NidaCon International AB


Flöjelbergsgatan 16B
431 37 Mölndal


Map

+46 31 703 06 30

Contact us


Contact

Fill in the form


Tips and tricks

  • FREEZING PROTOCOL FOR SPERMCRYOPROTEC
    COULD WE MAKE IT EASIER FOR THE EMBRYOLOGIST, WHILE MAINTAINING THE HIGH SURVIVAL RATE?

    Flexibility is luxury today and something that you don’t have a lot of in an IVF-lab. Exact time protocols to follow, patients arriving at a certain time etc., can sometimes be quite stressful.
    After a lot of testing and help from some of our customers (thank you Akademiska in Uppsala), 2 variables are now changed:
    Tips and Tricks

    1. The first is the incubation time in the fridge after loading the straws.

    The new range is now 10-60 minutes instead of earlier 30-60 minutes. If you look at the results when incubating for 60 minutes compared to 10, you can see a slight increase when incubated for 60 minutes. The big difference is however between not incubating and incubating for 10 minutes.

    2. The second change is the time on the CryoFloater in LN2

    The new recommendation is 10-30 minutes instead of only 30 minutes. We have again performed a lot of tests, both at Nidacon and with the help of Akademiska Hospital in Uppsala, Sweden. If you measure the temperature on the floater, it does go down quite fast and 10 minutes is more than enough but, in order to make it more flexible, we now recommend the range 10-30. No difference in result with regards to survival rate has been found in our tests between 10 and 30 minutes but it will give you the chance to take the coffee you so desperately need!

  • How does the centrifuge rotor affect the yield from a PureSperm gradient?

    The centrifuge rotor can have a big influence on the outcome of your density gradient.
    Centrifugation using a swing-out rotor is more likely to give a better yield of motile sperm than a fixed rotor centrifuge.

    With a swing-out rotor, the tubes are vertical when they are placed in the centrifuge, but when the rotor starts to spin, the tubes swing out to be horizontal, so the sperm pass directly through the gradient layers, in the direction of the centrifugal force, and collect in a pellet at the bottom of the tube (as illustrated in the figure).

    In contrast, the sperm that pass through the gradient in a fixed-rotor centrifuge are not in a tight pellet, rather the pellet is more of a smear across the side/ bottom of the conical centrifuge tube.

    This is because the direction of movement through the gradient is not the same as the direction of the centrifugal force, so it is less efficient.

    As it is harder to retrieve sperm from a more diffuse pellet than a tight pellet, centrifugation using a swing-out rotor is more likely to give a better yield of motile sperm.

  • TEMPERATURE AND SPERM DENSITY GRADIENT PREPARATION

    Spermatozoa are made to fertilize eggs. However, spermatozoa are not ready to interact with eggs when ejaculated. They need to undergo a physiological change called capacitation before penetrating the egg.
    Capacitation is a temperature dependent phenomenon; variation in the incubation temperature could cause an alteration in some events associated with this process.Sperm incubation a room temperature produces a temporary blockage of capacitation related events. When maintained under these conditions, cells are in a “quiescent” state (non-capacitated), retaining their functional properties that can be expressed at body temperature of 37°C (1)The rise in temperature when spermatozoa enter the female reproductive tract could act as a trigger for the activation of spermatozoa, making them hyperactive (Gallup,2009). This hypothesis possibly explains the diminished survival of spermatozoa at 37°C compared with lower temperatures.

    Tips and Tricks:

    We recommend density gradient to prepare the semen samples and performed at room temperature. Remember, the best is to avoid temperature fluctuation and therefore, the best way to prepare and store the sample, is at room temperature.

    Ref:

    Influence of temperature and sperm preparation on the quality of spermatozoa.
    Thijssen, Klerkx, Huyser, Bosmans, Campo, Ombelet.
    Reprod Biomed Online. 2014 Apr;28(4):436-42. doi: 10.1016/j.rbmo.2013.12.005. Epub 2014 Jan 14.

  • How to optimize your work


    Proper handling of sperm contributes to improved fertilization rates. An undisputed fact.

    Always prepare your sperm sample by performing a two layer PureSperm-gradient. This is equally important if you are to use the sperm fresh or if your sperm samples are to be frozen.

    By proper preparation, freezing, and thawing of the sperm, you optimize both survival and motility.

    If you find this procedure complicated, you can choose to use the ProInsert to simplify it and at the same time minimise the risk for mistakes.

