The main kinetic parameters measured are the straight-line velocity (VSL), curvilinear velocity (VCL) and average path velocity (VAP).

Individual spermatozoa trajectories are based on spermatozoa head centroid position in consecutive frames of video recordings, which is correlated with the flagellar movement patterns 3, 9, 10, 11. However, a limiting factor of motility parameter is the frame rate of the video camera. This technology includes the development of software associated with a phase contrast microscope that allows the assessment of sperm motility. However, there are many innovative, rapid and quantitative techniques that can provide a reliable sperm quality analysis 2, 4.Д¬omputer-assisted sperm analysis was developed to offer accurate data about sperm quality 8. Direct observation of sperm motility can produce inaccurate and subjective data as it requires time and experience, which leads to a lack of consistency and incompatibility of results 6, 7. In this sense, the correct evaluation of individual ejaculate/spermatozoa features, such as motility, is fundamental to obtain discriminatory information regarding individual male fertility potential. However, the sperm variability between males can lead to sperm competition and, consequently, not all the males are equally contributing to the gene pool 5. Mixing sperm from multiple males is a common practice in the production of many aquatic commercial species 4. The convenient evaluation of gamete quality is the best tool for defining the fertility potential of a specimen. This is the major factor that contributes to successful fertilization, allowing the development of viable offspring 3, 4. The efficacy of reproduction depends on the quality of both gametes (eggs and sperm) 1, 2. This protocol describes the analysis of fish sperm motility using software for sperm analysis and new cooling devices to optimize the results. Therefore, cooling devices are necessary to refrigerate samples at constant temperature over the time of analysis, including on the optical microscope. However, for some fish species, it is advantageous to use a lower temperature to prolong the duration of motility, since the sperm remain active for less than 2 min. For mammalian sperm, the heating plate is used to maintain optimal temperatures of spermatozoa. Thus, it is necessary to modify both software and hardware components to make motility analysis more efficient for fish sperm analysis. Fish have specific features that can affect sperm assessment such as a short motility time after activation and, in some cases, adaptation to lower temperatures. Initially, this computer technology was designed for mammalian species, although it can also be used for fish sperm analysis. Computerized systems for sperm analysis were developed to measure several parameters and one of the most commonly measured is the sperm motility. Morphology analysis perform parametric study of sperm.For gamete quality evaluation, there are innovative, rapid, and quantitative techniques that can provide useful data for aquaculture. Motility measures the count and percentage of fast moving, slow moving and dead(immotile) sperm.ДЄlso measures the overall count and percentage of motile and immotile sperms.

Motility analysis is determined by computing the sperm cells movement and its path in the frame.

Motility module performs motility (motion) analysis of the sperms from the input video of any time period. Measurement result can be printed with patient's details, graphs, measurement images. Reduce the operational cost by using reusable counting chamber. Most of analysis are done with single button click. Suitable for hospitals, fertility clinics and Labs to evaluate cases of male infertility and other practices. 'Sperm Pro' follows WHO Laboratory manual. This software analyse accurate, repetitive and automatic assessment of following sperm Parameters: Motility, Concentration, Morphology, DNA Fragmentation & Vitality. Computer Aided Sperm Analysis (CASA) software suitable for human sperm.

Casa software sperm