CN113355207A - Cell culture method for researching microorganisms - Google Patents

Cell culture method for researching microorganisms Download PDF

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Publication number
CN113355207A
CN113355207A CN202110123770.5A CN202110123770A CN113355207A CN 113355207 A CN113355207 A CN 113355207A CN 202110123770 A CN202110123770 A CN 202110123770A CN 113355207 A CN113355207 A CN 113355207A
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groups
cell culture
liquid outlet
wall
cell
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王晴
孙翠华
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/48—Holding appliances; Racks; Supports

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  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Clinical Laboratory Science (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The invention provides a cell culture method for microbial research, which uses a cell culture device for microbial research, wherein the cell culture device for microbial research comprises an extraction pipe, a three-way pipe, an angle valve, a liquid outlet pipe and a fixed bracket; in addition, the use of the fixed support in the invention ensures that the extraction tubes can be fixed at the mouth of the conical flask to be discharged in the process of dripping the cell sap and the chemical reagents into the conical flask for mixing, and the liquid outlet tube can be suspended at the mouth of the conical flask, thereby solving the problem that the mixing proportion error of the cell sap and the chemical reagents is large and the cell mixture is influenced to be cultured next because the cell sap and the chemical reagents are easy to remain on the inner wall of the mouth of the conical flask by manual operation in the prior art.

Description

Cell culture method for researching microorganisms
Technical Field
The invention relates to the technical field of cell culture, in particular to a cell culture method for researching microorganisms.
Background
In the research of microorganisms, cell culture is required, and the cell culture refers to a method for simulating in-vivo environment (sterility, proper temperature, pH value, certain nutritional conditions and the like) in vitro to enable the microorganisms to survive, grow and reproduce and maintain main structures and functions. Cell culture techniques are important and commonly used in cell biology research methods, and a large number of cells can be obtained through cell culture, and signal transduction, anabolism, growth and proliferation of cells and the like of the cells can be researched.
In the process of cell culture, cell sap and chemical reagents need to be mixed according to a certain proportion, however, a cell culture device in the prior art has a simple structure, cannot quantitatively extract the cell sap and the chemical reagents, and often uses a test tube to quantitatively measure the cell sap and the chemical reagents, so that operation steps are often added, and low efficiency is caused; in addition, when cell sap and chemical reagent mixing process, need to drip cell sap and chemical reagent into the erlenmeyer flask and stir the mixture, generally can adopt manual operation at the in-process of dripping into, remain cell sap and chemical reagent at the bottleneck inner wall very easily like this, lead to cell sap and chemical reagent's mixing ratio error big, influenced next to cultivate the cell mixture.
Disclosure of Invention
In order to achieve the above object, the present invention adopts the following technical scheme, a microbial research cell culture method, which uses a microbial research cell culture device, the microbial research cell culture device comprises an extraction pipe, a three-way pipe, an angle valve, a liquid outlet pipe and a fixed support, the extraction pipe is arranged at each of the left and right ends of the three-way pipe, the liquid outlet pipe is arranged at the bottom of the three-way pipe through a thread, the angle valve is arranged at the top of the three-way pipe, and the fixed support is arranged on the peripheral outer wall of the liquid outlet pipe, wherein:
the extraction pipe comprises an extraction pipe body, a piston, a thread mounting opening, a sealing cover, a push rod, liquid outlets and a needle tube, wherein the piston is arranged in an inner cavity of the extraction pipe body, the push rod is vertically arranged in the center of the top of the piston, the tail end of the top of the push rod penetrates through the surface of the top of the extraction pipe body and then is positioned right above the extraction pipe body, the liquid outlets are formed in the right side wall of the bottom of the extraction pipe body on the left side and the left side wall of the bottom of the extraction pipe body on the right side in a penetrating mode, the left end and the right end of the three-way pipe are connected with the two groups of liquid outlets through pipelines, the thread mounting opening is formed in the center of the bottom of the extraction pipe body, threads are arranged on the inner wall of the thread mounting opening, and threads matched with the thread mounting opening are formed in the peripheral outer wall of the top of the sealing cover and the peripheral outer wall of the top of the needle tube;
