The use of Hanks'--pipes buffers in the preparation of human, normal leucocytes for TEM observation.

It is suggested that the use of Hanks' + pipes + sucrose buffers, in combination with glutaraldehyde and osmium tetroxide fixatives, represent an excellent mode of preparation of fresh and cultured peripheral blood leucocytes, not only for transmission electron microscopy observation of these cells but also for scanning electron microscopy.

Tannic acid- and thiocarbohydrazide-mediated osmium tetroxide binding in the preparation of human leucocytes for SEM observation.

Normal human leucocytes, successively treated with glutaraldehyde-tannic acid-osmium tetroxide-thiocarbohydrazide-osmium tetroxide-thiocarbohydrazide-osmium tetroxide, were prepared for scanning electron microscopy observation. These cells produced well-contrasted, non-charging scanning images compatible with metal-evaporated material. Further, the mononuclear and polymorphonuclear cells resisted shrinkage during dehydration and critical point drying, thus allowing much improved images at high magnification than those covered with evaporated metal.

The simultaneous use of Hanks' and Hepes buffers in the preparation of human leucocytes for SEM observation.

Leucocytes were isolated from the peripheral blood of normal individuals and prepared for SEM observation. The addition of the buffer Hanks and Hepes to the glutaraldehyde fixative was found to maintain the initial ambient pH throughout cell manipulation, and to allow satisfactory preservation of surface architecture.

SEM observation of cultured, mitogen-stimulated leukocytes. Cells obtained from patients with acute myeloblastic and chronic lymphocytic leukemias.

SEM observation of acute myeloblastic leukemia cells, incubated for 20 h with the mitogens pokeweed and phytohaemagglutinin, showed these to have elongated structures that were either smooth or partially covered by thumblike figures. By contrast, the chronic lymphocytic leukemia cells possessed more compact shapes and some were covered with blebs of varying sizes.

The use of various buffers in the preparation of human lymphocytes for SEM observation.

A study is presented of various buffers utilized in the preparation of human lymphocytes for scanning electron microscopy. Of nineteen different buffers tested, Hanks' balanced salt solution +0.04 mol sucrose appeared most adequate for satisfactory preservation of lymphocyte surface architecture.