    Tips and Tricks:

    Attend our hands-on training workshop at no cost. The first workshop 2018 will take place at Nidacon facilities in
    Gothenburg -Sweden on May 04.
    Info contact@nidacon.com
    Agenda:

    09.00   Nidacon introduction
    09.15   Product presentation
    10.00   Hands-on
    12.00   Lunch
    13.00   Hands-on
    14.30   Coffee
    15.00   Latest research: What actually happens to the sperm in the lab
    16.00-16.30 Summary, discussions

  • Cryopreservation of Mammalian Gametes and Embryos Methods and Protocols
    Editors: Nagy, Zsolt Peter, Varghese, Alex C., Agarwal, Ashok (Eds.)

    This volume provides a comprehensive and technical presentation of numerous aspects of reproductive cell tissue cryopreservation, and presents readers with current procedures and detailed discussions of novel techniques and the latest innovations.

    • Includes cutting-edge methods and protocols
    • Provides step-by-step detail essential for reproducible results
    • Contains key notes and implementation advice from the experts

    www.springer.com

    Tips and Tricks:

    Appendix G: Vitrification of blastocysts using VitriBlast and ThermoBlast: Nidacon
    Anna Niläng, Thorir Hardarsson, Ann-Sofie Forsberg, Tetsunori Mukaida, and Paul V. Holmes.

    Abstract

    This appendix describes the vitrification of blastocysts using VitriBlast and ThermoBlast, from Nidacon. The technique used and the reason for not including DMSO in the medium at the production stage, but including it separately in the kit, and the importance of collapsing the blastocyst prior to vitrification will be explained and described.

  • HOW NIDACON REDUCE THE IMPACT OF TRANSPORTS AND THEREBY ADDRESSING CLIMATE CHANGE

    At Nidacon, we want to contribute to a sustainable world by taking responsibility for our impact on society, the environment and the people of our planet.

    We believe that through strategic work with CSR (Corporate Social Responsibility), we can strengthen our ability to identify and prevent potential harmful effects, but above all, create valuable opportunities for our own business and society as a whole.

    Tips and Tricks:

    Our main logistics partner is UPS and since we transport products all over the world we compensate for the carbon emissions caused by these transports through being part of the UPS Carbon Neutral Program.

    This means that we support emissions reduction projects that help mitigate the climate impact every tonne of carbon dioxide a parcel from Nidacon produces in transportation.

  • THE IDEAL SPERM SEPARATION TECHNIQUE FOR ART

    Under in-vivo conditions, potentially fertile spermatozoa are separated from immotile spermatozoa, cellular debris, and seminal plasma in the female tract by active migration through the cervical mucus.

    Sperm quality also plays a very prominent role in achieving a pregnancy through ART, and a variety of procedures exist for selecting better quality sperm for egg fertilization, such as Density Gradient and Swim-up.

    Tips and Tricks:

    The ideal sperm separation technique should:

    • Be quick, easy and cost-effective.
    • Concentrate as many normal, motile spermatozoa as possible.
    • Separate out the morphologically abnormal, the immature and the senescent sperm.
    • Eliminate macrophages, leukocytes, cellular detritus, bacteria and virus particles.
    • Remove toxic or bioactive substances, including decapacitation factors and reactive oxygen species (ROS).
    • Select sperm with longer telomeres and eliminate sperm with DNA fragmentation.
    • The Differential Density Gradient Technique is a gentle method of separating sperm from semen and is accepted as the state-of-the-art technology by reproductive specialists and scientists.

  • OILS BECOMING EMBRYOTOXIC AFTER EXPOSURE TO LIGHT

    We know that mineral oil can damage oocytes and embryos because of toxic contamination or deterioration of the quality. The quality of the oil is of high importance and requires high standards of quality testing.

    Nidoil is today tested rigorously, both the raw material and the final product.

    One of the multiple factors in which the quality of the oil can be compromised is that the paraffin oil becomes embryotoxic after exposure to light on the laboratory bench or under the microscope.

    Tips and Tricks:

    As a precaution against any light-induced changes prior to use, NidOil™ is therefore supplied in amber, screw-top 100 ml bottles.

    If you want to both save money for your lab and packaging material, your best option will be to acquire the NidOil 400 Kit, (4 x 100 ml).

  • ANTIBIOTICS AND SPERM PREPARATION

    Antibiotics are not included in our products for several reasons. Penicillin G, a commonly used antibiotic in cell culture medium, only lasts for approximately 10 days in aqueous solution, being inactivated after this time and the degradation products are cell-toxic. Furthermore, this antibiotic is ineffective against some of the bacteria most commonly found in semen. Streptomycin and gentamycin are cytotoxic. Gentamicin, in particular, has been shown to be toxic to embryos.

