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Electron Microscopy tissue sampling

Electron microscopy is sensitive to tissue handling procedures. Whilst we recognise and appreciate that non-CMFT service users may prefer to determine their own laboratory procedures pertaining to the fixation and handling of the tissue samples that send to us for examination, we ask all service users to appreciate that fixation and handling procedures affording perfectly satisfactory light microscopy fixation, may not necessarily remain suitable for ultrastructural studies.

We therefore recommend that, in order to ensure the quality and reliability of ultrastructural examination is not compromised, all service users endeavour to meet the following conditions for all tissue samples to be processed for EM:

a) Samples are placed into fixative (10% Formalin or 2.5% buffered glutaraldehyde) immediately upon removal from the patient.

NB. Buffered glutaraldehyde is preferred. Buffer should be 0.1M phosphate buffer (pH 7.4).

b) If samples are fixed initially in Formalin, they are transferred to 2.5% buffered glutaraldehyde at the earliest opportunity and fixed at 4 - 20°C for between 24 - 72 hours, after which time the sample must be transferred to 0.1M phosphate buffer (pH 7.4) for transport (see section 4.3).

c) Samples fixed in buffered glutaraldehyde are not larger than approx. 1mm3 in size.

 

(Last updated April 7th 2016)