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Immersion Freezing for Cryo-Transmission Electron Microscopy: Fundamentals

The high vacuum required in a transmission electron microscope (TEM) greatly impairs the ability to study specimens naturally occurring in an aqueous phase: exposing "wet" specimens to a pressure…

Video Tutorials: Filling and Assembling of Different Carriers for High-Pressure Freezing

High pressure freezing (HPF) is a cryo-fixation method primarily for biological samples, but also for a variety of non-biological materials. It is a technique that yields optimal preservation in many…

Brief Introduction to Freeze Fracture and Etching

Freeze fracture describes the technique of breaking a frozen specimen to reveal internal structures. Freeze etching is the sublimation of surface ice under vacuum to reveal details of the fractured…

Brief Introduction to Specimen Trimming

Before ultrathin sectioning a sample with an ultramicrotome it has to be pre-prepared. For this pre-preparation, special attention must be paid to the sample size (size of the section), location of…

Glycerol Spraying/Platinum Low Angle Rotary Shadowing of DNA with the Leica EM ACE600 e-beam

Glycerol spraying/low angle rotary shadowing (Aebi and Baschong, 2006) is a preparation technique used in biology to visualize structures yielding insufficent contrast with other techniques, due to…

Brief Introduction to Freeze Substitution

Freeze-substitution is a process of dehydration, performed at temperatures low enough to avoid the formation of ice crystals and to circumvent the damaging effects observed after ambient-temperature…

Thermodynamic Considerations Regarding the LN2 in a High Pressure Freezer

Employing liquid nitrogen (LN2) as a coolant in the complex process of high pressure freezing raises certain considerations regarding phase transition not only of the liquid sample to be frozen but…

Brief Introduction to Contrasting for EM Sample Preparation

Since the contrast in the electron microscope depends primarily on the differences in the electron density of the organic molecules in the cell, the efficiency of a stain is determined by the atomic…

Brief Introduction to Glass Knifemaking for Electron and Light Microscope Applications

Glass knives are used in an ultramicrotome to cut ultrathin slices of samples for electron and light microscope applications. For resin and for cryo sections (Tokuyasu samples) the knife edge must be…