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Scanning electron microscopy of peripheral blood leukocytes of the chickens
Abstract:
Polymorphonuclear leukocytes, e.g., neutrophilic granulocytes, were enriched from heparinized blood by a Ficoll-step-gradient centrifugation procedure. Scanning electron microscopy (SEM) revealed a surface morphology of narrow ridge-like profiles and small ruffles with occasional microprocesses. Mononuclear leukocytes were isolated by centrifugation over a Ficoll-Metrizoat gradient. The lymphocytes showed varying numbers of microvilli of different length, size and shape. B lymphocytes, characterized by their capability of "sheep red blood cell (SRBC)-rosette formation", displayed a similar surface morphology. Completely smooth lymphocytes, described in the literature as T lymphocytes, could not be detected, although many lymphocytes with few microprocesses were observed. Thus, SEM is not a useful tool for distinguishing between B and T lymphocytes in the peripheral blood of chickens. Monocytes were characterized by prominent membrane-like ruffles, but in some cases they closely resembled granulocytes. An influence of the various separation media on the surface morphology of the isolated cells could not be detected when compared with cells isolated by the buffy-coat method.
Insights
Scanning electron microscopy (SEM) revealed distinct surface morphologies for chicken leukocytes. However, SEM cannot reliably differentiate between B and T lymphocytes in chicken peripheral blood.
Area of Science:
- Immunology
- Cell Biology
- Veterinary Science
Background:
- Accurate identification of leukocyte subtypes is crucial in avian immunology.
- Scanning electron microscopy (SEM) offers high-resolution surface imaging of cells.
- Previous studies have explored SEM for leukocyte characterization in various species.
Purpose of the Study:
- To characterize the surface morphology of chicken leukocytes using SEM.
- To evaluate the utility of SEM in distinguishing between B and T lymphocytes in chickens.
- To assess the impact of isolation methods on cell surface morphology.
Main Methods:
- Leukocytes were isolated from chicken blood using Ficoll-based density gradient centrifugation.
- Polymorphonuclear leukocytes (neutrophilic granulocytes) and mononuclear leukocytes were separated.
- Cell surface morphology was examined using scanning electron microscopy (SEM).
Main Results:
- Neutrophilic granulocytes exhibited ridge-like profiles and ruffles.
- Lymphocytes displayed variable microvilli; B lymphocytes (via SRBC-rosette formation) showed similar morphology to other lymphocytes.
- T lymphocytes with smooth surfaces were not definitively identified; monocytes showed ruffles but could resemble granulocytes.
- Isolation media did not significantly alter cell surface morphology compared to the buffy-coat method.
Conclusions:
- SEM reveals distinct surface features of chicken granulocytes and monocytes.
- SEM is not a reliable method for differentiating between chicken B and T lymphocytes.
- The isolation techniques employed did not introduce significant artifacts in SEM-based surface morphology analysis.
