Methods

To be able to view a biological sample in the electron microscope it must first be stabilized or "fixed", preferrably in a way that the ultrastructure of the cells or tissue remain as close to the living material as possible. The choice of fixative...

The routine method for examining biological samples in TEM is to embed the material in plastic and cut ultrathin (60-80nm) sections. The choice of resin depends on the type of sample and the purpose of your study. Listed below are some of the more...

Preparation of ultrathin frozen sections (Tokuyasu) Fixation of cells in culture: Attached cells: first remove cells from the dish with 5mM EDTA/PBS using a transfer pipet with a wide opening. (You can use trypsin if you are not interested in surface...

5µl of the sample is adsorbed for 1 minute to a carbon coated grid that has been made hydrophilic by a 20 second exposure to a glow discharge (25mA). Excess liquid is removed with a filterpaper (Whatman #1), the grid is then floated briefly on a drop of...

Fixation Fixation 1-2 hrs (or overnight at 4C) 2.5% Glutaraldehyde 2.5% Paraformaldehyde in 0.1 M sodium cacodylate buffer (pH 7.4). Rinse 3 X 10 min in 0.1M sodium cacodylate buffer, pH 7.4 Post-Fix 1-2 hrs in 1% Osmium tetroxide in water Rinse 3 X 5 min...