Comparison of the Sperm Quality Analyzer IIC variables with the computer-aided sperm analysis estimates

Sperm Quality Analyzer (SQA) IIC, an upgrade version, is an inexpensive device and provides a quantitative estimation of sperm motility, whereas the use of computer-aided sperm analysis (CASA) provides high precision and provision of quantitative data on sperm kinetics. The aim of the present study was to evaluate if the SQA IIC variables correlated with the CASA estimates. Semen quality analysis of 71 fresh semen samples was performed using SQA IIC and CASA. Total sperm concentration, percentage of progressively motile sperm, percentage of normal morphology, motile sperm concentration, sperm motility index (SMI) and functional sperm count (FSC) determinations were performed using SQA IIC. Sperm concentration, sperm motility, and sperm motion variables including amplitude of lateral head displacement (ALH), beat cross frequency (BCF), curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN=VSL/VCL), and straightness (STR=VSL/VAP) were evaluated simultaneously on the same semen samples using CASA. The sperm characteristics were compared between SQA IIC and CASA. There were significant correlations of sperm concentration (r=0.634, p < 0.0001), sperm motility (r=0.697, p < 0.0001), and motile sperm concentration (r=0.757, p < 0.0001) between the two devices. Both SMI and FSC significantly correlated with eight CASA estimates, including sperm concentration, sperm motility, motile sperm concentration, ALH, VCL, VSL, VAP, and Rapid. SQA IIC is simple and easy to use. Moreover, the SQA IIC variables well correlated with the CASA estimates. As a screening test for semen quality, SQA IIC is considered as useful in the management of male infertility.     

Computer-assisted analysis of sperm morphology with the aid of lectin staining

Lectins are useful for staining the acrosome, which is a pre-requisite for the assessment of acrosome reaction in vitro. We tested wheat germ agglutinin, peanut agglutinin and pisum sativum agglutinin. The determination of the categories of normal and abnormal spermatozoa as defined by the World Health Organization, including an additional category 'acrosome-less cell', was performed with the aid of a system of computer-assisted semen analysis (CASA) in comparison with visual estimation. The acrosome reaction in vitro was induced by calcium ionophore A23187. Incubation with A23187 decreased the percentage of normal sperm heads and increased the number of acrosome-less sperm heads in comparison with the control samples. This shift was demonstrable with all three lectin staining procedures. No significant differences were observed in the comparison of sperm classes obtained by visual assessment or determination by CASA with two of the lectin staining procedures. After staining with pisum sativum agglutinin the classes of normal and of acrosome-less spermatozoa were significantly different between visual and CASA estimation. Our results indicate that estimation of the acrosomal status in vitro is possible by CASA when lectin staining is used.     

The variability of measuring sperm concentration and motility as determined by computer assisted image analysis and visual estimation

Summary. Systems of computer-assisted image processing (CAIP) offer good reproducibility and low intra-assay variability between replicate analyses of the same sample. The correlation between these values and those obtained by directly microscopic observation can be well demonstrated. The comparability between systems of different manufacturers is sufficient and satisfactory. Thus, they are well-suited for the routine work in the andrologic laboratory.
Since sperm parameters are used for the prediction of male fertility, it is important to know whether computer-assisted analysis improves the predictive power as compared to visual estimation. In our study we compared the variation of semen parameters between two semen samples of the same individual taken in an interval of at least 3 months, by using the CAIP and visual methods.
Significantly close correlations between the two values were obtained by both methods. Values of cell concentration showed the highest correlation in both methods ( r =0.75). The lowest correlation, but also statistically significant, was observed in the values of locally motile cells ( r = 0.54 for CAIP and r =0.30 for visual determination). Although apparantly higher in CAIP within method correlation coefficients did not show statistically meaningful differences between both methods.
We conclude from our study that if values of sperm concentration and motility as measured by CAIP are compared to those obtained by visual measurement, the first method is not superior to the latter in terms of retest reliability.     

