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The purpose of this study was to evaluate the use of glass capillary micropipette (GCM) as a vessel for vitrification of bovine oocytes. Cumulus-oocyte complexes (COCs) were obtained from slaughter-house and washed 5 to 6 times in the washing medium (TCM-199 + 20% FBS) and randomly assigned to treatment and control group. In the first step of vitrification, COCs were exposed to first vitrification solution (VS1) (10% ethylene glycol (EG), 10% DMSO in holding medium (TCM-199 + 10% FBS: HM)) for 1 min at room temperature and then placed in VS2 solution (20% EG, 20% DMSO in HM) for 25 sec and immediately were loaded into the GCM vessel. The filled portion of GCM vessels were placed in liquid nitrogen (LN2) for 3 to 5 sec and then completely immersed and stored there. The oocytes were thawed by immersing the capillary end of the straw in 1 ml of 0.25 M sucrose in HM and gently expelling the contents. After 1 min the oocytes were transferred into 100 μl of 0.15 M sucrose in HM for another 5 min and then washed with HM twice. For examining the in vitro developmental potential of vitrified-warmed oocytes, the oocytes were placed in 50 μl droplet of maturation medium (TCM-199 + 10% FBS + 10 IU/ml PMSG + 5 IU/ml HCG) covered with paraffin oil in a CO2 incubator at 38.5ºC for 24 hrs. A high proportion of morphologically normal oocytes (90%) was recovered after vitrification-warming. The percentage of live oocytes after 24 hrs when tested with trypan blue in GCM group was 85.18%, significantly did not differ from control group (90%). The proportion of oocytes which were found to have undergone nuclear maturation did not show statistical difference between the control and GCM group (61.29% vs 40%, respectively). The results of present study demonstrated that vitrification of immature bovine oocytes in the GCM vessels and EG + DMSO solution have high survival rate.  

The ranking of colon cancer reported in the world is third in the number of patients and second in the number of deaths. An important issue in cancer studies is the changes in the mechanical properties of the cells and their effects on the cellular responses to biochemical and biophysical signals and determine injuries factors. In this study, the viscoelastic properties of grade II Ht29 colon cancer cell line were investigated with and without treatment by albendazole by Atomic Force Microscopy and Micropipette Aspiration methods. Through these two methods, the amounts of microtubules as the major factors in creating the cell mechanical properties were assessed and compared in treated and untreated cancer cells. The overall results showed the significant decreasing in the amounts of tubuline in cancer cell lines after treatment by albendazole is in agreement with its cytotoxicity. So in line with other studies, the cancer cells are face with reduction in the cell mechanical properties by reducing the amount of microtubules relative to normal cells and the albendazole strengths this property.

Background: Successful Embryo Transfer (ET) technique is a fateful step of all efforts to achieve live births fromin vitro produced embryos in assisted reproductive techniques or in knockout, transgenic or cloned animal projects. Small reproductive tract of mice and limitation of current techniques may not well satisfy the requirements for mass production of genetically modified mice. Genetic abnormalities of embryos, receptivity and uterine contractions, expulsion of embryos, blood, mucus or bacterial contamination on the transfer pipette tip, technical problems and even animal strain may affect embryo transfer outcome.Methods: In this study, two techniques of embryo transfer in mice were compared. In conventional technique the oviduct wall was punctured with a 30-gauge needle and the loaded Pasteur pipette with embryos and medium was inserted into the hole. In new technique, embryos that were loaded in modified micropipette with minimal medium were transferred directly to the oviduct by manual piston micro-pump easily.Embryo viability was evaluated considering the percentage of live healthy newborns.Results: Results of the two techniques were compared by t-test within the NPAR1WAY procedure of SAS software (ver.9.2). The average live birth rates in the novel methods was significantly higher (42.4%) than the conventional method (21.7%, p<0.05).Conclusion: In conclusion, using new embryo transfer technique improved birth rate by preventing embryos expulsion from the oviduct, saving time and easy transfer of embryos with minimum volume of medium.

