Cryotech Multiple Cooling System For 100% Survival
Cryotech Vitrification is the most effective, easiest and safest method of preserving oocytes and embryos of any developmental stage. It provides very high functional survival of oocytes, assuring high rates of fertilization after ICSI and high rates of pregnancy after embryo transfer. The optimized method, “The Cryotec Method” has been developed by Dr. Masashige Kuwayama, who has introduced major advances in oocyte and embryo cryopreservation.
By strict adherence to specific details of The Cryotec Method, the clinical embryologist is assured of achieving 100% survival of normal oocytes and embryos.
Cryotech straws are CE Marked in accordance with the Medical Devices Directive 97/79/EC. Cryotech is actively working towards having accreditation for the media and plastics and expect to have this process completed during 2016. Until this time they are classed as only for research.
The Cryotec Method Perfect Survival And Safety
Cryotec Method is the most trustable method of cryopreservation.This open style method is recognized as easy, simple and repeatable for all.
Cryotec is a special device that allows to minimize the volume that will be cool and warm with a result up to 100% of survival for oocytes and all the stage of pre-implantation embryos.
CRYOTECH ADVANTAGE; “WHY 100% SURVIVAL?”
Best Vitrification Solution
- Highest Vitrification Capability by addition of HPC
- No Serum, No protein contained in solutions
- Endotoxin free Trehalose used instead of Sucrose
Best Vitrification Container; Cryotec
- Multiple cooling devise: for Closed or Open cooling
- Longer and Wider handle and sheet: easy writing and loading samples
- Safe and clear material
Best exclusive Vitri- and Warm plate
- On-focus vitrification plate
- Easy warming plate: no blind well for warming
Vitrification Protocol and Warming Protocol
The Cryotec Method is consist of 2 parts, Virification protocol and Warming protocol.
Vitrification protocol requires up to 16min for Human immature- and matured oocytes, PN zygotes, 4-8 cells embryos and blastocysts, and Warming protocol requires 10 min for all.
In The Cryotec Method, the protocol for oocytes is same to embryos one.
- Cryotech Vitrification Kit
Equilibration Solution (ES) : 1 vial of 1.0mℓ
Vitrification Solution (VS) : 2 vials of 1. 0mℓ
- Microscope (Turn off the heating plate)
- Stop watch (with count up function)
- Micro pipette for 300μℓ
Bring ES and VS vials to room temperature (25℃〜27℃) at least 1 hour before vitrification
- Fill the wells of Vitri Plate with 300µℓ of ES and 300µℓ of VS (Fig. 1 Vi).
- Put the lid on the Vitri Plate immediately.
- Observe the oocyte/embryo well and remember each perivitelline space to know the full recovery volume of then during ES.
- Aspirate the oocyte/embryo at the tip of pasteur pipette.
- Put the oocyte/embryo with a small amount of medium on the surface of the ES (Fig.2). Start counting up by the stop watch.
- Cover the Vitri Plate with the lid, and wait for the full recovery from the shrinkage.
- While waiting, open a “Cryotec package” with scissors. Write the information of the oocyte/embryo on the “handle” of Cryotec, and put it in the side groove of the Vitri Plate (Fig.3).
- Prepare fresh liquid nitrogen, and put a cover cap in it
- When the oocyte/embryo volume is completely recovered, ES equilibration ends.
If you can’t confirm the recovery completely, the limit time of ES equilibration is 15 min for the oocyte and blastocyst (160-220 µm in diameter), and 12 min for 4 – 8 cells embryo.
Figs. 4. VS1 Washing of the Oocyte/Embryo (Step 1 – 4)
- Aspirate the oocyte/embryo with a small amount of ES at the tip of the pipette･.
- Place the oocyte/embryo in mid depth of VS1 (Step 1).
- Rinse out the pipette expelling out of wells the ES left inside (Step 2/①), aspirate VS1 from the edge of the well (Step 2/②) and immediately expel it. (rinse out inside the pipette, Step 2/③)
The Oocyte/embryo will float immediately to the surface of VS1 (Step 2/④). After aspirating fresh vs from the edge, aspirate the oocyte/embryo at the tip of the pipette.
Place now the oocyte/embryo again to the bottom of VS1 (Step 3).
The oocyte/embryo will rise to mid depth and will stop (End of VS1 equilibration, Step 4).
Rinse out the pipette expelling the VS1 left inside, aspirating and expelling fresh VS2. Aspirate VS2, followed by oocyte/embryo in VS1 at the tip of the pipette.
Place the oocyte/embryo to the mid depth of the VS2 (Step 5).
Rinse out the pipette with fresh VS2 (Step 6/①②③)
Aspirate fresh VS2 from the edge of the well, mix the solution around the oocyte/embryo, and observe the complete shrinkage of the oocyte/embryo (Step 7).
Take the oocyte/embryo ａt the tip of the pipette (Step 8).
Place the oocyte/embryo near the black mark on the Cryotec sheet with small volume of the VS2. (1 oocyte/embryo per droplet is recommended, Fig. 6).
Immediately submerge and stir the Cryotec in fresh liquid nitrogen.
Put the cover cap on the Cryotec inside the liquid nitrogen.
Use the optimum size pasteur pipette.
- 140-150 μm for oocyte and cleavage stage embryo.
