Of all the commercially available scanning electron microscopes which operate at “low vacuum” the ESEM is the most suitable for examining biological samples. in order to maintain samples with liquid water present the specimen chamber must be capable of operating at a pressure of at least 4.6 Torr (611 pascals) of water vapour pressure (the vapour pressure of water at 0°C). Use of lower pressures or a chamber gas other than water vapour will result in evaporation of water from the sample at a rate dependant on the partial pressure difference between the sample and its surrounding environment. Tables of relative humidity as a function of water vapour pressure and temperature are readily available to calculate desired settings for the microscope.

One of the difficulties associated with examining fresh biological material is the need to have the microscope and sample available in the same location at the same time.If sample collection occurs at a site remote from the microscope inevitable necrotic changes will occur before examination can be carried out.