Cytochemical properties of chicken blood cells resembling both thrombocytes and lymphocytes

Generally accepted criteria were used to identify typical nucleated thrombocytes and typical small lymphocytes in chicken-blood smears subjected to modified-Wright staining. Other cells, here referred to as "intermediate cells," were difficult to classify because in some aspects they resembled thrombocytes while they also had features typical of small lymphocytes. The "intermediate cells" had small, round or oval nuclei with coarsely condensed chromatin, characteristic of both thrombocytes and small lymphocytes. In addition, "intermediate cells" had moderately abundant cytoplasmic volumes, typical of thrombocytes but blue cytoplasm lacking both granules and vacuoles, which is characteristic of small lymphocytes. It made little difference to the thrombocyte count whether these cells were classified as thrombocytes or small lymphocytes; however, this decision made a substantial difference to the lymphocyte count in some chicken-blood smears. Most "intermediate cells" (351 of 410 cells examined) were nonfluorescent after treatment with formaldehyde gas. Furthermore, most "intermediate cells" failed to acquire characteristic pigments when subjected to either Grimelius staining (179 of 204 cells examined) or periodic acid-Schiff staining (173 of 206 cells examined). Typical small lymphocytes reacted in the same way, failing to fluoresce after gaseous formaldehyde treatment (65 of 65 cells examined) and failing to react during Grimelius staining (41 of 44 cells examined) or periodic acid-Schiff staining (21 of 21 cells examined). In contrast, almost all typical thrombocytes became fluorescent in response to gaseous formaldehyde (709 of 718 cells examined) and gave positive reactions when subjected to Grimelius staining (381 of 382 cells examined) or periodic acid-Schiff staining (322 of 326 cells examined). These findings suggested that "intermediate cells" should be classified as lymphocytes in differential cell counts.