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Tips and Tricks

September 2017

September 2017 Tips & Tricks

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).

 

your best option will be to acquire the NidOil 400 Kit, (4 x 100 ml).

June 2017

June 2017 Tips & Tricks

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.

May 2017

May 2017 Tips & Tricks

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.

Image credit: Genome Research Limited

April 2017

April 2017 Tips & Tricks

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 click here
or
by using the calculator in our web-site, just add you data and the calculator will do the rest. click here.


March 2017

March 2017 Tips & Tricks

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.

Sperm VitalStain

February 2017

February 2017 Tips & Tricks

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.
Study

January 2017

January 2017 Tips & Tricks

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: 

Use this link Video to see what is present in the seminal plasma, the layers and the rafts in between and what the final preparation can look like.

December 2016

December 2016 Tips & Tricks

TIPS AND TRICKS 2016

  • March 2016
    VISCOUS SEMEN SAMPLES
    Obtaining satisfactory sperm yield from highly viscous semen samples remains a major problem for laboratories providing services to assisted reproduction programs.
  • April 2016
    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.
  • May 2016
    LOSS 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.
  • August 2016
    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.
  • September 2016
    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:
  • October 2016
    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.
  • November 2016
    SINCE YOU STORE THE ADDITIVES IN THE REFRIGERATOR, THE DMSOWILL 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.


November 2016

November 2016 Tips & Tricks

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.

October 2016

October 2016 Tips & Tricks

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!


September 2016

September 2016 Tips & Tricks

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.

Wash+PS100

August 2016

August 2016 Tips & Tricks

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 download it

July 2016

July 2016 Tips & Tricks

TAKE INTO ACCOUNT THIS SUMMER

Tips and Tricks:

Plan ahead remembering that Nidacon will be closed from July 18th to July 31st

If you want to spend an unforgettable summer, here are the top 10 tips for enjoying Swedish summer:
Top 10 Tips

June 2016

June 2016 Tips & Tricks

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.

May 2016

May 2016 Tips & Tricks

LOSS 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 SpermCryoProtec with your sample but don’t mix for longer than 5 minutes since glycerol is toxic to cells at RT.


April 2016

April 2016 Tips & Tricks

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.

 


March 2016

March 2016 Tips & Tricks

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).



ADDRESS

Flöjelbergsgatan 16B Mölndal 43 137 – Sweden
Phone: +46-31-703 06 30
Fax: +46-31-40 54 15
Website: http://nidacon.com
Email: contact@nidacon.com

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