the fixed support comprises a fixed ring, an internal thread, positioning plates, sliding grooves, a fixed nut, an external thread, a fixed bolt, movable blocks, a positioning groove and a sponge block, the external thread is carved on the periphery of the outer wall of the surface of the liquid outlet pipe, the internal thread is carved on the periphery of the inner wall of the fixed ring, four groups of positioning plates are distributed on the periphery of the outer wall of the fixed ring in a cross shape, the sliding grooves are divided into two groups, one group of sliding grooves horizontally penetrate through the front side surface and the rear side surface of the positioning plates, the other group of sliding grooves vertically penetrate through the upper side surface and the lower side surface of the positioning plates, the two groups of sliding grooves are mutually communicated, the movable blocks are vertically and slidably arranged in the sliding grooves on the vertical surface, the top of each movable block is slidably connected with the inner cavity of the sliding groove on the horizontal surface, the surface of the sliding groove on the horizontal surface is slidably connected with the fixed bolt, and the fixed nut is installed on the end thread of the fixed bolt, a sponge block is arranged on the surface of one side of the bottom of the movable block, which faces the axis of the fixed ring, and a plurality of groups of positioning grooves distributed in a sawtooth shape are arranged on the surfaces of the front side and the rear side of the bottom of the positioning plate;
when the device is adopted to carry out microbial research cell culture, the device comprises the following steps:
s1, extracting cell sap and mixed chemical agents: the tops of the two groups of needle tubes are respectively connected with the threaded mounting ports through threads, the angle valves are closed, and then the cell sap in the glass bottle filled with the cell sap and the chemical agent in the glass bottle filled with the chemical agent are respectively and quantitatively pumped into the two groups of extraction tubes through the two groups of needle tubes according to requirements;
s2, mixing of cell sap and mixed chemical agents: after step S1, separating the needle tube from the threaded mounting port, connecting the sealing cap with the threaded mounting port by threads, then mounting the fixed bracket on the side wall of the liquid outlet tube, connecting the internal threads and the external threads by threads, placing the conical flask under the liquid outlet tube, respectively lapping the four sets of positioning plates at the mouth of the conical flask, then sliding the four sets of movable blocks until the four sets of sponge blocks are clamped with the peripheral outer wall of the mouth of the conical flask, opening the angle valve and simultaneously pushing the two sets of push rods downwards, allowing the cell sap and the chemical agent to flow into the conical flask through the two sets of liquid outlets, and finally stirring and mixing the cell sap and the chemical agent in the conical flask by the stirring rod to obtain a cell mixture;
s3, proliferation of cell culture: and (4) putting the cell mixture obtained in the step S2 into a proliferation culture medium, and performing cell culture proliferation under the conditions of sterility, non-toxicity and constant temperature.
Preferably; and the outer walls of the two groups of the extraction pipe bodies are respectively carved with a graduated scale for measuring the liquid solvent in the extraction pipe body.
Preferably; the angle valve is a two-in one-out angle valve, wherein two groups of water inlets are respectively communicated with two groups of liquid outlets, and one group of water outlet valve is communicated with the liquid outlet pipe.
Preferably; the outer diameter of the liquid outlet pipe is equal to the inner diameter of the fixing ring.
Preferably; the locating slot is made of anti-skid rubber material.
Preferably; one surface of the sponge block, which is right against the outer wall of the liquid outlet pipe, is of an arc concave surface structure.
Has the advantages that:
in addition, the use of the fixed support in the invention ensures that the extraction tubes can be fixed at the mouth of the conical flask to be discharged in the process of dripping the cell sap and the chemical reagents into the conical flask for mixing, and the liquid outlet tube can be suspended at the mouth of the conical flask, thereby solving the problem that the mixing proportion error of the cell sap and the chemical reagents is large and the cell mixture is influenced to be cultured next because the cell sap and the chemical reagents are easy to remain on the inner wall of the mouth of the conical flask by manual operation in the prior art.
Drawings
The invention is further illustrated with reference to the following figures and examples.
