Embedding in Epoxy Resins for Ultrathin Sectioning in Electron Microscopy
Stain Technology · 1960 · 3.3K citations · 8 references
Materials ScienceTissue EngineeringEngineeringElectron MicroscopyMicroscopyMicroscopy MethodApplied PhysicsGlass KnivesElectron MicroscopeHardener DdsaBiomedical EngineeringFixed TissueUltrastructure
Araldite is manufactured in the U.S.A. and England, with differing formulations. Tissue is dehydrated in tert‑butyl alcohol, cleared in toluene, infiltrated and embedded in an Araldite resin‑hardener‑accelerator mixture (without dibutyl phthalate) polymerized at 60 °C, producing blocks that resist fracture and yield 1 µm phase‑contrast sections and ultrathin sections that cut easily with glass knives; the method includes specific proportions for U.S.
Fixed tissue is dehydrated with tertiary butyl alcohol overnight. The following day it is cleared in toluene, infiltrated and embedded in Araldite resin-hardener-accelerator mixture without dibutyl phthalate, and polymerized at 60° C. More rapid than previous techniques, this method gives blocks which do not fracture unduly on trimming and provides sections of soft tissues at 1 μ for phase contrast microscopy, as well as ultrathin sections which cut as easily with glass knives as sections of methacrylate. Araldite manufactured in the U.S.A. and in England are different. Satisfactory proportions for the American are: hardener DDSA, 3.5 ml; casting resin 6005, 5.0 ml; accelerator B, 0.12 ml. For the British product, these are: hardener 964 B, 5.0 ml; casting resin M, 5.0 ml; accelerator 964 C, 0.25 ml. The use of 2% agar for orienting small specimens in Araldite is feasible. Mallory's borax-methylene blue has been applied to the staining of Araldite sections as thin as 0.5 μ mounted on glass slides.
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Araldite as an Embedding Medium for Electron Microscopy
Audrey M. Glauert, R. H. Glauert · The Journal of Cell Biology · 1958
628 citations
A New Embedding Medium for Electron Microscopy
Audrey M. Glauert, G.E. Rogers, R. H. Glauert · Nature · 1956
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New Sectioning Techniques for Light and Electron Microscopy
Sanford B. Newman, Emil Borysko, Max Swerdlow · Science · 1949
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G.E. Rogers · Journal of Ultrastructure Research · 1959
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The Use of Potassium Permanganate as an Electron-Dense Stain for Sections of Tissue Embedded in Epoxy Resin
A. M. Lawn · The Journal of Cell Biology · 1960
198 citations