Protocol for obtaining leukocyte- and thrombocyte-rich fibrin (L-TRF) membrane in domestic quail (Coturnix coturnix japonica)

Autologous hemoconcentrates, such as leukocyte- and platelet-rich fibrin, have been widely used in regenerative medicine; however, standardized protocols for avian species of experimental and clinical relevance remain scarce. This study aimed to standardize the preparation of leukocyte- and thrombocyte-rich fibrin (L-TRF) membranes in domestic quail (Coturnix coturnix japonica) and to evaluate their clinical and laboratory characteristics. Fourteen adult quail underwent right jugular vein venipuncture, with whole blood collected into additive-free glass tubes and centrifuged at 400 g at approximately 24 °C for 5, 10, 15, and 20 min. Membranes were obtained after partial serum absorption and characterized by light microscopy and transmission electron microscopy. The mean blood volume collected was 0.89 ± 0.37 mL (~ 0.7% of body weight). Consistent viscoelastic membranes were formed between 10 and 20 min of centrifugation. The clots exhibited a layered organization composed of fibrin, a leukocyte–thrombocyte layer, and an erythrocyte-rich fraction. Microscopic analysis demonstrated a three-dimensional fibrin network containing embedded thrombocytes and leukocytes, predominantly heterophils and lymphocytes. In conclusion, centrifugation of domestic quail whole blood at 400 g for 20 min in plain glass tubes, followed by standardized serum absorption, consistently yields cohesive and manageable L-TRF membranes. This reproducible, low-cost protocol contributes to the standardization of hemoconcentrate preparation in avian species and supports future applications in avian regenerative medicine.