Glycogen in Basophilic Leucocytes in Human Blood Smears

Abstract

In a recent study of the occurrence of the periodic acid-Schiff reaction in various normal cells of the blood and connective tissue by Wislocki, Rheingold and Dempsey, 1 small clear red dots were observed in the pale pink cytoplasm of the basophilic leucocytes. These investigators were not able to identify the basophiles in control blood smears digested by saliva. They therefore concluded that the red stained material in the undigested smears may have been glycogen but that the “apparent finding needs further verification.” Similarly, Gibb, and Stowell 2 decided that “Because myeloid cells which were free from glycogen were not observed in normal peripheral blood and bone marrow films, basophils, although not specifically identified, may also contain glycogen.”

To make certain that basophils could be identified in smears digested by saliva before treatment with periodic acid and leucofuchsin, dried smears of human blood were stained in Wright's blood stain in absolute methyl alcohol for 3 minutes. This step was not followed by the usual one of the diluted stain. For the most part, the smears were very quickly rinsed in 95% ethyl alcohol although at the beginning of the study a few were dipped in running water. Basophils were located and each cell was ringed by means of an object marker. A map was made giving its position in the circle. The smears were then fixed in absolute ethyl alcohol for 5 minutes, rinsed in 95% and 70% alcohol and stained by the periodic-leucofuchsin technic of McManus 3 with 1/2% periodic acid, or, as modified by Wislocki, Rheingold and Dempsey, 1 with 1% periodic acid. The smears to be digested were covered with saliva for 45 to 60 minutes after fixation in absolute alcohol and rinsed in running water.