    For most situations Nidacon recommends using a discontinuous density gradient for preparing human sperm from semen. The gradient will remove most, if not all, of the bacterial contamination present in the ejaculate. However, many customers at some time need to use the swim-up technique and the most ideal product for this purpose is PureSperm® Wash.

    Tips and Tricks:

    PureSperm® Wash is a salt solution balanced and adjusted for the nutrition and long survival of human sperm. It functions exceedingly well for the swim-up technique.

    Since PureSperm® Wash does not contain any antibiotics and since swim-up cannot guarantee removal of bacterial contamination, it is recommended to add antibiotics when using swim-up to prepare sperm for ART. We recommend that you add Penicillin at a concentration of 100 U/ mL.

  • SELECT SPERM WITH LONGER TELOMERES AND ELIMINATE SPERM WITH DNA FRAGMENTATION

    Selection of functional sperm with less DNA damage and longer telomeres should be prerequisites for achieving optimal outcomes for ART.
    DNA fragmentation contributes to the failure of assisted reproductive technologies, and can ultimately lead to failed fertilization and increases the risk of abnormal embryo development, while telomere shortness is associated with males who are infertile ( Zhao, F. et al.)

    Tips and Tricks:

    Density Gradient Centrifugation is the optimal sperm preparation method in order to avoid DNA fragmentation and to select the sperm with the longer telomeres.

    PureSperm 40/80/90
    Simple: ready-to-use density gradient solutions, 40/80 and 90%, make work in the lab easier and they minimize the risk for mistakes.

    Flexible: a 40/80 or a 40/90 combination. Both are two-layer systems for density gradients, the 40/90 combination giving higher sperm motility, while the 40/80 gives higher sperm yield

    REF : Zhao, F. et al. Semen preparation methods and sperm telomere length: density gradient centrifugation versus the swim-up procedure.

  • ARE YOU USING THE CORRECT RPM TO ACHIEVE THE CORRECT G-FORCE?

    We want to remind you of the importance of using the correct RPM to make sure that your centrifuge uses the correct g-force.

    How to calculate the correct RPM

    Tips and Tricks:

    Is easy and simple, just download the Nidacon Nomograph for a copy
    or
    by using the calculator in our web-site, just add you data and the calculator will do the rest.

     

  • HOW TO ACHIEVE THE BEST MICROSCOPIC VIEW OF SPERM STAINED FOR VITALITY

    Use the eosin-nigrosin technique to establish the percentage of live sperm. This technique is based on the principle that a dead cell will take up the eosin and stain red.

    Sperm Vitality should be determined in semen samples that have 50% or more immotile sperm, according to the WHO laboratory manual for examination of human sperm.

    Tips and Tricks: 

    The 100x objective with immersion oil give you a very clear picture of stained versus unstained Sperm. Read more about Sperm Vitalstain.

  • POTENTIAL PROBLEMS IN EXPOSING SPERM TO PVP TO SLOW SPERM MOTILITY, PRIOR TO ICSI

    Intracytoplasmic sperm injection (ICSI) requires the capture of an individual sperm in a glass pipette for injection into the oocyte.

    Probably the most widely practised method, since it does not require special equipment, is to reduce sperm motility by placing the sperm in a viscous medium prior to nicking the tail to immobilize the sperm completely (Van Steirteghem et al., 1993).

    Previously, the only products commercially available for slowing sperm motility contained a synthetic plastic, polyvinylpyrrolidone (PVP). However, some PVP is injected into the oocyte along with the sperm.

    Tips and Tricks: 

    Physiological alternative to PVP has been sought for reducing sperm motility to facilitate capturing sperm for ICSI.

    Since hyaluronate and human serum albumin are found naturally in the mammalian reproductive tract, Nidacon has conducted a study to evaluate SpermCatch™, a viscous liquid containing hyaluronate and human serum albumin, as a potential substitute for PVP.

  • WHAT IS FOUND IN THE DIFFERENT LAYERS AFTER A DENSITY GRADIENT CENTRIFUGATION USING PURESPERM?

    Remember by doing density gradients you will remove all the unwanted factors, before ART.

    Tips and Tricks: 

    Click here to see what is present in the seminal plasma, the layers and the rafts in between and what the final preparation can look like.