Comparison between a conventional index of progressive sperm motility and movement variables analysed by CellSoft®

Two different techniques of human sperm motion analysis were compared. In 25 ejaculates, diluted to an appropriate concentration, the sperm motion index called "specific progressive motility" (SPM) was assessed by a modified conventional method. The same sperm preparations were also analysed by the CellSoft system for computer assisted sperm analysis (CASA). The SPM-value was found to correspond mainly to the variables motility and curvilinear velocity presented by CASA (multiple r = 0.92, P less than 0.001). Linearity of sperm progression, lateral head displacement, and beat cross frequency did not significantly contribute to the relationship between SPM and CASA. For comparative studies between SPM and CASA the logarithmic values of SPM are recommended.     

Application of the upgraded image superimposition system (SIAS) to the assessment of sperm kinematics

Dedicated software for semiautomatic objective motility assessment, based on image superimposition (SIAS), was upgraded in order to allow automatic reading of sperm tracks whilst keeping the advantages of visual check instead of the digital images of classic computer-aided sperm analysis (CASA) systems. The new system is very fast and accurate and allows an excellent reading of sperm motility in standard semen analysis. Moreover, using this system, an attempt was made to define numerically straight-line velocity (VSL), curvilinear velocity (VCL) and linearity (LIN) in each WHO sperm motility class. At 21 frames sec-1, ROC curves analysis determined the following: Class 1 (typical of WHO grade 'a'): VSL > or = 23 microns sec-1 and LIN > or = 0.58; class 2 (typical of WHO grade 'b'): VSL > 10 and < 23 microns sec-1 and LIN > or = 0.58; class 3: VSL > 10 microns sec-1 and LIN < 0.58 (this additional class was added to differentiate nonstraight progressive motility from classes 1 and 2); class 4 (typical of WHO grade 'c'): VSL < or = 10 microns sec-1. The numerical definition of sperm motility classes may contribute towards standardization in the objective evaluation of sperm kinematics.     

Evaluation of Hamilton-Thorn automated semen analysis system.

Sperm concentration and percentage motility values generated by the HT M-2030 system (CASA) were compared with those obtained by subjective semen analysis (SSA). Three calibrations arbitrarily designated as A, M, and D for gates and discriminators and three calibrations arbitrarily designated LI, MI, and HI for default pixel count (DPC) and default intensity (DI) were evaluated. The best correlation between CASA and SSA was observed utilizing M calibration (n = 114) with which sperm concentration was +0.3 x 10(6)/mL (r = .96) and motility was -6.3% (r = .89) compared to the values obtained by SSA. It was found that 35.9% of sperm concentration values and 34.2% of sperm motility values were within 10% of the values obtained by SSA. When sperm concentration was between 50 and 100 x 10(6)/mL the difference in motility was reduced (-3.2%) while the difference in sperm concentration was reversed (-2.6 x 10(6)/mL). LI calibration (DPC = 4, DI = 86) gave acceptable results with M calibration for sperm concentration (+2.1 x 10(6)/mL) and motility (-6.9%) compared to the values obtained by SSA. In the presence of sperm clumping, the difference between CASA and SSA was reversed for sperm concentration (+0.56 x 10(6)/mL for normal samples vs. -2.2 x 10(6)/mL for samples with clumping) and was reduced for sperm motility (-7.14% vs. -4.55%, respectively). HT M-2030 under proper calibration can be used as a rapid, objective, and reliable alternative to conventional semen analysis in routine and for research purposes.     

