Instructions
Instructions
Container reception
Container reception
Once the sample has been used, CEIFER Biobanco must be contacted to proceed with the return of the cryocontainer, by e-mail to secretaria@ceifer.com Please carry out this operation no later than three days after receiving it. When returning the cryocontainer, use the safety seal designed to close the packaging
Sample preparation
Sample preparation
Whole Semen
The ejaculate (whole semen) is diluted with cryoprotectant medium (Freezing Medium – TEST yolk buffer from IrvineScientific) at 1:1 ratio. Then it is stored into straws and frozen.
- Culture Media: Sil-Select STOCK 100 % TM /Ferticult TM Flushing/Rincage
- Pasteur pipettes ® sterile fine tip (Deltalab. Ref. 210024)
- Centrifuge (preferentemente de cabezal basculante)
- Cultivation oven at 37 ° C
- The chosen products were chosen because they were used in our laboratory.
- There are other products on the market that can replace the recommended ones.
- CEIFER Biobanco does not assume responsibility for the results that each laboratory may obtain with this method. Our responsibility is to provide an adequate number of motile sperm after thawing of the straws.
- Gradient 80 %: 2 mL Ferticul + 8 mL Sil-Select
- Gradient 40 %: 6 mL Ferticul + 4 mL Sil-Select
- They can be kept for several weeks at 2-8 ° C, if prepared under sterile conditions.
- It is advisable to leave 1.5 mL of each gradient at 37 ° C, from the day before use.
- Pour 1.5 ml of the 40% gradient with a Pasteur Pipette into a Falcon tube.
- Pour 1.5 ml of the 80% gradient into the bottom of the same tube with a Pasteur Pipette slowly and through the 40% gradient previously. </ li >
- Keep at 37 ° C until use.
- Thaw 3 straws, 10 minutes, in an oven at 37 ° C.
- Dry the straws externally, as condensation will have formed.
- Cut the distal end of the straws, insert into a sterile tube and cut the proximal end. The content will fall into the tube.
- Add 4.5 mL of Ferticult Flushing / Rincage medium (previously tempered at 37 ° C) drop by drop, shaking very slowly.
- Total 6 mL will be obtained (4.5 mL of Ferticult + 1.5 mL of semen).
Whole Semen
- Culture Media: Sil-Select STOCK 100 % TM /Ferticult TM Flushing/Rincage
- Pasteur pipettes ® sterile fine tip (Deltalab. Ref. 210024)
- Centrifuge (preferentemente de cabezal basculante)
- Cultivation oven at 37 ° C
- The chosen products were chosen because they were used in our laboratory.
- There are other products on the market that can replace the recommended ones.
- CEIFER Biobanco does not assume responsibility for the results that each laboratory may obtain with this method. Our responsibility is to provide an adequate number of motile sperm after thawing of the straws.
- Gradient 80 %: 2 mL Ferticul + 8 mL Sil-Select
- Gradient 40 %: 6 mL Ferticul + 4 mL Sil-Select
- They can be kept for several weeks at 2-8 ° C, if prepared under sterile conditions.
- It is advisable to leave 1.5 mL of each gradient at 37 ° C, from the day before use.
- Pour 1.5 ml of the 40% gradient with a Pasteur Pipette into a Falcon tube.
- Pour 1.5 ml of the 80% gradient into the bottom of the same tube with a Pasteur Pipette slowly and through the 40% gradient previously. </ li >
- Keep at 37 ° C until use.
- Thaw 3 straws, 10 minutes, in an oven at 37 ° C.
- Dry the straws externally, as condensation will have formed.
- Cut the distal end of the straws, insert into a sterile tube and cut the proximal end. The content will fall into the tube.
- Add 4.5 mL of Ferticult Flushing / Rincage medium (previously tempered at 37 ° C) drop by drop, shaking very slowly.
- Total 6 mL will be obtained (4.5 mL of Ferticult + 1.5 mL of semen).
Ready-use Semen
The ejaculate is processed following a technique for the recovery of motile spermatozoa through density gradients, washing out and dilution (1:3 ratio) using ‘Sperm Freeze’ medium (FertiPro). Then it is stored into straws and frozen.
- Defrost 1 straw, 10 minutes, in an oven at 37 ° C.
- Dry the straw externally, as condensation will have formed.
- Cut the distal end of the straw, insert it into a sterile tube and cut the proximal end. The content will fall into the tube.
- It is recommended to wash the thawed sample for the removal of cryoprotectants with the usual protocol of your laboratory.
Readyuse Semen
- Defrost 1 straw, 10 minutes, in an oven at 37 ° C.
- Dry the straw externally, as condensation will have formed.
- Cut the distal end of the straw, insert it into a sterile tube and cut the proximal end. The content will fall into the tube.
- It is recommended to wash the thawed sample for the removal of cryoprotectants with the usual protocol of your laboratory.