Frequently Asked Questions
Ethylene glycol and DMSO are added as cell membrane-permeable CPAs, and trehalose and hydroxypropylcellulose (HPC) are added as cell membrane-impermeable CPAs.
Store the opened solution in a refrigerator in consideration of safety, etc., and use it up within 2 weeks, as with other culture solutions, etc.
If 2 weeks have passed after opening the solution, or 2 weeks have not passed but the inside the vial may have been contaminated during dispensing the solution, discard it and use a new solution for hygienic reasons."
If 2 weeks have passed after opening the solution, or 2 weeks have not passed but the inside the vial may have been contaminated during dispensing the solution, discard it and use a new solution for hygienic reasons."
Do not use a heat plate. Rising the temperature of the reagent also accelerates chemical reaction. Pay attention to temperature control because it may induce toxicity of the cryoprotective substances. When using a heat plate, set it to room temperature (25 - 27°C) before use.
For both freezing and thawing reagents, it is one year from the date of manufacture for storage in a refrigerator. It is 2 years from the date of manufacture for plastic products (plates and Cryotec).
Since our reagents contain no protein or serum, the quality is not affected even at ordinary temperature. Normally, the product will be shipped without refrigerant, but if you are concerned, please contact us so that we will respond separately.
Yes, but we cannot guarantee the results equivalent to those obtained by using Cryotec products in all steps.
Yes, but we cannot guarantee the results equivalent to those obtained by using Cryotec products in all steps.
The total volume of the vial for only TS, and 300 μL for other reagents (ES, VS, DS and WS). These reagent volumes are calculated as the volumes required for proper operation, so be careful not to decrease or increase the volume.
Survival is not affected by any grade. Cryopreservation with Cryotec is only to stop the time of oocyte/embryo, so the same grade of oocyte/embryo will be obtained after thawing.
In the open method, the freezing solution containing oocytes/embryos is directly immersed in liquid nitrogen. In the closed method, containers with oocytes/embryos are sealed once and then sealed containers are placed in liquid nitrogen.
No cases of contamination between frozen samples (open method) stored in liquid nitrogen have been reported to date. Although oocytes/embryos are immersed in liquid nitrogen as a minimum volume droplet at the time of freezing, this droplet is very small and will not turn into small ice pieces. Therefore, it is said that viruses and bacteria do not disperse via liquid nitrogen.
Even in the closed method, the thawing procedure is basically the same as the open method, so the Cryotec method can be used. However, we cannot guarantee the results equivalent to those obtained by using Cryotec products in all steps.
If the survival of 30% or more of the blastomeres is confirmed, there are cases that resulted in implantation at the time of embryo transfer. From such a viewpoint, it can be determined to be survival. If 2 or more cells of 4 to 5 cells or 3 or more cells of 6 to 8 cells are alive, it can be determined to be survival, and we recommend its use for transfer.
All our reagents are highly viscous; even if bubbles are formed, they will disappear spontaneously. If you try to remove it, the liquid volume may change, so you can leave bubbles as they are even if they are formed.
We recommend magnification ratios where the circumference of each well overlaps that of the visual field of your microscope (x12-15) for general operations such as transfer of embryos, and the maximum magnification ratio (about x45-55) of your stereoscopic microscope for steps requiring observation of oocytes/embryos.
The design allows for easy identification of the surface where oocytes/embryos are placed and smooth attachment of the cover cap.
The amount of the reagent to be aspirated is the minimum necessary amount, so you may aspirate as indicated regardless of the difference in the inner diameter of the pipette. It is assumed that the appropriate size of inside diameter is 140 to 150 μm for oocytes/cleavage-stage embryos and 200 to 250 μm for blastocysts.
The liquid surface in particular can be identified by focusing on the edge of the dispensed reagent. If you continue to use it, you will get a sense of the distance between the top and bottom of the well. Please practice it several times.
All operations should be performed at room temperature of 25 -27°C, at which no irreversible change will be caused by temperature (temperature drop causes no damage) during operation of oocytes/embryos for 30 minutes, the time obtained by back-calculation. Therefore, all operations per oocyte/embryo should be strictly performed within 30 minutes, but this is the maximum time just to respond to temperature changes. Please be careful to perform all operations with appropriate time.
Theoretically, at -196°C, all substances on earth will lose their energy and become unable to move, so they will not get better or worse. However, at this point, there are few research reports comparing them, and it can be said that long-term observation will be required in the future.
The composition of the solution differs from that of the culture system. Increasing the temperature of the freezing/thawing solution increases the toxicity inherent in CPA. Also, since the osmotic pressure changes due to volatilization, it is desirable to keep the temperature as low as possible. However, as cells die when the temperature is lowered, operation should be performed at 25 - 27°C, the temperature which does not drop to the temperature range causing irreversible damage even after 30 minutes of operation.
The design allows for easiness to hold and an increased flat area, making it possible to affix a sticker avoiding the logo and write many letters.
Fertilization rate and culture results are affected by various factors such as previous procedures and the original quality of oocytes. It is difficult to identify the definitive cause. We recommend that you first check whether each procedure is performed according to the protocol and review other culture environments, etc. Especially for poor fertilization rates in ICSI, the total time from HCG administration to the end of ICSI (excluding the freezing period) is also important, so please check if culture time is not too long.
The tip of Cryotec is cut into an irregular triangle. When the sheet is facing up with the logo on the right, the sharp corner of the triangle will be above the view, so you can identify it.
We recommend the use of a small cooling rack that can be purchased from our company. If a large rack is used, evaporation occurs fast and the surrounding temperature is likely to be lowered due to the large surface area. Therefore, please pay more attention, such as not placing it near the microscope until immediately before use and replenish frequently so that the volume of liquid nitrogen will not be reduced.
The volume of the reagent to be dispensed is minimum 300 µL, which is enough for continuous use without the lid, so the operation can be performed without the lid. At the time of thawing, it is important to dilute slowly by mixing the previous solution and the current solution over time. Pay attention to vibration caused by opening and closing of the lid.
All products can be purchased individually.
