Comparison of manual microscopic and computer-assisted methods for analysis of sperm count and motility.
TL;DR: In this paper, a comparison of three different commercially available computer assisted semen analyzers (CASA) was conducted to determine which of the three methods, manual analysis, and two commercially available CAEs, was the most reproducible.
Abstract: This investigation was conducted to determine which of three methods, manual analysis, and two different commercially available computer-assisted semen analyzers (CASA), was the most reproducible. Semen samples from donors participating in an artificial insemination program (n = 1) and from patients being seen for andrology procedures (n = 12) were acquired at 0.5 h after ejaculation. Each specimen was loaded into one chamber of a 20-microns microcell slide (Conception Technologies, San Diego, CA, USA) and the port was sealed with petroleum jelly to prevent drying of the specimen. The specimens were assessed for sperm count (SC) and motility (MOT) first by manual analysis using an eyepiece reticle and brightfield light microscopy at 400 x total magnification, second using the Hamilton-Thorn 2030 analyzer (Hamilton-Thorn Research, Danvers, MA, USA), and third, using the Cell Trak/S system (CTS; Motion Analysis Corporation, Santa Rosa, CA, USA). Each analysis was repeated five times for each specimen on the same microcell by the same technician. The three methods were compared in terms of means and standard deviations of the SC and MOT over repeated measures-of a specimen using sign tests. The CTS system measured significantly lower sperm counts than the HTM system. MAN was intermediate and not significantly different from either. For MOT, there were no significant differences. Comparison of the standard deviations demonstrated that the three methods were not equally reproducible. For SC, the manual method was significantly less reproducible than the HTM system; the CTS system was intermediate. For MOT, the manual method was less reproducible than either CASA system, both of which were not significantly different from each other. CASA methodology in general provides a more reproducible (less variable) analysis than the manual microscopic method for assessing sperm count and motility.
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TL;DR: This review focuses on the concept of CASA, the development course of CAS a technology, the clinical application of CASa systems and the factors influencing the accuracies of results, such as frame rate, sperm counting chambers affiliated to the CASA system, algorithms and sperm concentration.
Abstract: Computer-aided sperm analysis (CASA) system has been accepted and used commonly as a routine semen analysis instrument in hospital clinical laboratories worldwide. However, technicians in clinical laboratories have little informed knowledge about the principles of CASA system and the sources of analysis errors. In this review, we focus on the concept of CASA, the development course of CASA technology, the clinical application of CASA systems and the factors influencing the accuracies of results, such as frame rate, sperm counting chambers affiliated to the CASA system, algorithms and sperm concentration. These factors and lack of internal quality control may result in huge errors of the CASA between systems and laboratories. It is therefore necessary to perform the standardisation and quality control for CASA.
77 citations
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TL;DR: Evaluating the different kinematic (velocity) parameters of frozen/thawed bull semen to determine if any of them could be correlated with their fertilising capability after insemination based on the achieved pregnancy rate concludes that VAP is the most useful semen motility characteristic which has clinical relevance in the prediction of fertility.
Abstract: Motility is one of the most important characteristics associated with the fertilising ability of spermatozoa indicating their viability and structural integrity. Therefore, the examination of motility constitutes an integral part of semen analysis. Computer-assisted semen analysis (CASA) allows an accurate and objective assessment of different sperm motion characteristics with high repeatability. The aim of this study was to evaluate the different kinematic (velocity) parameters of frozen/thawed bull semen and determine if any of them could be correlated with their fertilising capability after insemination based on the achieved pregnancy rate. Ejaculates from 10 bulls were collected and frozen. The kinematic/velocity parameters of spermatozoa were measured by CASA and compared to the pregnancy results of almost 9,000 females artificially inseminated (AI) with frozen semen of any of the 10 tested bulls. The data of the experiments are summarised mainly with a focus on the effects of individual velocities (curvilinear velocity: VCL, straight-line velocity: VSL, average path velocity: VAP) on fertility rather than on the influence of progressive motility as a whole. We conclude that VAP is the most useful semen motility characteristic which has clinical relevance in the prediction of fertility.
47 citations
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TL;DR: In this paper , the authors provided the Sperm Videos and Images Analysis (SVIA) dataset, including three different subsets, including subset-A, subset-B and subset-C, to test and evaluate different computer vision techniques in computer aided sperm analysis.
Abstract: Computer-Aided Sperm Analysis (CASA) is a widely studied topic in the diagnosis and treatment of male reproductive health. Although CASA has been evolving, there is still a lack of publicly available large-scale image datasets for CASA. To fill this gap, we provide the Sperm Videos and Images Analysis (SVIA) dataset, including three different subsets, subset-A, subset-B and subset-C, to test and evaluate different computer vision techniques in CASA. For subset-A, in order to test and evaluate the effectiveness of SVIA dataset for object detection, we use five representative object detection models and four commonly used evaluation metrics. For subset-B, in order to test and evaluate the effectiveness of SVIA dataset for image segmentation, we used eight representative methods and three standard evaluation metrics. Moreover, to test and evaluate the effectiveness of SVIA dataset for object tracking, we have employed the traditional kNN with progressive sperm (PR) as an evaluation metric and two deep learning models with three standard evaluation metrics. For subset-C, to prove the effectiveness of SVIA dataset for image denoising, nine denoising filters are used to denoise thirteen kinds of noise, and the mean structural similarity is calculated for evaluation. At the same time, to test and evaluate the effectiveness of SVIA dataset for image classification, we evaluate the results of twelve convolutional neural network models and six visual transformer models using four commonly used evaluation metrics. Through a series of experimental analyses and comparisons in this paper, it can be concluded that this proposed dataset can evaluate not only the functions of object detection, image segmentation, object tracking, image denoising, and image classification but also the robustness of object detection and image classification models. Therefore, SVIA dataset can fill the gap of the lack of large-scale public datasets in CASA and promote the development of CASA. Dataset is available at: https://github.com/Demozsj/Detection-Sperm.
