Anderson (1965) has described a simple method for preparing relatively pure fields of leukocytes from peripheral blood for electron microscopy using an angular spatula to detach and remove the buffy coat from the wall of a standard conical centrifuge tube. the buffy coat is, however, fragile, especially after gentle centrifugation (1500 × g for 7-9 min) and brief (5-10 min) fixation. the spatula often breaks the buffy coat and mixes it with the red cells below. the conical centrifuge tube is also disadvantageous. Its large cross-section and capacity require relatively large amounts of blood and it gives a buffy coat which, for normal blood, is too thin to permit adequate representation in individual transverse sections of those leukocytes which occur in low percentages. the use of special tubes to circumvent these problems (Wright and Douglas 1902, Endres 1927, Bessis 1940, Butler and Cushmann 1941, Chevillard and Hamon 1943, Ottesen 1954, Bessis 1956) has been very useful for cell separation but the design of the tubes proposed is not suitable for routine electron microscopy.
ASJC Scopus subject areas
- Medical Laboratory Technology