不同固定与染色条件对公牛精液抹片效果的影响研究OA

This study aimed to optimize semen smear preparation methods for sperm morphological abnormality assessment,thereby improving the accuracy and efficiency of detection.By comparing fixation times(0.5,1,2,and 3 min),staining times(1,2,3,and 5 min),semen volumes(5,8,and 10 μL),and staining reagents(gentian violet and crystal violet),the effects of these parameters on smear background intensity and sperm staining quality were assessed.The results showed that smears prepared with 5 μL semen yielded better outcomes than those prepared with 8 μL and 10 μL.At this volume,gentian violet produced superior staining effects compared with crystal violet.Fixation for 3 min combined with staining for 3 or 5 min resulted in darker backgrounds,while fixation for 0.5 min led to insufficient staining of spermatozoa.Considering both staining quality and time efficiency,fixation for 1～2 min and staining for 1～2 min were identified as the optimal parameter range.Ultimately,use 5 μL of semen,fixation for 1 min with 95%ethanol and staining with 0.1%gentian violet for 1 min was determined to be the optimal combination,producing smears with clear backgrounds and well-defined sperm morphology.This protocol meets the requirements for sperm abnormality assessment and has considerable potential for practical application.