Microfabrication of low-cost customisable counting chambers for standardised estimation of sperm concentration.

Evaluation of sperm concentration is essential for research and procedures involving AI, cryopreservation and sperm quality assessment. Microfabrication technologies have shown tremendous potential for rapid prototyping and fabrication of devices to assist reproduction and fertility research, but such utility has not yet been made available for most reproduction laboratories. The aim of this study was to evaluate the feasibility of using microfabrication techniques to produce counting chambers for estimation of sperm concentration. Zebrafish (Danio rerio) spermatozoa were used as a model for evaluation of functionality of the chambers. These microfabricated enumeration grid chambers (MEGC) were composed of a polydimethylsiloxane (PDMS) coverslip with grid patterns (100 µm×100 µm) and a PDMS base platform to create a known volume with a 10-µm height to restrict the cells to a single layer. The results of cell counts estimated by two of three prototype MEGC devices tested were not significantly different from the control device, a commercially available Makler chamber. The material cost for a MEGC was less than US$0.10 compared with product costs of approximately US$100 for a standard haemocytometer and US$700 for a Makler counting chamber. This study demonstrates the feasibility of microfabrication in creating low-cost counting chambers to enhance standardisation and strengthen interdisciplinary collaborations.

[Evaluation of practices and costs of vasectomy. French monocentric experience]. / Évaluation des pratiques et des coûts de la vasectomie. Expérience monocentrique française.

INTRODUCTION: Since the law of 4 July 2001, vasectomy has been recognized as a method of male contraception. We report the experience of vasectomy practice in a hospital-university center. METHODS: A monocentric retrospective cohort study of 45 patients who benefited from a contraceptive vasectomy between July 2001 and May 2016. For each patient were studied: modalities of implementation, compliance with the recommendations of the 2001 law, costs and benefits generated by the intervention, the effectiveness of the gesture on the control spermograms, the satisfaction of the patients by a telephone questionnaire. RESULTS: The mean age was 41.3 years. The second consultation was carried out in 91 % of the cases but the reflection period was not respected in 24 % of the cases. Written consent was signed in 89 % of cases. Vasectomy was performed on an outpatient basis in 73 % of cases, under local anaesthesia in 6.7 % of cases. The average cost per patient was 660.63 euros for an average gain of 524.50 euros, a loss of 136.13 euros. On the control spermogram, 54.3 % were azoosperms but the 3-month delay was not observed in 23 % of them. No patients expressed regret after surgery. CONCLUSION: The recommendations of the 2001 law were not systematically followed. This lack of standardization of practices, potential reflection of a lack of interest, is to be highlighted with the extra cost generated. The revaluation of the act should be integrated into the reflection of improvement of male sterilization practices. LEVEL OF PROOF: 4.

Determining the success of vasectomy.

OBJECTIVES: To examine patient compliance, significance of rare nonmotile sperm (RNMS) and to determine the timing and number of semen analyses required to confirm sterility. PATIENTS AND METHODS: From November 2001 to November 2004, 436 consecutive primary vasectomies were performed by one surgeon. All patients were instructed to submit two initial semen specimens for analysis (2 and 3 months after vasectomy) and additional samples (at 1-month intervals) if sperm were identified on the initial and subsequent analyses. RESULTS: A quarter of the patients submitted no semen specimens and only 21% followed the full instructions to provide two consecutive negative semen analyses. Three-quarters of the patients provided a semen specimen at 8 weeks after vasectomy; of these, 75% were azoospermic and 25% contained sperm. At 12 weeks after vasectomy half the patients provided a semen specimen; of these, 91% were azoospermic and 9% contained sperm. Of the 83 patients with semen containing sperm at 8 weeks, 80 had RNMS and three had rare motile sperm (one of whom subsequently proved to have vasectomy failure). Of the 80 patients with RNMS, at 3, 4, 5, 6, 8, 10 and 11 months, 65, four, three, four, two, one and one, respectively were azoospermic. CONCLUSIONS: The present results indicate that many patients are not compliant with the protocol after vasectomy. Provided patients have been adequately counselled, we think that one negative semen analysis at 3 months or one with RNMS at 2 months may be adequate to determine the success of vasectomy. This should reduce the number of semen analyses, including reducing the number of men who must undergo repeat testing, without sacrificing the accuracy of determining paternity. Simplifying the follow-up after vasectomy is important; not only would it be cost-effective but it may also improve patient compliance.

Measuring mouse sperm parameters using a particle counter and sperm quality analyzer: a simple and inexpensive method.

This study examined a method for analyzing the count, motility, and morphology of mouse epididymal sperm, optimizing the diluent, incubation time, sample concentration, and temperature, using a particle counter (CDA-500) to count and size sperm and a sperm quality analyzer (SQA-IIC) to measure sperm motility, quantified as the sperm motility index (SMI). The optimal conditions consisted of a 30-min incubation in D-MEM (Dulbecco's modified Eagle's medium; considering cost and availability) at 37 degrees C, with 5 x 10(6)cells mL(-1) in the original solution. Furthermore, the influence of formalin fixation, and the correlation between the automated counter and a manual method were investigated. The sample fixation had no marked effect on the sperm count or morphology assessment. A linear correlation was observed between the manual and automated methods (y=0.920x +0.276; r(2)=0.571; p<0.001; range: (3-6) x 10(6)). The suitability of the proposed method was confirmed using spermatozoa prepared from mice treated with the reproductive toxin diethylstilbestrol (DES). Using sperm from the cauda epididymidis on one side per mouse, we confirmed that measurement of these sperm parameters using the two devices was simple, rapid, inexpensive, and reproducible.

Halosperm is an easy, available, and cost-effective alternative for determining sperm DNA fragmentation.

The characteristics of Halosperm make this kit a reasonable alternative to allow basic and clinical research on sperm DNA fragmentation in any basic laboratory around the world.