How to Use ivitri - Straw system? Manufacture Instruction

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1 How to Use ivitri - Straw system? Manufacture Instruction

2 Information of ivitri -Straw ivitri is individual Vitrification Device, called ivitri. ivitri -Straw is US patent and patent pending, and registered product (USPTO), specifically designed for vitrification by cooperation between physics and math scientists, and top embryologist according to liquid-device dynamics (Liquid droplet radius is changed base on liquid volume, however, external osmolality is not changed with vary liquid volume), which can be effectively used for freezing oocyte, embryos and reproductive tissues (ovary and TESE) in human and mammalian IVF.

3 Production of Package

4 Product is individual wrapped and sterilized device (straw)

5 Product is individual wrapped and color sterilized device (straw)

6 Advantage of ivitri -Straw Special design material Special art and clear device Easy to use Easy to label by writing and label on the handle Color code Higher survival rate and pregnancy rate Water approval after closing cover Multiple purpose for oocyte, embryo, sperm, tissues.

7 Intended Use ivitri is consisting primarily of two parts: device body and outside cover. Each one labeled with ivitri Logo in handle as upside. Patient s name and ID or DOB can be labeled by typing and writing in this side. BBBB SSSS DOB:

8 VITRIFICATION PROCEDURE Follow up universal vitrification protocol, and takes the ivitri by the labeled side with patient name and ID or necessary information facing up.

9 Vitrification Procedure-cont. Place the ivitri under a microscope (ivitri Logo should be up) and adjust the focus on the black mark of the ivitri (face up)

10 Vitrification Procedure-cont. and put the oocytes or embryos (1~3) near to the black mark with the minimal volume of vitrification solution (1~2 μl). The black mark makes easier to put outside cover.

11 Vitrification Procedure-cont. and put the oocytes or embryos (1~3) near to the black mark with the minimal volume of vitrification solution (1.0~1.5 μl). The black mark makes easier the cover up.

12 Vitrification Procedure-cont. Rapidly plunge the ivitri into fresh LN2 holding the IVitri.

13 Vitrification Procedure-cont. Grab the cover with long tweezers (not surgical forceps) and plunge it into the liquid nitrogen until stops burbling, be aware don t take the ivitri out of the LN2 while covering up, tight and lock gently.

14 Vitrification Procedure-cont. Place the ivitri into the cane (goblet) with the cover facing down and store in LN2 tank.

15 WARMING PROCEDURE Take out the patient s canister from the Dewar and place it into a LN2 container with LN2 covering completely the ivitri, and check patient s information on the label.

16 WARMING PROCEDURE-cont. With long tweezers grabs the ivitri body and takes it off from the cane (goblet).

17 WARMING PROCEDURE-cont. With long tweezers grab the cover, and pull cover out without taking ivitri body out the LN2, leave it always inside LN2.

18 WARMING PROCEDURE-cont. Quickly take off the ivitri body (Logo face down) from LN2 and face down into pre-warmed thawing solution (drops or dish) at 37ºC on the microscope stage or laminar hold surface, and then follow the warming protocol.

19 WARMING PROCEDURE-cont. completing recovery: Oocytes for 2 hours and Embryos for 3 hours.

20 Clinical Trial Data Analysis

21 Alteration of osmolality may seriously impact on the survive rates of oocyte and embryos after warming, consequently decrease pregnancy rate

22 Liquid volume and droplet radius can directly impact on the alteration of osmolality, consequently decreased the survive rate and quality of oocyte and embryos after warming

23

24 Comparison of survive rates using three different vitridevices with loading vary volume of vitrification liquid from 0.5 to1.5µl containing bovine oocytes(900 oocytes)(%) µl 1.0µl 1.5µl cryoleaf Cryotop ivitri

25 Comparison of survive rates using three different vitridevices with loading vary volume of vitrification liquid from 0.5 to1.5µl containing oocytes and blastocyst(350 MII oocytes,360 blastocysts,2-3 oocytes or blastocysts /device)(%) Oocyte Blastocyst cryoleaf Cryotop ivitri

26 Thank you for your business!

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