NidaCon International AB


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Frequently asked questions

  • What should I use for diluting PureSperm 100?

    PureSperm Buffer is the best media to use for dilution of PureSperm100. It has been optimised and ionically balanced to match PureSperm.

    Is there any human serum albumin in PureSperm 100?

    Research in our laboratory showed that the yield of motile sperm recovered from the gradient was approximately the same, regardless of whether or not the gradient layers contained human serum albumin. Therefore, we have not included any protein in PureSperm 100. The colloid in these products eliminates aggregation of sperm and reduces their sticking to the centrifuge tube. However, human serum albumin must be added to the “wash” solution, as provided in PureSperm Wash.

    Are antibiotics included in PureSperm 100?

    No, PureSperm 100 does not contain antibiotics, for the following reasons: 1. The gradient will remove most, if not all, of the bacterial contamination present in the ejaculate, provided that the retrieval of the sperm pellet is carried out according to the instructions given in the package insert. 2. Antibiotics commonly used in cell culture media are toxic to sperm.

    What should be used for washing the sperm pellet after the PureSperm 100 gradient centrifugation?

    We recommend PureSperm Wash for washing the sperm pellet after gradient centrifugation. The ionic balance of PureSperm Wash lies between that of the gradient and commonly used fertilisation media and, therefore, will not cause premature hyperactivation of the sperm. PureSperm Wash is also ready-to-use and contains an appropriate level of human serum albumin.

    Why do I need to buy a separate product, such as PureSperm Buffer to dilute PureSperm 100? Could I use PureSperm Wash instead?

    Yes, you can use PureSperm Wash to dilute PureSperm. However, a comparison in our laboratories has shown that you will obtain a better yield of motile sperm from using PureSperm Buffer which has been optimised for this purpose.

    Do we need to incubate PS100 and PS40/80 in 6% CO², 37°C before use?

    The most important to avoid is temperature fluctuations for the sperm. The ejaculate is approximately around room temperature or slightly higher. Therefore we recommend equilibrating PureSperm to room temperature, not more. The preparation is also done at room temperature and therefore the temperature is still maintained.

  • What diluent has been added to PureSperm to produce PureSperm 40 and PureSperm 80?

    PureSperm 40 and PureSperm 80 are optimised products in their own right; they are not diluted PureSperm 100. However, the colloid, salts and buffering capacity of these products have been optimised to provide the ideal “40%” and “80%” gradient layers.

    Is there any human serum albumin in PureSperm 40 and PureSperm 80?

    Research in our laboratory showed that the yield of motile sperm recovered from the gradient was approximately the same, regardless of whether or not the gradient layers contained human serum albumin. Therefore, we have not included any protein in PureSperm 40 and PureSperm 80. The colloid in these products eliminates aggregation of sperm and reduces their sticking to the centrifuge tube. However, human serum albumin must be added to the “wash” solution, as provided in PureSperm Wash.

    Are antibiotics included in PureSperm 40 and PureSperm 80?

    No, PureSperm 40 and PureSperm 80 do not contain antibiotics, for the following reasons: 1. The gradient will remove most, if not all, of the bacterial contamination present in the ejaculate, provided that the retrieval of the sperm pellet is carried out according to the instructions given in the package insert. 2. Antibiotics commonly used in cell culture media are toxic to sperm.

    What is the buffer component of PureSperm 40 and PureSperm 80?

    PureSperm 40 and PureSperm 80 contain Hepes buffer to avoid fluctuations in pH which might be harmful to sperm.

    What should be used for washing the sperm pellet after the PureSperm 40 and PureSperm 80 gradient centrifugation?

    We recommend PureSperm Wash for washing the sperm pellet after gradient centrifugation. The ionic balance of PureSperm Wash lies between that of the gradient and commonly used fertilisation media and, therefore, will not cause premature hyperactivation of the sperm. PureSperm Wash is also ready-to-use and contains an appropriate level of human serum albumin.

    Do we need to incubate PS100 and PS40/80 in 6% CO², 37°C before use?

    The most important to avoid is temperature fluctuations for the sperm. The ejaculate is approximately around room temperature or slightly higher. Therefore we recommend equilibrating PureSperm to room temperature, not more. The preparation is also done at room temperature and therefore the temperature is still maintained.

  • How do I use PureSperm Buffer?

    PureSperm Buffer is used to dilute PureSperm 100 to obtain the layers of different densities required for a discontinuous density gradient. PureSperm Buffer has been carefully formulated to give optimal results when used according to the manufacturer´s instructions. It is ready-to-use, no additives being required.

    Why do I need to use PureSperm Buffer to dilute PureSperm 100?

    A comparison has shown that you will obtain a better yield of motile sperm from using PureSperm Buffer for diluting PureSperm than non-optimised products. commercially available culture media, washing media or salt solutions such as Earles´Balanced Salt Solution have not been optimised for this purpose. Furthermore, salt solutions such as Earles, requires other ingredients to be added before use, thus increasing the cost.

    Why do I need to buy a separate product, such as PureSperm Buffer to dilute PureSperm 100? Could I use PureSperm Wash instead?

    Yes, you can use PureSperm Wash to dilute PureSperm 100. However, a comparison in our laboratories has shown that you will obtain a better yield of motile sperm from using PureSperm Buffer which has been optimised for this purpose.

    Should I equilibrate PureSperm Buffer before use?