Usefulness of sperm quality analyzer-V (SQA-V) for the assessment of sperm quality in infertile men

The aim of this study was to evaluate the correlation of new of Sperm Quality Analyzer (SQA-V) with the computer-assisted sperm analysis (CASA) and manual semen analysis estimates. One hundred five fresh semen samples were analyzed using SQA-V and CASA and manual semen analysis. Significant correlations of sperm concentration (p < 0.0001), sperm motility (p < 0.0001), and normal morphology (p < 0.0001) were observed between SQA-V variables and manual semen analysis estimates. There also were significant correlations of sperm concentration (p < 0.0001), sperm motility (p < 0.0001), and sperm velocity (p = 0.0235) between SQA-V variables and CASA estimates. Meanwhile, it did not correlate with amplitude of lateral head displacement, beat cross frequency, lineality assessed by CASA. The value of the sperm concentration and the sperm motility measured by SQA-V showed high correlations with the value of those measured by CASA and manual semen analysis. In addition, velocity and sperm morphology may also be evaluated to some extent using SQA-V.     

Semen analysis and external quality control schemes for semen analysis need global standardization

One semen analysis laboratory [the Institute of Reproductive Medicine (IRM), Münster, Germany] was enrolled in three external quality assurance programmes in Europe (United Kingdom External Quality Assurance Scheme, European Academy of Andrology, European Society of Human Reproduction) that control for the assessment of sperm concentration, sperm motility and sperm morphology. Agreement between the participating laboratory and the sperm concentrations designated by all three programmes was good. Disagreement between two quality control (QC) programmes providing motility assessment was particularly noticeable in the distinction between motility grades a and b. For the identification of normal sperm morphology, marked differences between the standards set by different QC programmes were apparent. One programme was stricter in its categorization of normal forms, such that an overestimation of normal forms was diagnosed at IRM, whereas agreement with the other programmes was achieved. Variation of results from technicians in the IRM internal QC programme was <13%. The discrepancies between external quality control (EQC) centres demonstrated here are challenging problems to be overcome partly by the andrology laboratories and partly by the providers of EQC services. The introduction of objective, computer-aided sperm assessment in providing designated values may help to rectify this situation. Until this is achieved, EQC programmes should develop an internal programme to monitor their materials and methods for assessment.     

Computer‐aided sperm analysis,the new key player in routine sperm assessment

For sperm analysis, important inter‐laboratory variations have been observed in manual analyses. In this study, a computer‐aided sperm analysis (CASA) system was assessed versus manual technique, and specific software modifications were operated to fit the David's classification already used in the laboratory. Four parameters were studied (concentration, motility, vitality and morphology), and at least 30 semen samples from 30 different patients have been tested. Manual and automated analyses were compared using a least‐squares regression line analysis, Student's t test, Bland–Altman plots and Passing–Bablok regressions. Repeatability was also assessed, and coefficients of variation (CV) were calculated. Both manual and automated methods gave similar results for sperm concentration (n = 150), motility (n = 30), vitality (n = 90) and morphology (n = 90). Repeatability always showed a decrease in the CV with automated analysis; for example in normal range of sperm values, CV for manual and CASA analyses were, respectively, 9.0% versus 4.4% for sperm concentration, 5.2% versus 4.1% for motility, 7.3% versus 4.2% for vitality and 11.4% versus 4.1% for morphology. All parameters were comparable between automated and manual analysis, and repeatability measures confirm the more reliable values of the SCA compared to those of manual analysis.     

Variations in quality of frozen-thawed semen from Swedish Red and White AI sires at 1 and 4 years of age

The predictability of semen quality of mature sires from measurements at an early age is not well established. The aim of the present study was to determine age-dependent changes in the quality of bull semen for artificial insemination (AI). Semen was collected and frozen from each of six Swedish Red and White (SRB) dairy AI bulls when they were 1 and 4 years old. Three batches were randomly selected from each bull and age group. From each batch, semen was analysed immediately after thawing [post-thaw (PT), control] as well as after washing/resuspension (W) and after a swim-up procedure (SU). The analyses comprised subjective and computerized (computer-aided sperm analysis, CASA) measurements of motility as well as sperm concentration, morphology and membrane integrity. When semen was analysed, PT, overall sperm motility (CASA), concentration of motile spermatozoa and membrane integrity improved when sires were older. After SU, there was a similar improvement in membrane integrity and concentration of motile spermatozoa, but linear motility decreased. No significant differences between ages were recorded after W-treatment. The above findings indicate that SU is not only superior to W-treatment in differentiating semen quality among bulls but also reveals age-dependent changes. Improved motility and membrane integrity suggest increased viability of spermatozoa at 4 years of age in the SRB sires examined here.     

