Cryopreservation box in prey on male mite mites sperm production applications

by:CBFI     2020-09-29
Cryopreservation box also known as the ultra-low temperature freezer, commonly used in laboratory specimens of cryopreserved production and samples and its application in prey on male mite mites sperm production:
( 1) Mating male and female mite get: from predator mites breeding populations, pick the predator mites after mite eggs kept separately into male and female mite matching, matching 10 minutes male and female mites can mate, thereby gaining mating male and female mite;

( 2) Mating male and female mites in stereotypes: 0 - after mating predator mites 120 minutes, will is in the mating process together with the feeding device in place - male and female individuals 80 ℃ or 3 - in the liquid nitrogen cryopreservation box 5 minutes, male and female mite styling;

( 3) Finalize the design after the separation of male and female mites: finalize the design after remove both male and female mites, together with the feeding device, with a fine brush pen from feeding device of mating male and female mites carry on into the glass slide, reuse 0 insects needle and fine brush tools such as separate mating male and female mite gently, to prevent the destruction of spermatophore;

( 4) Fine package won: male and female mites successfully separated, use knife cut male mite hubei body down, keep the jaw body, on the jaw body easily observed in guide essence toe at the top of the bag, then fine package with a fuzz tip gently pick down into 70% Save or use directly in 100% alcohol;

( 5) Sperm extract and glass samples preparation: use glue dropper head drops into a drop of 0. 075 mol/L of potassium chloride solution in the middle of the slide, and then use 0 fine brush to prey on mite spermatophore placed on the slide of potassium chloride solution in room temperature for 30 minutes, reoccupy pipetting gun head or glue dropper will suck or potassium chloride solution will be fine for writing brush is transferred to the new slide, with 0 insects needle or a finer gently broken needle tool will fine package, glass slide and quickly to drip a drop of the new configuration of stationary liquid ( Methanol, acetic acid glacial = 3:1) , fixed 30 minutes, in the process, at least three fixed complement and liquid ( Because the fixed liquid volatilization speed) , after being fixed liquid, dry, drip into the dye solution (Jim on the slide 1 concentrate, PH = 7. 9 diluted phosphoric acid buffer of 2) , under the normal temperature dyeing 60 minutes, with clear water to rinse the dye solution, finally after natural drying or use the hair dryer blow dry, the location of the dyes on the slide was above drip a drop of optimal barak, then cover the cover glass, and the circle marker dyes on the part of the note for easy observation, after being fixed sample can be observed under optical microscope predator mites mite male sperm.



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