FIG. 1 is a flow chart of the operation of the present invention;
FIG. 2 is a schematic front view of the microbial research cell culture apparatus of the present invention in which a cell sap and a chemical agent are mixed;
FIG. 3 is a schematic front view of the cell culture apparatus for investigating microorganisms according to the present invention, in which cell sap and chemicals are extracted;
FIG. 4 is a schematic cross-sectional view of the bottom of the drain pipe of the present invention from above;
FIG. 5 is a schematic vertical sectional view of a fixing bracket according to the present invention;
FIG. 6 is an enlarged schematic view of A in FIG. 2;
fig. 7 is an enlarged structural diagram of B in fig. 2.
Detailed Description
Embodiments of the present invention will be described below with reference to the drawings. In this process, the width of the lines or the size of the components in the drawings may be exaggerated for clarity and convenience of description.
The following terms are defined based on the functions of the present invention, and may be different depending on the intention of the user or the operator or the convention. Therefore, these terms are defined based on the entire contents of the present specification.
As shown in fig. 1 to 7, a cell culture method for microbial research uses a cell culture device for microbial research, which comprises an extraction pipe 1, a three-way pipe 2, an angle valve 3, a liquid outlet pipe 4 and a fixed support 5, wherein the extraction pipe 1 is arranged at each of the left and right ends of the three-way pipe 2, the liquid outlet pipe 4 is arranged at the bottom of the three-way pipe 2 through a thread, the angle valve 3 is arranged at the top of the three-way pipe 2, and the fixed support 5 is arranged on the peripheral outer wall of the liquid outlet pipe 4;
the extraction pipe 1 comprises an extraction pipe body 11, a piston 12, a thread mounting opening 13, a sealing cover 14, a push rod 15, liquid outlets 16 and a needle tube 17, the piston 12 is arranged in an inner cavity of the extraction pipe body 11, the push rod 15 is vertically arranged in the center of the top of the piston 12, the top end of the push rod 15 penetrates through the top surface of the extraction pipe body 11 and then is positioned right above the extraction pipe body 11, the liquid outlets 16 penetrate through the right side wall of the bottom of the extraction pipe body 11 on the left side and the left side wall of the bottom of the extraction pipe body 11 on the right side, the left end and the right end of the three-way pipe 2 are connected with the two groups of the liquid outlets 16 through pipelines, the thread mounting opening 13 is drilled in the center of the bottom of the extraction pipe body 11, threads are arranged on the inner wall of the thread mounting opening 13, and threads matched with the thread mounting openings 13 are drilled on the outer walls around the top of the sealing cover 14 and the needle tube 17, the outer walls of the two groups of extraction pipe bodies 11 are respectively provided with a graduated scale for measuring a liquid solvent in the extraction pipe bodies 11, the angle valve 3 is a two-inlet one-outlet angle valve, two groups of water inlets are respectively communicated with the two groups of liquid outlets 16, and one group of water outlet valve is communicated with the liquid outlet pipe 4;
the fixed support 5 comprises a fixed ring 51, an internal thread 52, a positioning plate 53, a sliding groove 54, a fixed nut 55, an external thread 56, a fixed bolt 57, a movable block 58, a positioning groove 59 and a sponge block 510, the external thread 56 is carved on the periphery of the outer wall of the surface of the liquid outlet pipe 4, the internal thread 52 is carved on the periphery of the inner wall of the fixed ring 51, the outer diameter of the liquid outlet pipe 4 is equal to the inner diameter of the fixed ring 51, four groups of positioning plates 53 are distributed on the periphery of the outer wall of the fixed ring 51 in a cross shape, the sliding grooves 54 are divided into two groups, one group of sliding grooves 54 horizontally penetrate through the front side surface and the rear side surface of the positioning plate 53, the other group of sliding grooves 54 vertically penetrate through the upper side surface and the lower side surface of the positioning plate 53, the two groups of sliding grooves 54 are mutually communicated, the movable block 58 is vertically and slidably mounted in the sliding groove 54 on the vertical surface, and the top of the movable block 58 is slidably connected with the inner cavity of the sliding groove 54 on the horizontal plane, a fixed bolt 57 is connected to the surface of the sliding groove 54 positioned on the horizontal plane in a sliding manner, a fixed nut 55 is installed at the tail end of the fixed bolt 57 in a threaded manner, a sponge block 510 is arranged on the surface of one side of the bottom of the movable block 58, which faces the axis of the fixed ring 51, one surface of the sponge block 510, which faces the outer wall of the liquid outlet pipe 4, is of an arc concave structure, a plurality of groups of positioning grooves 59 which are distributed in a sawtooth manner are arranged on the surfaces of the front side and the rear side of the bottom of the positioning plate 53, and the positioning grooves 59 are made of an anti-skid rubber material;