The validity and reliability of computer-aided semen analyzers in performing semen analysis: a systematic review

BackgroundComputer-aided sperm analyzers (CASA) are currently used worldwide for semen analysis. However, there are doubts about their reliability to fully substitute the human operator. Therefore, this study aimed to systematically review the current literature comparing results from semen evaluation by both CASA-based and manual approaches.MethodsA systematic screening of the literature was performed based on the PRISMA guidelines and by searching on PubMed, Scopus, and Embase databases.ResultsA total of 14 studies were included. Our results showed a high degree of correlation for sperm concentration and motility when analysis was performed either manually or by using a CASA system. However, CASA results showed increased variability in low (<15 million/mL) and high (>60 million/mL) concentration specimens, while sperm motility assessment was inaccurate in samples with higher concentration or in the presence of non-sperm cells and debris. Morphology results showed the highest level of difference, due to the high amount of heterogeneity seen between the shapes of the spermatozoa either in one sample or across multiple samples from the same subject.ConclusionsOverall, our study suggests CASA systems as a valid alternative for the evaluation of semen parameters in clinical practice, especially for sperm concentration and motility. However, further technological improvements are required before these devices can one day completely replace the human operator. Artificial intelligence-based CASA devices promise to offer higher efficiency of the analysis and improve the reliability of results.     

Standardization and comparability of CASA instruments.

Thirty human semen specimens were analyzed using a standard manual method, then videotaped and reanalyzed using two different computer-aided sperm analysis (CASA) instruments (the HTM system, Hamilton-Thorn Research, Danvers, MA, and the CTS system, Motion Analysis Corp., Santa Rosa, CA). Videotaped specimens were analyzed by CASA for 5 frames for sperm concentration (CON) and percent motility (MOT), and for 15 frames for kinematic variables (straight-line velocity, VSL; curvilinear velocity, VCL; linearity, LIN; and amplitude of lateral head displacement, ALH). Machine parameter settings for the two instruments were matched as closely as possible. CASA values were compared with each other for all measures and with manual results for sperm count and the percentage of motile sperm. Results show: 1) HTM and CTS average values for CON are not different from manual measures for the 5- or 15-frame analysis, but slight differences are seen between CTS and HTM; 2) average values for MOT for the 5-frame analysis are higher than the 15-frame analysis for both instruments, but the average manual measurement for percent motility is much higher than any CASA value; 3) average VSL and LIN are slightly higher for HTM than CTS, but pair-wise comparison shows a high degree of concordance between the instruments; and 4) the mean values for VCL and ALH are equal for the two instruments, and there is a close concordance for the pair-wise comparison for VCL; however, pair-wise comparison of ALH reveals significant differences between the instruments. Overall, the differences seen between these instruments are slight, and are probably not biologically or clinically significant.     

精液体外处理对精子核DNA链完整性影响的研究

目的 研究3种不同精子优选技术对精子运动参数及DNA链完整性的影响。方法 通过计算机辅助精液分析及彗星试验从精子运动参数及DNA链完整性两个方面评价精液体外处理对精子的影响。结果 上游法与Percoll密度梯度离心法相比,除前者精子回收率(15.499.39)%明显低于后者(42.807.17)%外,P<0.05,其他差别无显著性,PureSperm密度梯度离心法与Percoll密度梯度离心法比较,各运动参数差别无显著性(P>0.05)。精液经Percoll法及PureSperm法密度梯度离心和上游法处理后,总彗星细胞率较处理前降低(P<0.05)。结论 上游法、密度梯度离心法和Puresperm 均可以不同程度优化精液质量;对精子DNA链的损伤程度不同。     