  • VITRIFICATION BLASTOCIST – SINCE YOU STORE THE ADDITIVES IN THE REFRIGERATOR, THE DMSO WILL BE SOLID BELOW +18°C.

    The DMSO and Ethylene glycol (EG) additives, which are included in the VitriBlast kit, should be mixed thoroughly before use.

    Tips and Tricks: 

    You should remove the DMSO from the refrigerator at least 1 hour before use, or the day before; and let it liquify at room temperature. The DMSO needs to be above +18°C to reach liquid form.

    – If for any reason there is a shortage of time, it can be warmed in the hand or warmed in the incubator.

     

  • CRITICAL DISTANCE BETWEEN THE SEMEN SAMPLE AND THE NITROGEN SURFACE

    How do you guarantee optimal freezing temperature? When using a raft, what is the correct height of the raft?
    Studies carried out at Nidacon showed that different heights give different sperm survival rates.

    Tips and Tricks: 

    CryoFloater provides a stable raft with the correct and constant height above the nitrogen surface, thereby guaranteeing optimal freezing temperature and the best possible result.

    And it is free of charge when you order our cryoprotectant, Sperm Cryoprotec!

  • STORAGE OF SPERM SAMPLE AND DNA FRAGMENTATION

    Sperm are highly sensitive to temperature fluctuation, therefore avoid changes in temperature.
    If storage of the sperm sample is needed prior to the ART-procedure, we recommend the following:

    Tips and Tricks: 

    The best way to store the sample, is at room temperature. The DNA fragmentation will be lower when the sample is stored at room temperature as compared to storage at 37˚C.
    We recommend density gradient to prepare the semen samples and this will be done at room temperature. Remember, the best is to avoid temperature fluctuation.

  • CALIBRATE THE CENTRIFUGE

    To achieve the optimal result using a density gradient, it is critical, to make sure that your centrifuge uses the correct g-force.

    Tips and Tricks: How to calibrate your centrifuge.

    By using the following equation:

    Rpm = √[ (g/(1.118 x r)] x 10³

    r = rotational radius, the distance (mm) from the
    centre of the rotor to the bottom of a centrifuge tube
    in the bucket when raised to horizontal position
    For example; to achieve 300 x g when radius = 165
    mm the centrifuge speed must be: Rpm = √[ (300/(1.118 x 165)] x 10³ = 1275

    By using the RCF Nomograph

    Click here to read it

  • Stay updated!

    The 32nd Annual Meeting of the European Society of Human Reproduction & Embryology will this year be held at Messukeskus Expo and Convention Centre, 3-6th of July, Helsinki Finland.

    For detailed information about new products of the highest quality in the market, to discuss ideas, future plans or just have a nice conversation, you should consider the following:

    Tips and Tricks:

    Stop by our booth at location C152. We will always be willing to answer any questions you may have about our products or simply provide information on the latest. Or why not just stop by for a friendly chat.

    See you there.

  • OSS OF PROGRESSIVE MOTILITY CAUSED BY OSMOTIC SHOCK PHENOMENON

    The osmotic shock phenomenon caused by the exposure of frozen-thawed spermatozoa to isotonic conditions after a period of hypertonic exposure, is characterized by increased coiling of the sperm tail which results in loss of progressive motility (5, 10-14).

    Allow gradual osmotic adjustment.

    Tips and Tricks:

    To avoid osmotic chock for the sperm, it is important to slowly mix Sperm CryoProtec with your sample but don’t mix for longer than 5 minutes since glycerol is toxic to cells at RT.

  • PELLET RETRIEVAL WITHOUT RECONTAMINATION

    When retrieving the pellet after the gradient centrifugation, care must be taken to avoid contaminating the pellet with components of the ejaculate or upper gradient layer.

    Tips and Tricks:

    We recommend that you use a new pipette after removing most of the gradient to avoid re-contamination, for example by bacteria, or use the ProInsert device, which is mostly recommended.

  • VISCOUS SEMEN SAMPLES

    Obtaining satisfactory sperm yield from highly viscous semen samples remains a major problem for laboratories providing services to assisted reproduction programs.

    Tips and Tricks:

    Viscous samples can be treated with PureSperm®Buffer.

    You simply add PureSperm®Buffer to the ejaculate (dilution 1:4), 1 part PureSperm®Buffer and 3 parts sample (e.g. 0.5 ml PureSperm®Buffer + 1.5 ml semen sample). incubate for 15-30 minutes at 37°C and the sample is ready for preparation (preferably using a density gradient).

Subscribe to our Newsletter