The CellSoft computerized semen analysis system: sperm image sizes and velocity measurements, and the consequences for internal running conditions

The CellSoft computer-assisted sperm analysis system was evaluated as to the optimal setting of three components of the internal running conditions. These 'set up' parameters were the lower setting for cell size range (PIXMIN), threshold velocity (MINVEL) and maximum velocity (MAXVEL). The decision for the PIXMIN setting was founded on observed sizes of sperm with low instant velocities combined with a study of the relationship between false positive observations for sperm and the PIXMIN setting. Our recommendation for this setting is three pixels. From the observation that 97.5% of immobilized sperm exhibit instant velocities of less than 15 microns sec-1, this is our proposal for the MINVEL setting. Correspondingly, the 97.5 percentile for instantaneous velocities in a population of some 14,000 cells (30 men with type 'a' motility, WHO) was 168 microns sec-1. We recommend that MAXVEL be set at this limit. Alternative settings for both MINVEL and MAXVEL are presented as some users may wish to apply other percentiles. From the above results a setting for PIXMAX at 25 pixels could be inferred. We recommend that these settings be applied in semen laboratories analysing human native samples at room temperature.     

The influence of pentoxifylline on motility and viability of spermatozoa from normozoospermic semen samples

In order to evaluate the effects of pentoxifylline on sperm motility and longevity, a controlled in-vitro study was conducted on normozoospermic donor semen samples using the Cellsoft automated system for sperm motility analysis. After incubation and selection, pentoxifylline was found to improve the recovery of spermatozoa and to increase their velocity. In the subgroup of progressively motile spermatozoa, curvilinear velocity was also enhanced. It is concluded that pentoxifylline has an effect on the vigour, but not on the pattern, of sperm motion. Pentoxifylline did not improve the motility characteristics of senescent spermatozoa in normozoospermic sperm samples. Sperm survival, as shown by supra-vital staining, and motility longevity both decreased with time after pentoxifylline treatment.     

Improvement in quality of cryopreserved human spermatozoa by swim-up before freezing

Selecting a population of spermatozoa by the swim-up technique yields, after freezing and thawing, a population of cells that contains proportionally more spermatozoa which are morphologically normal, fewer spermatozoa with damaged tail membranes, and a greater percentage of progressively motile spermatozoa with greater velocities and amplitudes of head displacement than those obtained after freezing and thawing the same semen samples in the normal way. This pattern was found for the semen from 10 patients and five volunteers. However, the cells selected by swim-up were as susceptible to the stresses caused by freezing and thawing as unselected spermatozoa in the original semen sample, and the improvement came from the better quality of the initial sample.     

计算机辅助精液分析男性不育患者精子运动参数与精子活力的相关性

目的探讨计算机辅助精液分析精子运动参数在评价男性不育患者精子活力中的价值。方法按《WHO人类精液及精子-宫颈黏液相互作用实验检验手册》标准,采用国产WLJY-9000伟力彩色精子质量检测系统对276例男性不育患者的精液进行平均直线运动速度、平均曲线运动速度、运动的前向性、运动的直线性、运动的摆动性、平均路径速度、精子活力及分级等进行检测并分析其相关性。结果276名男性不育患者的平均精子活力为（48.93±19.10）%,分级为A级（32.11±17.25）%、B级（17.03±8.91）%、C级（10.14±5.99）%。平均直线运动速度、平均曲线运动速度、运动的前向性、运动的直线性、运动的摆动性、平均路径速度与精子活力的相关系数分别为0.60（P〈0.01）、0.59（P〈0.01）、0.51（P〈0.01）、0.55（P〈0.01）、0.52（P〈0.01）、0.67（P〈0.01）。结论计算机辅助精液分析精子运动参数平均直线运动速度、平均曲线运动速度、平均路径速度是反映精子活力的有效指标,精子运动参数对男性不育的诊断和生育能力的评估具有实用意义。     

精子微生物动静态分析技术的新进展

本文简单介绍了荧光染色精子新技术在CASA系统中的地位、意义和它的特点，以及它在操作功能上的独到之处。     

Computer image sperm selection as a novel approach to subzonal insemination in the human

Utilizing real-time computer image analysis, individual spermatozoawere selected using microaspiration. Selection criteria werebased on potential hyperactivation motility characteristics;the amplitude of lateral head displacement >7.5 µm,curvilinear velocity >70 µm/s and linearity of <30%.For this pilot study, 16 patients (eight in each group) wererecruited. Using subzonal insemination (SUZI), up to five (mean= 4.4 ± 0.3) spermatozoa selected using computer-imagesperm selection (CISS) were microinjected, or up to 15 (mean= 12.8 ± 1.3 SD) unselected spermatozoa. In the groupwhich utilized CISS, 28 out of 49 (57%) oocytes were fertilizedcompared with 13 out of 52 (25%) utilizing conventional SUZI(P < 0.04); polyspermy was 20% (n = 10) and 2% (n = 1) respectively.CISS with SUZI showed increased efficiency in achieving fertilizationand is a novel approach to studying individual sperm functionin a sperm egg bioassay where gamete ratios are close to unity.     