用传统方法与计算机辅助精液分析软件CRISMAS分析166名丹麦青年男性精液样本的结果比较

本研究以166名丹麦青年男性精液为样本,通过评估精于浓度和活力来比较传统精液分析与计算机辅助精液分析方法（CASA）（哥本哈根Rigshospitalet图像屋精子运动分析系统,CRISMAS软件4．6版本）。CRISMAS软件测定精子的浓度把精子活力分为三类。传统分析方法将精子活力分为四种状态。为了便于二者的比较,本文将传统的四种状态根据精子速度等级重新分为三个状态：rapidly progressive（A）,slowly progressive（B）和non-progressive（C＋D）。两种方法所研究的参数之间都有显著差异（P〈0．001）。与传统方法相比,CRISMAS高估了精子浓度以及快速运动精子的比例,因而低估了慢速运动和非运动精子。为分析研究结果是否会随精液分析时间而起伏变动,将精液分析结果按分析同期分为四个层次。结果表明CRISMAS对活力的分析结果比传统分析方法稳定,但两种方法都未表现出任何趋势。显然,无法比较CRISMAS CASA和传统分析方法在精子浓度和精子活力方面的分析结果。在临床上使用该软件时以及用其研究这些精子特性时需要说明这一点。     

Visual versus cinemicrographic evaluation of human sperm motility and morphology

Ratings of human sperm motility by visual estimation through the microscope remain important measures of semen quality and of male fertility. More objective methods, including cinemicrography, time lapse photography, and videomicrography, are advocated. Subjective (visual) and objective (cinemicrographic) ratings of motility were compared. Sixty workers in 30 laboratories rated motilities of 40 specimens on motion picture film, and motilities were also measured by cinephotomicrographic methods. The morphology of each of the motile and immotile sperm was rated. In 34 of 40 specimens visual ratings were higher (range = +2 to +31%) than actual percentage motility. Specimens with both high sperm concentration and forward progression received the highest overestimations by visual rating. This was especially apparent in specimens with the highest motility. There was a statistically significant positive relationship between sperm motility and morphology rated on a one-by-one basis, but the relationship was too small to influence the visual rating of human sperm motility.     

The influence of sperm density on the motility characteristics of washed human spermatozoa

To study the effects of sperm density on the results of computer-assisted semen analysis (CASA), 10 washed semen samples were diluted and measured with the CellTrak/S CASA system in a concentration range of 10–180×10⁶ spermatozoa/ ml. All sperm motility parameters were influenced to some extent by sperm density. The motility percentage was influenced significantly in 5 samples ( P< 0.005), the straight line velocity in all samples (P<0.0005 in 7 samples), the curvilinear velocity in 3 samples ( P< 0.005), the linearity in 9 samples (P<0.0005 in 6 samples) and the lateral head displacement in 9 samples (P< 0.005 in 6 samples). In general, the CellTrak/S data are influenced significantly if sperm density exceeds 50 × 10⁶ spermatozoa/ml.
The influence of sperm density on the motility parameters can be explained both by the accuracy of the CASA system and by actual changes in the motility of the spermatozoa. In the light of other published studies, it is concluded that sperm motility measurements with CASA systems should be assessed using 25–50 × 10⁶ spermatozoa/ml, especially in studies concerning lateral head displacement and the linearity, as in sperm hyperactivation studies.     

CASA as an aid to selecting sperm suspensions for artificial insemination in Callithrix jacchus

