Epididymal sperm analysis, including motility, count and abnormal morphology rates, was carried out as described previously [13 (link)]. The left epididymis of each rat was removed. Epididymal plasma was obtained by cutting the caudal part of the left epididymis in a petri dish. The seminal plasma was kept for two minutes at room temperature to be liquefied. Semen sample was taken into GMOPS plus (Vitrolife, USA) solution. After centrifugation using the routine gradient method, the pellet was centrifuged again by dilution with the sperm washing medium (Vitrolife, USA). The sperm count and motility in the Makler chamber (Sefi Medical Instruments, Haifa, Israel) were evaluated using the Olympus BX43 microscope (Tokyo, Japan). The semen fluid was spread on the lam and left to dry. It was subsequently dyed by Diff Quick (NBT Lab) staining method for morphological evaluation. Under x100 magnification using the immersion oil, the total area of 100 spermatozoa was counted and percentage values regarding the disorders in the morphology and structure of head or tail were given.
Sperm washing medium
Manufactured by Vitrolife
Sourced in United States, Sweden
Sperm washing medium is a laboratory solution used to prepare sperm samples for assisted reproductive techniques. It is designed to separate and concentrate sperm cells from the seminal plasma, while maintaining their viability and motility.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using sperm washing medium
1
Comprehensive Epididymal Sperm Evaluation
2
Semen Analysis and Sperm Selection Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh ejaculated semen was obtained by masturbation and collected on the oocyte retrieval day in sterile containers and kept for half an hour at 37∘C for liquefaction followed by concentration, total motility, viability, and morphology analyses under the light microscope. The semen analysis was based on the standard of the fifth edition of the WHO guidelines. A normal semen sample should be equipped with at least a concentration of 15 × 106/ml, a total motility of 40%, and a normal morphology rate of 4% (Xin et al., 2020 (link)). Standard density-gradient centrifugation was performed for sperm selection as previously described (Huang et al., 2015 (link)). Briefly, 1.5 mL of 45% gradient media (Vitrolife, Gothenburg, Sweden) was added on the top of 1.5 mL of 90% gradient media. Then, up to 3 mL of the semen was layered on the gradient media and centrifuged at 200 g at room temperature for 20 min. The sperm pellet was isolated and washed with 3 mL of Sperm Washing Medium (Vitrolife, Gothenburg, Sweden) at 300 g for 6 min. Then, the sperm pellet was resuspended in 0.5 mL of Sperm Washing Medium and left at room temperature to allow for a swim-up for 30–60 min. The top 300 μL was collected for semen parameters analysis.
3
Sperm Analysis and IVF Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A combination of manual Papanicolaou sperm staining and a computer-assisted sperm analysis (CASA) system (BEION S3-3, V4.20; BEION, Shanghai, China) was applied in semen analysis. Quality control of CASA was conducted every day to avoid any possible biases, and a standard semen analysis according to the World Health Organization (WHO) Semen Analysis Manual 2010 [7 (link)] was performed before CASA. The lower reference limits of semen parameters were based on the WHO criteria (fifth edition), namely semen volume (1.5 mL), sperm concentration (15×106/mL), total sperm count (39×106 per ejaculate), total motility (40%), progressive motility (32%), and normal forms (4%) [7 (link)].
Semen preparation for IVF was selected using standard density-gradient centrifugation. Up to 3 mL semen was added to 3 mL 45%/90% gradient media (1:1; Vitrolife, Gothenburg, Sweden) and centrifuged at 200 g for 20 minutes. After washing, the pellet was resuspended in 0.5 mL sperm washing medium (Vitrolife) to allow for a 30 to 60 minutes swim-up. The top 0.3 mL was collected for optimized analysis and insemination. Normally, IVF was performed when the optimized sperm had a concentration above 5×106/mL; otherwise, the sperm were subjected to ICSI.
Semen preparation for IVF was selected using standard density-gradient centrifugation. Up to 3 mL semen was added to 3 mL 45%/90% gradient media (1:1; Vitrolife, Gothenburg, Sweden) and centrifuged at 200 g for 20 minutes. After washing, the pellet was resuspended in 0.5 mL sperm washing medium (Vitrolife) to allow for a 30 to 60 minutes swim-up. The top 0.3 mL was collected for optimized analysis and insemination. Normally, IVF was performed when the optimized sperm had a concentration above 5×106/mL; otherwise, the sperm were subjected to ICSI.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!