when the device is adopted to carry out microbial research cell culture, the device comprises the following steps:
s1, extracting cell sap and mixed chemical agents: the tops of the two groups of needle tubes 17 are respectively connected with the threaded mounting port 13 through threads, the angle valve 3 is closed, and then the cell sap in the glass bottle filled with the cell sap and the chemical agent in the glass bottle filled with the chemical agent are respectively quantitatively extracted into the two groups of extraction tubes 1 through the two groups of needle tubes 17 according to requirements;
s2, mixing of cell sap and mixed chemical agents: after step S1, the needle tube 17 is separated from the threaded mounting port 13, the sealing cap 14 is in threaded connection with the threaded mounting port 13, then the fixing bracket 5 is mounted on the side wall of the liquid outlet tube 4, the internal thread 52 is in threaded connection with the external thread 56, the conical flask is placed under the liquid outlet tube 4, the four sets of positioning plates 53 are respectively lapped at the mouth of the conical flask, then the four sets of movable blocks 58 are slid until the four sets of sponge blocks 510 are clamped with the peripheral outer wall of the mouth of the conical flask, the angle valve 3 is opened, the two sets of push rods 15 are pushed downwards at the same time, the cell sap and the chemical agent are respectively flowed into the conical flask through the two sets of liquid outlets 16, and finally the cell sap and the chemical agent in the conical flask are stirred and mixed through the stirring rod to obtain a cell mixture;
s3, proliferation of cell culture: and (4) putting the cell mixture obtained in the step S2 into a proliferation culture medium, and performing cell culture proliferation under the conditions of sterility, non-toxicity and constant temperature.
In the process of clamping the fixed support 5 at the mouth of the conical flask, the positioning grooves 59 at the bottoms of the four groups of positioning plates 53 are respectively clamped at the top of the periphery of the mouth of the conical flask, wherein the positioning grooves 59 at the bottoms of each group of positioning plates 53 are provided with a plurality of simultaneously sliding blocks 58 which can slide along the sliding grooves 54 in the horizontal direction, so that when the fixed support 5 is required to be fixed at the mouths of the conical flasks with different calibers, the movable blocks 58 can slide towards the outer wall of the mouth while the positioning grooves 59 are clamped at the mouth again until the sponge blocks 510 are lapped on the outer wall of the mouth again, and at the moment, the fixed bolts 57 are fixed on the surface of the sliding grooves 54 by rotating the fixed nuts 55, thereby improving the universality of the invention.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (6)

1. A microbial research cell culture method uses a microbial research cell culture device, which comprises an extraction pipe (1), a three-way pipe (2), an angle valve (3), a liquid outlet pipe (4) and a fixed bracket (5), and is characterized in that: the left end and the right end of the three-way pipe (2) are respectively provided with an extraction pipe (1), the bottom of the three-way pipe (2) is provided with a liquid outlet pipe (4) through threads, the top of the three-way pipe (2) is provided with an angle valve (3), and the peripheral outer wall of the liquid outlet pipe (4) is provided with a fixed support (5);
the extraction pipe (1) comprises an extraction pipe body (11), a piston (12), a thread mounting port (13), a sealing cover (14), a push rod (15), a liquid outlet (16) and a needle tube (17), the piston (12) is arranged in an inner cavity of the extraction pipe body (11), the push rod (15) is vertically arranged in the center of the top of the piston (12), the tail end of the top of the push rod (15) penetrates through the top surface of the extraction pipe body (11) and then is positioned right above the extraction pipe body (11), the liquid outlet (16) is drilled in the right side wall of the bottom of the left extraction pipe body (11) and the left side wall of the bottom of the right extraction pipe body (11) on the right side, the left end and the right end of the three-way pipe (2) are connected with the two groups of the liquid outlets (16) through pipelines, the thread mounting port (13) is drilled in the center of the bottom of the extraction pipe body (11), the inner wall of the threaded mounting opening (13) is provided with threads, and the peripheral outer wall of the top of the sealing cover (14) and the peripheral outer wall of the top of the needle tube (17) are both provided with threads matched with the threaded mounting opening (13);