The ability to use sperm motility parameters, obtained by computer-assisted sperm motility analysis (CASA), as an aid to selecting sperm samples for artificial insemination (AI) would have considerable benefits for commercial organizations and for the captive breeding of endangered species. In this study the Hobson sperm tracker (HST) was validated for use with spermatozoa from Callithrix jacchus, the common marmoset, by comparing values for straight line velocity by CASA with those obtained by direct measurement of sperm tracks. Using the settings established during validation, ejaculated and epididymal spermatozoa were analysed with the HST. The range of values for velocity parameters were used to establish expected motility profiles for the two types of spermatozoa as follows: for epididymal spermatozoa (concentration 2.2 - 85.8 x 10(6) spermatozoa/mL) VCL 109.8-155.9, VSL 75.2-141.5, VAP 85.8-142.1, MAD 13.7-40.7, ALH 3.8-7.8, BCF 1.4-4.2, LIN 40.5-91.1% and STR 70.1-97.1%; for ejaculated spermatozoa (concentration 3.2-82.0 x 10(6) spermatozoa/mL) VCL 89.6-136.7, VSL 69.6-110.3, VAP 74.5-121.9, MAD 19.2-29.3, ALH 3.0-9.9, BCF 2.8-5.5., LIN 65.4-85.3% and STR 93.8-97.7%. Epididymal spermatozoa from males which were not sexually active had significantly lower values for VCL, VSL and VAP, while values for MAD were significantly higher than for spermatozoa from sexually active males (p < 0.031). Sperm concentration affected motility parameters significantly. Although motility parameters differed according to the batch of medium used, the differences were not statistically significant. Epididymal sperm samples had significantly higher VCL, VSL and VAP but lower BCF and LIN than ejaculated sperm samples of the same concentration diluted in the same batch of medium, while MAD, ALH and STR were not different. Urine contamination significantly reduced VCL, VSL and VAP (p < 0.008, < 0.016 and < 0.008, respectively, sample size = 7) whereas MAD, ALH, BCF, LIN and STR were not affected. Therefore CASA could be useful in screening ejaculates for use in Al to eliminate samples with unusual motility patterns.     

手动或计算机辅助方法评估经间歇性阴囊隔离的公羊的精子活力(英文)

目的:研究用计算机辅助的精液分析(CASA)技术辅助测量得到的精子运动特征是否比手动检测得到的活动的、快速的或前进的细胞百分比能更敏感地指示精液质量下降。方法:6只公羊每天进行16 h 间歇性阴囊隔离,持续21天,对其中2只在次年进行每天12 h 的间歇性阴囊隔离,持续28天。收集其精液并立即进行 CASA 评估,然后冷冻或在30℃或5℃的条件下储藏,用于再次评估。结果:间歇性阴囊隔离导致运动的、快速的和前进的精子百分比下降,与冷冻—解冻或储存在30℃或5℃下的情况相同。只有当运动精子的百分比下降时,通过 CASA 测定的精子能动性(精子侧摆幅度、平均路径速度、平均前进速度和轨迹速度)才降低。冷冻—解冻或液态储存导致从阴囊隔离公羊的精液中有运动能力和快速的精子百分比较之对照组下降幅度更大,但是仅在有运动能力的精子百分比下降时才影响精子的运动特征。结论:间歇性阴囊隔离不仅影响新鲜收集的精子运动,而且影响其抵抗外界储存压力的能力。而通过 CASA技术获得的其他关于运动特征的数据仅是精液质量下降时运动细胞百分比的一个敏感指示。     

Comparison of semen analysis between the two Hamilton-Thorn semen analysers

This study was designed to assess the reproducibility of the computer-assisted semen analysis (CASA) system. Semen specimens from 41 suspected subfertile patients were analysed on two identical Hamilton-Thorn Motility analysers (HTM, model 2030, version 7). Specimens were analysed after liquefaction at 37 degrees C for 30 min on two HTMs adjusted to the same gate settings and by using the same semen droplet loaded on a Makler counting chamber prewarmed at 37 degrees C. Significant differences were seen in the total sperm count (P < 0.0004), motile sperm count (P < 0.004), amplitude of lateral head displacement (P < 0.0001), linearity (P < 0.01), and beat cross frequency (P < 0.0001) between the two HTMs. No significant differences between the two machines was seen in the case of other sperm motion parameters: percentage motility, average path velocity, straightline velocity, curvilinear velocity, straightness, and progressive motility. Our results indicate that the reproducibility of several semen parameters analysed by HTM is poor, and steps to remedy this problem are necessary.