32 citations
Journal Article•
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TL;DR: It was shown that low cadmium concentrations have the most negative influence on straight line velocity, which suggests the possible negative influence of cadmiam on the ability of sperm to fertilize female gametes.
Abstract: The effect of different cadmium concentrations on sperm motility in common carp was investigated. The motile activity of spermatozoa was evaluated by means of computer assisted sperm analysis (CASA) using three major parameters characterizing sperm movement - VCL, VAP and VSL. Moreover, subjective microscopic observations were performed in order to evaluate the average time of sperm movement. The following cadmium concentrations were tested: 10, 50 100, 200, 500, 1,000 and 2,000 ppm. Computer assisted analysis and microscopic observations both showed that cadmium decreases the motility of carp spermatozoa in all tested concentrations, and that lethal effects were detectable at a concentration of 500 ppm (as determined by CASA) or 1,000 ppm (when manual microscopic observations were performed). Additionally, it was shown that low cadmium concentrations have the most negative influence on straight line velocity, which suggests the possible negative influence of cadmium on the ability of sperm to fertilize female gametes.
25 citations
Journal Article•
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TL;DR: Computer-assisted sperm analysis and subjective microscopic observations showed that zinc decreased the motility of sperm, having lethal effects at concentrations of 500 ppm or 200 ppm, suggesting the possibility of its negative impact on the fertilisation rate.
Abstract: The influence of zinc on common carp sperm motility was investigated using a computer assisted sperm analysis (CASA) as well as subjective microscopic measurements of time of motility. The analysis by CASA method consisted of 3 parameters - curvilinear velocity (VCL), average path velocity (VAP) and straight line velocity (VSL), The following zinc concentrations were used: 10, 50, 100, 200, 500, 1000 and 2000 ppm. Both, CASA and subjective microscopic observations of sperm motility showed that zinc decreased the motility of sperm, having lethal effects at concentrations of 500 ppm (as analysed by computer assisted methods) or 200 ppm (as evaluated using microscopic observations of time of motility). Moreover, the computer-assisted analysis demonstrated that zinc had the strongest inhibitory influence on VSL, suggesting the possibility of its negative impact on the fertilisation rate. CASA method seems to be more reliable than classical microscopic observation of sperm activity.
25 citations
References
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TL;DR: Frozen‐thawed sperm may undergo premature break‐down of the acrosome prior to interaction with the oocyte, thus explaining the reduced fertility potential of cryopreserved semen.
Abstract: The purpose of this study was to use fluorescence microscopy to determine the viability and acrosome status of fresh and frozen-thawed human spermatozoa. Sperm cells were stained with the viability stains Hoechst 33258 (H33258) alone, or propidium iodide (PI) alone, and PI in combination with FITC-conjugated Pisum sativum agglutinin (PSA). The PSA stains the acrosome contents of permeabilized acrosome-intact sperm. Viability by fluorescence microscopy was compared to conventional eosin nigrosin staining. The overall viability using H33258 was not significantly different from that using PI. Therefore, PI was used in combination with PSA for simultaneous measurement of viability and acrosome status at the same excitation wavelength (488 nm). By combining PI and PSA, four subgroups of cells could be detected: group I, PI-neg/PSA-neg--viable, physiologic acrosome reacted (AR); group II, PI-neg/PSA-pos--viable, non-AR; group III, PI-pos/PSA-neg--nonviable, non-AR; group IV, PI-pos/PSA-neg--nonviable, degenerative AR. The postthaw sperm exhibited a significantly greater percent of sperm that were acrosome reacted (both viable and degenerative) (groups I and IV) than the fresh semen. We conclude that frozen-thawed sperm may undergo premature break-down of the acrosome prior to interaction with the oocyte, thus explaining the reduced fertility potential of cryopreserved semen.
49 citations
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TL;DR: The minimum number of spermatozoa needed for stable results, the variability of measurements and optimum methods of sampling the ejaculate were determined for one videomicrographic computer-automated semen analysis system.
Abstract: Videomicrographic computer-automated semen analysis systems allow quantitative description of sperm motility, velocity, progression, and head movement amplitude and frequency with unprecedented ease. The minimum number of spermatozoa needed for stable results, the variability of measurements and optimum methods of sampling the ejaculate were determined for one such system (Cell-Soft, CRYO Resources, New York, NY). Sampling a minimum of 225 spermatozoa yields stable measurements, and analyzing four microscope fields in triplicate provides data with the lowest coefficient of variation. The variability attributable to the instrument itself was acceptable for all measurements (6.2% to 15.4%) except mean amplitude of lateral head displacement. Limitations of these results and the potential utility of videomicrographic sperm movement analysis are discussed.
43 citations
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TL;DR: In this article, the set-up parameters of two different commercially available computerized sperm motion analyzer systems were evaluated for concentration, motility, velocity, and linearity with 46 patients and donors.
Abstract: Fresh semen specimens from 46 patients and donors were evaluated for concentration, motility, velocity, and linearity using two different commercially available computerized sperm motion analyzer systems. Although no significant differences in measurement of concentration or motility were observed, significant differences in velocity and linearity were recorded. Fourteen cryopreserved/thawed samples were assessed with the same set-up parameters as fresh specimens. When discrepancies between manual and computer counts were noted, the authors changed the set-up parameters and evaluated 33 additional specimens. Again, no differences in concentration and motility, but significant differences in velocity and linearity were observed. Interlaboratory results must be correlated and standardization of set-up parameters of various analyzers is essential.
42 citations