Ultramicrotomy

Ultramicrotomy is a method for cutting specimens into extremely thin slices, called ultra-thin sections, that can be studied and documented at different magnifications in a transmission electron microscope (TEM). It is used mostly for biological specimens, but sections of plastics and soft metals can also be prepared. Sections must be very thin because the 50 to 125 kV electrons of the standard electron microscope cannot pass through biological material much thicker than 150 nm. For best resolutions, sections should be from 30 to 60 nm. This is roughly the equivalent to splitting a 0.1 mm-thick human hair into 2,000 slices along its diameter, or cutting a single red blood cell into 100 slices. Ultramicrotomy is a method for cutting specimens into extremely thin slices, called ultra-thin sections, that can be studied and documented at different magnifications in a transmission electron microscope (TEM). It is used mostly for biological specimens, but sections of plastics and soft metals can also be prepared. Sections must be very thin because the 50 to 125 kV electrons of the standard electron microscope cannot pass through biological material much thicker than 150 nm. For best resolutions, sections should be from 30 to 60 nm. This is roughly the equivalent to splitting a 0.1 mm-thick human hair into 2,000 slices along its diameter, or cutting a single red blood cell into 100 slices. Ultra-thin sections of specimens are cut using a specialized instrument called an 'ultramicrotome'. The ultramicrotome is fitted with either a diamond knife, for most biological ultra-thin sectioning, or a glass knife, often used for initial cuts. There are numerous other pieces of equipment involved in the ultramicrotomy process. Before selecting an area of the specimen block to be ultra-thin sectioned, the technician examines semithin or 'thick' sections range from 0.5 to 2 µm. These thick sections are also known as survey sections and are viewed under a light microscope to determine whether the right area of the specimen is in a position for thin sectioning. 'Ultra-thin' sections from 50 to 100 nm thick are able to be viewed in the TEM.