- 160~220 μm for blastocyst.(Depends on the size of the blastocysts)
Best timing for vitrification of blastocyst: the size(diameter) should be between 160~220 μm for perfect survival after vitirfication.
- Cryotech Warmig Kit
Warming Solution(TS) :1 vial of 1.8mℓ
Diluent Solution(DS) :1 vial of 0.5mℓ
Washing Solution(WS) :1 vial of 1.0mℓ
1 Warm Plate with 4 wells
- Microscope (Turn off the heating plate)
- Stop watch (With count up function)
- Micro pipette for 300μℓ
- Place the Warm Plate and the TS vial (with closed cap) in the incubator at 37℃ > 3 hours before warming (overnight storage is recommended).
- Bring the DS and the WS vials to room temperature (25~27℃) at least 1 hour before warming.
- Prepare fresh liquid nitrogen.
- Take a patient’s cane out of a liquid nitrogen tank, and take off the cover cap and prop up the Cryotec against inside wall the cooling rack in liquid nitrogen.
- Take the Warm Plate out of the incubator, and fill the second well with 300μℓ of the DS(Fig.7).
Figs. 8. Warming Procedure (Step 1-3)
- Take the TS vial out of the incubator, and expel all of it to the TS well (1.8 ml, Figs. 8, Step1/①)
- Quickly (within l sec) put the Cryotec from liquid nitrogen into the TS well (Figs. 8, Step1/②).
Start counting up by the stop watch for l min.
- The Oocyte/embryo releases from the Cryotec sheet by itself, and begins to float.
Figs. 9. Gradual Replacement of Solution (Dilution)
- Aspirate the oocyte/embryo first, followed by 3mm of the TS into the pipette (Figs. 9， 1).
- Introduce the TS to the bottom of the DS well (Figs. 9, 2), then expel the oocyte/embryo slowly to
- the bottom of TS layer in DS well (Figs. 9, 3), and wait for 3 min (Fig. 8, Step 2/①).
- While waiting, fill the WS1 and the WS2 well with 300μℓ each of ws Solution (Figs. 8, Step 3/①).
Figs. 10. Gradual Replacement of Solution (Washing 1 Step)
- Aspirate the oocyte/embryo followed by 3mm of the DS into the pipette (Figs. 10， 1).
- Introduce the DS to the bottom of the WS1 (Figs. 10, 2), and expel the oocyte/embryo slowly to the bottom of the DS layer in WS1 well (Figs. 10.3).
Observe the shape of the oocyte/embryo and memorize it. Turn off the light, and wait for more than 3 min.
- After 3 min, compare the shape of the oocyte/embryo to the one memorized.
Give ａ survival judgment if the shrinkage of the oocyte is recovered.
- Wait for 5 min in total (Figs. 8, Step3/②).
Washing 2 (1min)
- Aspirate the oocyte/embryo with minimal volume of the WS1 .
- Put the oocyte/embryo on the surface of the WS2 well (Fig. 8, Step3/③).
- After the oocyte/embryo sinks to the bottom, aspirate and place it on the surface of a different location, respectively.
- Put the oocyte/embryo into the droplet of the culture media until ICSI or ET.
(2 to 4 hours culture for ICSI, and 3 hours for blastocyst transfer are recommended)
Outline of Animation protocol
This animation protocol is an interactive content for you to learn the Cryotech method on your computer. In this animation protocol, you will learn the steps of Vitirification procedures and Warming procedures.
Download Animation Protocol
You can view the animation protocol on your computer without network connection only if you download the animation protocol onto your computer.
|A mount of Memory required||more than 2GB|
How to start
- Download Zip file of application.
- Decode the Zip file of application.
- Click file of “animation_protocol_130425.exe” to start application of Animation Protocol.
How to use
- The animation is played automatically. You can only view the file by clicking following menus/buttons.
- Select a step of protocol at right side menu.
- Click the “Play” button to start the animation.
- Click the “Stop” button to pause the animation.
- Hit the “Esc” key on the Upper left corner of your keyboard to close the application.
Dr. Kuwayama’s latest investigations for the best vitrification
|Vitrification||1||VITRIFICATION KIT 101||For 1 patient (3 times of Vitrification)
1 vial of ES (for 3 times vitrification)
2 vials of VS (for 3 times vitrification)
|2||VITRIFICATION SOLUTION SET 110|| For 10 times Vitrification
2 vials of ES
4 vials of VS
|Warming||3||WARMING KIT 102||For one Warming
1 vial of TS 1 vial of DS
1 vial of WS
|4||WARMING SOLUTION SET 205|| For 5 times Warming
5 vials of TS
1 vial of DS 2 vials of WS
|Cryotec Products||5||CRYOTEC VITRIFICATION
For pricing please email us on firstname.lastname@example.org
CRYOTEC VITRIFICATION CONTAINERS
Dr. Kuwayama’s best Vitrification container for human oocyte and embryo.
Multiple Cooling container: can be used as “Closed Cooling”after sealing, or “Open Cooling” before sealing.
In-focus loading of oocyte and embryo
Easy warming of vitrified oocyte and embryo
CRYOTEC VITRIFICATION SOLUTIONS
- Serum and protein free solution
- Best Vitrification capability
- With endotoxin-free Trehalose
Download the Cryotec Product Catalogue PDF