Solution and ultrasonic method for extracting earthworm coelomocytes

A cell liquid and cell technology, applied in biochemical equipment and methods, microorganisms, tissue culture, etc., can solve the problems of cumbersome operation, activity damage, and low survival rate, and achieve the effect of wide practicability, easy mastery, and simple operation
CN101638634BInactive Publication Date: 2011-07-20NANJING UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
NANJING UNIV
Publication Date
2011-07-20
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a solution and an ultrasonic method for extracting earthworm coelomocytes, belonging to the technical field of bioengineering. Each 3 mL of the solution for extracting the earthworm coelomocytes contains 60-70 mM of NaCl, 5-6% by weight of ethanol, 50-60 mM of guaicol glyceryl ether and 5-6mM of EGTA PBS buffer solution. The operation steps of the ultrasonic method are as follows: step 1, clearing the bowel of an earthworm, namely, using the PBS buffer solution for repeated cleaning; step 2, putting the earthworm into the cold solution for extracting the coelomocytes, and carrying out fast ultrasonic extraction; step 3, removing the earthworm, adding LBBS buffer solution for cleaning, centrifuging and removing supernate, wherein obtained precipitates are the coelomocytes; step 4, adding cell sap for incubation; and step 5, using trypan blue for coloring and microscopic examination, and calculating cell viability. The solution and the ultrasonic method extract a great many coelomocytes, realizes high cell viability, can satisfy the needs of apoptosis and a single cell gel electrophoresis experiment, and can be used for early diagnosis of soil pollution and function for early warning.
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Description

technical field

[0001] The invention relates to an extraction solution and an ultrasonic extraction method for bioengineering technology, in particular to a solution and an ultrasonic extraction method for extracting earthworm body coelomocytes. Background technique

[0002] As an ancient organism, earthworms have existed in nature for 600 million years. They are one of the animal groups with the largest biomass in soil and are an important part of the soil system. Earthworms play important roles in the physical, chemical and biological processes of soil. They are widely distributed in the soil and play an important role in the degradation of organic matter, nutrient cycling and soil formation. At the same time, earthworms can accelerate the decomposition of carbon, affect the carbon allocation of the ecosystem, and directly or indirectly control the composition of plant communities and promote plant growth. Community succession, affecting terrestrial ecosystems. In the fo...

Claims

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