Live Tumor Tissue Cryopreservation Kit (LT2601)
Tissue cryotube X3
Tissue support X3
Tissue cryopreservation:
No.1 Vitrified liquid 1 (V1) ............. 1X10ml tube;
No.2 Vitrified liquid 2 (V2) ............ .....1X10ml tube

Organizational cryopreservation process:

Cleaning, processing tissue → vitrification liquid 1, 2 → liquid nitrogen vitrification → storage

Bring your own materials:

1. Saline/1XPBS 2. Ophthalmic scissors 3. Ophthalmology é•Š 4. Gingival é•Š 5. Flat é•Š 6. Tissue processing mold and matching blade (LT2603) 7. Sterile liquid nitrogen 8. Sterile liquid nitrogen box 9. Stopwatch Or timer 10. 100mm Petri dish X4 11. Sterile gauze 12. Water bath

Steps:

a. Record information about the tumor tissue on the tissue cryotube;

b. Heating in a water bath and maintaining vitrification solution 1 and vitrification solution 2 at 26 ° C; Note: When using vitrification solution 1 and vitrification solution 2 subsequently, maintaining liquid temperature at 26 ° C is beneficial to improve survival after tissue resuscitation rate.

c. In a 100 mm culture dish, the tumor tissue was washed with physiological saline, and the blood vessels, the capsule, and the necrotic tissue were removed using an ophthalmic scissors and an ophthalmological forceps;

d. Cut the tumor tissue into 1 mm thick slices using a tissue processing mold; Note: The optimal thickness of the tissue section is verified to be 1 mm.

e. Wash the cut tissue with physiological saline again in a 100 mm culture dish, and aspirate the liquid on the surface of the tissue with sterile gauze;

f. Using a flat mouth sputum, immerse the tumor tissue section in 10 ml vitrification solution 1 in V1 tube, 26 ° C, 25 min;

g. Pour all the liquid and tissue sections from the V1 tube into a 100mm culture dish. Use sterile gauze to remove the vitrification solution 1 from the tissue surface before moving it into the V2 tube. Note: Minimize the tissue section from the vitrification solution. 1 The operating time between moving into vitrification solution 2.

h. Using a flat file, transfer the tissue section into 10 ml of vitrification solution 2 in a V2 tube, and allow the tissue section to settle naturally to the bottom of the tube at 26 ° C for 15 min. Note: If the tissue section does not naturally settle to the tube of the V2 tube within 15 min Bottom, wait until it sinks to the bottom of the tube; if the tissue section sinks to the bottom of the tube earlier than 15 min, allow the tissue section to soak in the V2 tube for at least 15 min.

i. Place the tissue slices on the tissue scaffold;

j. Place the tissue scaffold on a sterile gauze and try to absorb the liquid on the surface of the tissue section;

k. Quickly immerse the tissue scaffold into sterile liquid nitrogen using a gingival sputum for a period of not less than 5 min;

l. Quickly move the tissue scaffold into the tissue cryotube, screw the cryotube, and transfer the cryotube to the liquid nitrogen tank for storage. Note: Before the tissue scaffold is moved into the tissue cryotube, check that the tissue section is transparent. Transparent tissue sections mean that the tissue section is successfully vitrified.

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