Cancer associated serum antigen (CASA) levels in patients with breast carcinoma and in 3 control groups without breast cancer

We have examined the predictive value of measurements of CASA (Cancer Associated Serum Antigen) levels in patients with breast carcinoma. The measurement uses monoclonal antibodies that bind to an epitope on the polymorphic epithelial mucin. CASA levels were of less value compared with the levels of other tumor markers (CA 15-3, CA 549 and CEA) in the management of patients with early breast cancer. Received: 21 October 1997 / Accepted: 20 January 1998     

Heat-Induced Hyperactivation

Purpose: The objectives of this study were (1) to determine the sperm hyperactivation and related kinematic parameters at 40°C after using four sperm wash procedures and (2) to correlate the heat-induced hyperactivation data with cases of clinical pregnancies from either artificial insemination or standard in vitro fertilization (IVF). Methods: Semen samples (n = 51) were collected by ejaculation, and semen analyses were carried out to determine the pretreatment data. Sperm kinematic measurements were performed using the Hamilton Thorn HTM-C computer-aided sperm analyzer. Hyperactivation was determined using the sort module on the HTM-C. Membrane integrity was assessed using the hypoosmotic sperm swelling procedure. Sperm morphology and acrosomal status were also determined using the Spermac stain. Each semen specimen was divided and processed through either the swim-up wash, the 1-h test-yolk buffer (TYB) wash, the 1 mg/ml pentoxifylline stimulant procedure, or the two-layer 90:47% gradient colloidal solution procedure. The washed sperm were incubated at 25 or at 40° C for 4 hr. After incubation, kinematic parameters were assessed for the posttreatment data. Semen specimens were obtained on different occasions for artificial insemination or standard IVF. Data from intracytoplasmic sperm injection cases were not included to avoid confounding factors. Live births and/or pregnancies with fetal heartbeat examined by ultrasound were considered clinical pregnancies. Results: Heat-induced hyperactive motility was significantly higher in sperm of the male partner of pregnant (n = 7) patients compared with nonpregnant (n = 44) patients (mean ± SE, 10.0 ± 3.3 versus 5.5 ± 0.8%) after TYB processing fallowed by 4 hr of incubation at 40°C. This was also observed after colloid (Percoll) processing (11.6 ± 4.6 versus 5.8 ± 0.8%). There were no differences in hyperactivation after 4 hr at 23°C between pregnant and nonpregnant cases. Parameters such as count, volume, motility, viability, and acrosomal status were not different for the groups. However, the percentage of sperm with normal morphology (WHO classification) was twice as high in the pregnant group versus the nonpregnant group. Conclusions: Heat-induced hyperactivation was associated with fertile sperm and was predictive of pregnancy obtained after artificial insemination or IVF. The association was evident only after TYB or Percoll sperm processing. The study could not confirm the finding of significant decreases in motility after heat treatment of sperm derived from infertile males. The mechanism for heat-induced hyperactivation did not involve membrane integrity or the sperm acrosome, although an involvement of heat shock proteins was postulated. Interestingly, there were no pregnancies when sperm did not exhibit heat-induced hyperactivation.     

Sperm speed is associated with sex bias of siblings in a human population

Recent studies investigating possible causes of male subfertility have largely focused on how lifestyle or environmental factors impact on the process of spermatogenesis. Markedly, fewer studies have investigated those risk factors that result in reduced sperm quality, such as poor sperm motility. The speed at which sperm swim is a major predictor of fertility and is extremely variable in human populations. It has been hypothesized that offspring sex may be adaptively manipulated to maximize the offspring''s reproductive fitness (e.g., parents with genes for good male fertility traits, such as high sperm speed, would produce primarily sons and fewer daughters because the offspring will inherit advantageous male fertility genes). Conversely, parents with poor male fertility genes would produce primarily daughters. We tested whether there was an association between how fast a man''s sperm swam and the sex bias of his siblings in a sample of men attending clinic for fertility investigations with their partner and with a wide range of semen characteristics, including sperm speed. We found that the sex bias of a man''s siblings is associated with his sperm speed; men with female-biased siblings had significantly slower sperm (judged using computer-assisted sperm analysis (CASA)) than men from male-biased sibships. This observation suggests family composition is an important factor that needs to be considered in future epidemiological and clinical studies of human fertility.     

弗司扣林对人精子运动功能的影响

目的 通过研究弗司扣林(forskolin)对体外人精子的运动功能有无影响，了解环磷酸腺苷／蛋白激酶A(cAMP／PKA)信号传导途径是否参与人精子运动功能的调节。方法 10例健康成年男性手淫获得新鲜精液，经上游优化处理后与不同浓度的forskolin一起孵育20、30、60min后，采用计算机辅助的精子分析系统(CASA)检测精子的各项运动参数，并进行对比分析。结果 forskolin在体外能显著提高人精于的活率及前向性运动百分率，而对精子的形态及直线速度(VSL)和曲线速度(VCL)无明显影响。结论 forskolin在体外能提高人精子的运动功能，此结果为证实cAMP/PKA信号传导通道参与调节人精子运动功能提供了一定的实验依据。