With the recent development in science and technology, the advancement in in vitro fertilization to treat infertility has been one of the most revolutionary advances. However, the success rate of the IVF process depends on the efficiency of each step of the process. The physical tools used to enhance the process continue to change. Microfluidics technology is an emerging technology being used in multiple biological applications for the miniaturization and specification of laboratory techniques. Technology is used along with IVF to enhance the outcome by facilitating every step of the process. Microfluidics can be used to handle gametes, culture embryos, cryopreservation, and for many other applications. This review will highlight the applications of microfluidics in different stages of IVF, including the handling of gametes, sperm collection and isolation, sperm sorting, embryo culture, cryopreservation and the fabrication process of microfluidics, focusing on the benefits and shortcomings of these applications.

Introduction: Brucellosis is a highly contagious zoonotic disease caused by ingestion of unpasteurized milk or undercooked meat from infected animals or close contact with their secretions. Subject and Methods:Sero-prevalence of brucellosis in pregnant women was conducted for the first time in Kathmandu, Nepal. A total of 80 sera samples were collected from the pregnant women visiting Kathmandu Model Hospital. The patients were categorized on the basis of age, trimester and ethnic groups. The sera samples were tested by ELISA method. Results:The sero-prevalence of brucellosis among pregnant women was found to be 11.25%. Madhesi ethnic group showed the highest (16.66%) seropositivity rates followed by Janajati (11.53%) and the lowest was in Brahmin (8.33%) ethnic group. Similarly, the age group 31-35 years showed highest prevalence (29.41%) followed by the age group 26-30 years (13.33%). There is absence of seropositivity among the age group 16-20 years and 21-25 years. The highest sero-prevalence rate (12.76%) was found in the third trimester followed by first trimester (10%) and the lowest was in second trimester (8.69%). About 3% of them consume raw milk directly from milking animals which is one of the risk factor of brucellosis in pregnant women. Conclusion:The prevalence was found to be high in pregnant women and ELISA was a sensitive and specific test for the detection of IgG antibodies against Brucella.

A fluorometric assay for single substrate-based glucose and lactate measurements was demonstrated by adopting an already established protocol for glucose and by optimizing pH, enzyme concentration, substrate concentration, and types of buffers for lactate. Linear calibration curves for glucose and lactate concentrations from 1 to 100 μM were obtained with correlation coefficients (R 2) of 0.94 and 0.98, respectively. First, with the optimized protocol, single embryo quality was successfully evaluated. Three different initial stages of embryos (n = 58) were cultured for 24 h, and glycolytic activities were calculated by measuring amounts of glucose consumption and lactate production. Results showed that embryos cultured at a later stage had lower glycolytic activities, implying more developmental activities. Second, glucose and lactate concentrations in blood plasma of diet-induced obese (DIO) mice were measured. Levels of both glucose and lactate in DIO mice were higher than those in normal mice by 2.15 and 3.8 mmol/L, respectively (both p < 0.001). Finally, clinical serum samples were analyzed and categorized into three groups based on their measured glucose concentrations: normal (4.73 ± 0.29 mmol/L), prediabetic (6.49 ± 0.13 mmol/L), and diabetic (11.34 ± 1.36 mmol/L) (p < 0.05). Collectively, this developed technique can be used to select a high-quality embryo for transfer as well as to measure glucose and lactate levels in other biological samples.

In this study, in vitro preimplantation embryo culture media especially for outbred stock mice (Institute of Cancer Research (ICR)) were optimized with different concentrations of ethylenediaminetetraacetic acid (EDTA). A plot with embryo development rates against EDTA concentrations ranging from 0 to 500 µM showed a unique pattern with two characteristic peaks. Two hundred micromolar was adopted as an optimal concentration of EDTA. The optimized media were also evaluated with two culture systems: conventional large volume culture system (1 ml) and micro-droplet culture system. In the conventional large volume culture system, the blastocyst development rates were compared among three different media (F-10, KSOM and KSOM with the optimized 200 µM EDTA). The rates were 0.4%, 16.7% and 57.6%, respectively. The development rates for the micro-droplet (10 µl) culture system were 73.9%. In conclusion, 200 µM EDTA concentration in 10 µl droplets in the KSOM medium was found as the most suitable culture conditions for ICR mouse embryos, as the blastocyst development rate was higher in the micro-droplet culture system than in the traditional conventional large volume culture system.