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Electron Microscopy Preparation of Grid

Electron Microscopy Preparation of Grid:

It is extremely difficult to handle ultrathin sections directly. They are removed by a small circular copper grid that is having 400 apertures per inch square. Even this degree of support is insufficient, therefore, it is a customary to coat the grids with a thin electron transparent layer of collodion, carbon or Formvar. Carbon support films are stronger and Formvar coating is frequently employed as a foundation film

onto which carbon is deposited, the Formvar is then dissolved away. For coating take a clean, dry microscope  slide and put. It in a coupling jar having 0.5% Formvar solution in chloroform. After five seconds with raw the slide smoothly from the coupling jar and allow it to dry. Cut the film around the edges. Breath heavily on the film. Float off the Formvar membrane by lowering th slide at a shallow angle into a bowl of clean water filled upto the rim. Place clean grids on the floating film by keeping dull side downwards. Using a flat surface of clean microscope slide, push he film down into the water. Smoothly and quickly turn the slide in such a way that the film is lifted out. The film can also be fished out from water by simply lowering the Parafilm held  horizontally without pushing the film in the water. Dry the film.

The evaporated carbon films are prepared by deposition of thin carbon layers (5-10nm) under vacuum over the Formvar film (19). Generally, Formvar is removed from the grid if they are to be used for sections. Transfer carefully for Formvar and carbon coated grids from slide to a fine copper wiremesh submerged chloroform. This will dissolve away the Formvar, leaving the carbon film intact. Carefully remove chloroform from the container with pipette. Remove the gird and place on a filter paper in a petri dish.

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