GM501 Gradient 100% is an isotonic solution for semen preparation with a density of approximately 1.12 g/ml.

GM501 Gradient 100% can be used in combination with IUI, IVF and ICSI.

Product facts and notices

Composition

• GM501 Gradient 100 % consists of silane-coated colloidal silica particles suspended in HEPES-buffered medium.

Product specifications and quality control

• All raw materials are of highest available purity (European Pharmacopoeia and/or USP standard) if applicable.
• A certificate of analysis is available for each batch upon request from our website with respective lot number.
• The MSDS for GM501 Gradient 100 % is available upon request and can also be downloaded from our website.
• GM501 Gradient 100 % is manufactured according to the following specifications:

Instruction for use:

• We advice to produce a 2-phasic gradient system (45% and 90%). You may prefer a different mixing ratio (e.g. 40% and 80%) or a multi layer gradient (45%-70%-90%).
• Mix 9 shares of GM501 Gradient 100 % with 1 share of washing medium to produce the 90% gradient.
• Mix 4.5 shares of GM501 Gradient 100 % with 5.5 shares of washing medium to produce the 45 % gradient.

Note: Gradients should be prepared and repacked under sterile conditions (e.g. LAF-bench, ISO Class 5). For optimal results prepare the gradient media maximum 24 hours prior to use. Mix well after diluting the GM501 Gradient 100 %.

Calculation of G-Forces:

• The g-force of your centrifuge can be calculated using this formula:

g = 1.118 x r x rpm² or rpm = Square root {g/ (1.118 x r)}

r = radius of centrifuge in mm
rpm = rotations per minute/1000

Example 1
r = 150 mm
rpm = 1200 rotations per minute
g = 1.118 x 150 x 1.44 = 242 g

Example 2
r = 150 mm
g = 300 g

rpm = SQR {300/ (1.118 x 150)} = 1.33
rpm = 1330 rotations per minute

Instruction for use with frozen semen samples:

1. Before use warm all components of the system and the samples to 37°C or to room temperature.
2. Mix the prepared density gradients by 5 bottle inversions before use.
3. Pipet 2.5 ml of the lower density gradient (45 %) into a sterile disposable centrifuge tube.
4. Using a 3 cc syringe with a 21 G needle, layer 2.5 ml of the higher density gradient (90 %) under the lower density gradient (45 %) free of air bubbles. Take care that the two layers are distinctly separated. This is done by placing the tip of the needle at the bottom of the centrifuge tube and slowly dispensing the higher density gradient. These two layers of density are stable for about two hours.
5. Gently place 2.5 ml of thawed semen onto the upper layer using a transfer pipette or syringe.
6. Centrifuge at 350-400 g for 15-18 minutes. In case no pellet is visible after this step, centrifuge for another 3 minutes.
7. Aspirate the supernatant.
8. Using a syringe, resuspend the pellet with 2-3 ml of fresh washing medium.
9. Centrifuge at 300 g for 8-10 minutes. In case you want to gain higher sperm concentrations it is advisable to centri­fuge for the whole 10 minutes.
10. Aspirate the supernatant and repeat the last two steps.
11. Finally remove the remaining liquid to have the pellet resuspended again in the desired amount for the use with the subsequent procedure of assisted reproductive medicine (e.g. IVF, ICSI, IUI).

Precautions and warnings:

• All human organic material should be considered potentially infections. Therefore, handle all specimens as if capable of transmitting HIV or hepatitis.
• Always wear protective clothing when handling specimens.
• Always work under strict hygienic conditions (ISO 5 environment, e.g. LAF-bench) to avoid possible contamination.
• Only for the intended use.

Pre-use checks:

• Do not use the product if bottle, seal of the container or package is opened or defective when the product is delivered.
• Do not use the product if it becomes discoloured, cloudy or shows any evidence of microbial contamination.