선택한 단어 수는 입니다.
최소 단어 이상 선택하여야 합니다.
최소 단어 이상 선택하여야 합니다.
선택한 단어 수는 30입니다.
최대 10 단어까지만 선택 가능합니다.
최대 10 단어까지만 선택 가능합니다.
제안 방법
- After centrifugation, samples were resuspended in 50 µL reaction mix containing 45 µL labeling solution and 5 µL terminal deoxynucleotidyl transferase (TdT). A positive control (incubation of fixed cells with 200 U/mL DNase I) and a negative control (replacing the DNA labeling mixture with distilled water) were used to standardize the assay. Negative controls were suspended in labeling solution without TdT.
- Our first aim was to determine the most effective glucose concentration in glycerol-free TRIS to maintain membrane integrity as well as acrosome integrity. Our second aim was to investigate the appropriate exposure time of dog sperm to glycerol-free TRIS containing glucose at 4℃ by examining DNA fragmentation.
- The dogs were caged individually, were provided water ad libitum and formulated dog food (Jindo®, Purina) twice daily, and exercised daily throughout the experiment.
- To determine the most effective exposure time of dog sperm to glycerol-free containing 0.3 M glucose for successful cryopreservation, dog sperm were cooled in extender 1 through 4℃ for 100 min, exposed to glycerol-free TRIS with 0.3 M glucose at 4℃ for 10, 30, 50, or 70 min, and then cryopreserved. After thawing at 37℃ for 25 sec, sperm motility, viability and DNA integrity were evaluated.
대상 데이터
- Spermatozoa were stained with Pisum sativum agglutinin (PSA) conjugated to fluorescein isothiocyanate (FITC). For each replicate sample, two slides were examined under a fluorescence microscope (Axio, Carl Zeiss, Germany), and approximately 200 spermatozoa were counted per slide. The percentage of spermatozoa with an intact acrosome (green fluorescence on sperm anterior acrosomal region) was calculated (Fig 2).
- , Eugene, OR, USA). For each replicate sample, two slides were prepared, and approximately 200 spermatozoa were counted per slide. The number of green or red fluorescent spermatozoa was counted under a fluorescence microscope (Axio, Carl Zeiss, Germany) fitted with a 488 nm excitation filter, and the percentage of membrane-intact spermatozoa (green fluorescence in sperm head) was calculated (Fig 1).
- Semen was collected from six dogs (two mixed breed, one poodle, two Shih-Tzu, and one Welsh corgi), ranging between 2 and 4 years of age. The dogs used in this study were treated and received care under the Guiding Principles for the Care and Use of Research Animals, as established by Chonbuk National University.
- Seven replicates were conducted for each experiment. Percentage data were subjected to arcsine transformation before analysis.
데이터처리
- All data are presented as means ± SE and were analyzed by analyses of variance (ANOVA) followed by Duncan's multiple range test using Statistical Analysis System ver. 8x software (SAS, Cary, NC, USA).
이론/모형
- Acrosome integrity was determined using the method described by Yu and Leibo (25). Spermatozoa were stained with Pisum sativum agglutinin (PSA) conjugated to fluorescein isothiocyanate (FITC).
- Sperm plasma membrane integrity was assessed using a 6-carboxyfluoresceindiacetate (6-CFDA)/propidium iodide (PI) fluorescent staining technique (17). Five hundred microliter aliquots of sperm suspension (5 × 105/mL spermatozoa) were mixed with 5 µL 6-CFDA (1 µg/mL) and 5 µL PI (0.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.