    If the bottle of PureSperm Buffer has been opened previously and then stored in the refrigerator, it should be brought up to room temperature before use to avoid cold-shocking the sperm. It is not necessary to equilibrate PureSperm Buffer in the CO² incubator before use.

    Are there any antibiotics in PureSperm Buffer ?

    No, PureSperm Buffer does not contain antibiotics. Many commonly-used antibiotics are toxic to sperm. Furthermore, antibiotics such as penicillin, have a very short half-life in solution. When used according to the manufacturer´s instructions, the PureSperm density gradient will enable an uncontaminated sperm preparation to be obtained from the semen sample.

  • When should PureSperm Wash be used?

    We recommend PureSperm Wash for washing the sperm pellet after centrifugation using a PureSperm density gradient. The ionic balance of PureSperm Wash lies between that of the gradient and commonly used fertilisation media and, therefore, will not cause premature hyperactivation of the sperm. PureSperm Wash is also ready-to-use and contains an appropriate level of human serum albumin.

    What are the constituents of PureSperm Wash?

    PureSperm Wash is an isotonic (290-300 mOsm/kg H2O), HEPES-buffered salt solution containing glucose, EDTA and human serum albumin.

    Is there any protein in PureSperm Wash?

    Research in our laboratory showed that human serum albumin must be added to the PureSperm Wash to prevent sperm sticking to plastic-ware and microscope slides.

    Are antibiotics included in PureSperm Wash?

    No, PureSperm Wash does not contain antibiotics, for the following reasons: 1. Antibiotics commonly used in cell culture media are toxic to sperm. 2. The gradient will remove most, if not all, of the bacterial contamination present in the ejaculate, provided that the retrieval of the sperm pellet is carried out according to the instructions given in the package insert.

    Should PureSperm Wash be refrigerated?

    PureSperm Wash is supplied as a sterile solution which does not need refrigerated storage in the unopened bottle. However, after opening under sterile conditions, PureSperm Wash should be kept at 2-8°C.

  • What do I need to perform the sperm preparation using Speedikit?

    You only need a centrifuge and some sterile pipettes. PureSperm Speedikit includes the prefilled tubes for both separation and washing of the sample.

    Do I need an incubator for performing a sperm preparation with Speedikit?

    No, the whole procedure is performed at room temperature.

  • I have a very viscous sample; will it go through the ProInsert?

    It will go through but you can improve the outcome by treating your viscous sample with PureSperm Buffer before use. Dilute your sample with PureSperm Buffer 1+3 incubate in 37°C for 15-30 minutes, mix and it’s ready for the density gradient preparation. You can find more information on our website.

  • Can we use the same protocol for SpermCatch as we have used for PVP products?

    Yes, you can use SpermCatch in the same way as products containing PVP.

  • Why is NidOil packed in amber bottles?

    There have been several reports of other commercially available paraffin oils becoming embryo–toxic after exposure to light on the laboratory bench. Therefore, NidOil is packed in amber bottles as a precaution against light-induced changes to the product.

    Should I equilibrate NidOil in the CO² incubator before use?

    Yes, NidOil should be equilibrated in the same way as the culture medium before use to avoid differences in temperature and gaseous content between the components of the culture system.

  • What survival rates should we expect after freezing/thawing?

    The number of spermcells surviving a freezing procedure can vary significantly between samples. Analyses on freezing/thawing donor samples however has shown average survival rates of between 65-70%. (“Survival” has been defined as the number of motile spermcells after thawing compared to the number of motile spermcells before freezing.)

    Can I use Sperm CryoProtec  for TESE?

    Yes you can, you don’t have to add anything it’s as easy as using Sperm CryoProtec  with ejaculated sperm.

    Do I need to add egg-yolk to Sperm CryoProtec before use?

    No you don’t . Sperm CryoProtec is a ready to use freezing medium which doesn’t require any additives. Just follow the instructions and your sperm will be safely frozen and thawed.

    Does Sperm CryoProtec contain antibiotics?

    No, none of our products contains any antibiotics. Adding antibiotics would significantly reduce the shelf life of our products and could “mask” post-sterilisation contamination of the product

  • How many cells do I need to count?

    We recommend that you count 200 sperm to get an accurate classification. The 100x objective with immersion oil will give you a very clear picture of the stained versus unstained sperm cells.

    If the sperm is only coloured at the neck is classifies as dead or alive?

    It’s classified as alive.

  • Can Blastocysts cultured in any culture media be vitrified with your kits?

    VitriBlast /ThermoBlast kit can be used to freeze blastocysts that have been cultured in any culture media.

    What device should I use with the VitriBlast kit?

    Any device can be used but our general recommendation is to use a closed-system.

    Is DMSO more harmful compared to other cryoprotectants?

    No, actually in general, DMSO is one of the least harmful organic solvents. It has a very low toxicity , it is not genotoxic, not mutagenic or carcinogenic.

    It is a FDA approved drug to treat interstitial cystitis, but has been (and is still) used widely to treat various diseases including arthritis, local pains, inflammation, infections (herpes), autoimmun problems and also gastric ulcer.

    In embryology, DMSO was reported to protect membranes better than other cryoprotectants; less toxic than propylene glycol; and is the standard component of the most successful vitrification mixtures both in humans and domestic animals.

  • Can Blastocysts cultured in any culture media be vitrified with your kits?

    VitriBlast /ThermoBlast kit can be used to freeze blastocysts that have been cultured in any culture media.

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