cycles were cancelled in about one-third of these cases due to failed fertilization. The introduction of micromanipulation techniques such as zona drilling, partial zona dissection, and subzonal insemination reduced the incidence of failed fertilization and salvaged many IVF cycles. However, there was little improvement on fertilization and pregnancy outcome because these procedures required a relatively high number of progressive motile sperm, which in turn frequently resulted in a high incidence of polyspermy in inseminated eggs. To overcome these limitations, microinjection of a single sperm into the ooplasm (intracytoplasmic sperm injection, ICSI) was developed. ICSI resulted in a dramatic improvement in fertilization rates and revolutionized the treatment of infertile couples with severe male factor. ICSI however, is a labor intensive procedure where a highly trained embryologist uses a hydraulic micromanipulation system to pick up a single sperm and inject it under high power magnification into a human oocyte in order to fertilize the oocyte. This process is technically challenging, and consequently there is a long learning curve of up to 6 months for a trained embryologist to be able to become skillful with this procedure. We developed a robotic ICSI system featuring fast oocyte positioning, automated sperm tracking and immobilization, and adaptive oocyte injection with minimal human involvement. A cell holding device is developed to hold many oocytes into a regular pattern. A sperm tail tracking algorithm robust to the low-contrast appearance and the nonlinear fast movement of the sperm tail is developed. In addition a computer vision algorithm is developed to recognize oocyte structures for adapting injection parameters. The robotic system is able to inject 6-9 oocytes at a time markedly reducing the time the oocytes are out of the incubator and exposed to cooling or changes in PH of the culture medium that can be detrimental to embryo development. Finally, the system performs the injection the same way for each oocyte resulting in more uniform results and fewer damaged oocytes. Different from conventional ICSI setups that consist of two micromanipulators and use both a holding micropipette and an injection micropipette, our system contains only one micromanipulator and uses a single injection micropipette for the complete ICSI task.

Micropipettes or piston pipettes are used to make most volume measurements in fields such as health, chemistry, biology, pharmacy and genetics. Laboratories must ensure that results obtained using these instruments are reliable; therefore, it is necessary to calibrate micropipettes. Before the start of the calibration process, we must check the precision of measurements. The objective of this work is to compare several methods for calculating the precision of three kinds of micropipettes according to the reference value in ISO 8655-6. The medical tests will not have accurate results, if the volume of the liquid doesn’t transfer precisely by micropipettes. Thus, the physician might potentially face problems in the disease diagnosis and its control. In the NCCLS EP5-A2, there is a method to specify and assess the precision of micropipettes by using CV (Coefficient of Variation). Also there are other methods to estimate and test the CV theory, in the formal statistics texts which could be applied to assess the micropipettes precision. In this research we evaluate the precision of lab micropipettes. Three brands of micropipettes, A, B and C are assigned to measure the distilled water mass by using accurate scale which is accurate up to 10-6 to measure 50-gram weights. The experimental environment is a metrology lab which is approved by Iran Standard and Industrial Researches Organization. A technician sampled at the beginning of the experiment and then after 2 hours, the same technician repeated the sampling. Overall, each micropipette is used to measure 40 times with 10-repeat times for single measurement in 28 work days. Common statistical methods are used to estimate and test the CV. Point estimation of CV for micropipettes A, B and C were 0.50%, 0.64% and 1.56%, respectively. Furthermore, the upper limit of 95% confidence bounds for these three micropipettes using the exact method were 0.53%, 0.69% and 1.65%, respectively. Micropipette A met the ISO 8655-6 standard level, but micropipettes B and C did not. On average, measurement errors in micropipettes B and C were respectively 30% and 3.11 times more than micropipette A. By using the approach of CLS EP5-A2 and confidence interval for CV, precision of the three micropipettes were compared. Only one of them met the ISO 8655-6 standard level, but the others failed.