the fixed support (5) comprises a fixed ring (51), an internal thread (52), a positioning plate (53), sliding grooves (54), a fixed nut (55), an external thread (56), a fixed bolt (57), a movable block (58), a positioning groove (59) and a sponge block (510), the external thread (56) is carved on the periphery of the outer wall of the surface of the liquid outlet pipe (4), the internal thread (52) is carved on the periphery of the inner wall of the fixed ring (51), four groups of positioning plates (53) are distributed on the periphery of the outer wall of the fixed ring (51) in a cross shape, the sliding grooves (54) are divided into two groups, one group of sliding grooves (54) horizontally penetrate through the surfaces of the front side and the rear side of the positioning plate (53), the other group of sliding grooves (54) vertically penetrate through the surfaces of the upper side and the lower side of the positioning plate (53), the two groups of sliding grooves (54) are mutually communicated, and the movable block (58) is vertically and slidably mounted in the sliding grooves (54) on the vertical surface, the top of the movable block (58) is in sliding connection with an inner cavity of a sliding groove (54) located on the horizontal plane, the surface of the sliding groove (54) located on the horizontal plane is in sliding connection with a fixing bolt (57), a fixing nut (55) is installed at the tail end of the fixing bolt (57) in a threaded mode, a sponge block (510) is arranged on the surface of one side, opposite to the axis of the fixing ring (51), of the bottom of the movable block (58), and a plurality of groups of positioning grooves (59) which are distributed in a sawtooth mode are formed in the front side surface and the rear side surface of the bottom of the positioning plate (53);
when the device is adopted to carry out microbial research cell culture, the device comprises the following steps:
s1, extracting cell sap and mixed chemical agents: the tops of the two groups of needle tubes (17) are respectively connected with the thread mounting port (13) through threads, the angle valve (3) is closed, and then the cell sap in the glass bottle filled with the cell sap and the chemical agent in the glass bottle filled with the chemical agent are respectively quantitatively extracted into the two groups of extraction tubes (1) through the two groups of needle tubes (17) according to requirements;
s2, mixing of cell sap and mixed chemical agents: after step S1, separating the needle tube (17) from the thread mounting port (13), connecting the sealing cover (14) with the thread mounting port (13) in a threaded manner, then mounting the fixed support (5) on the side wall of the liquid outlet tube (4), connecting the internal thread (52) with the external thread (56) in a threaded manner, placing the conical flask under the liquid outlet tube (4), respectively lapping the four groups of positioning plates (53) at the mouth of the conical flask, then sliding the four groups of movable blocks (58) until the four groups of sponge blocks (510) are clamped with the peripheral outer wall of the mouth of the conical flask, opening the angle valve (3) and simultaneously pushing the two groups of push rods (15) downwards, allowing the cell sap and the chemical agent to flow into the conical flask through the two groups of liquid outlets (16), and finally stirring and mixing the cell sap and the chemical agent in the conical flask through the stirring rod to obtain a cell mixture;
s3, proliferation of cell culture: and (4) putting the cell mixture obtained in the step S2 into a proliferation culture medium, and performing cell culture proliferation under the conditions of sterility, non-toxicity and constant temperature.
2. The method of claim 1, wherein the cell culture medium comprises: the outer walls of the two groups of the extraction pipe bodies (11) are respectively provided with a scale for measuring the liquid solvent in the extraction pipe bodies (11).
3. The method of claim 1, wherein the cell culture medium comprises: the angle valve (3) is a two-inlet one-outlet angle valve, wherein two groups of water inlets are respectively communicated with two groups of liquid outlets (16), and one group of water outlet valve is communicated with the liquid outlet pipe (4).
4. The method of claim 1, wherein the cell culture medium comprises: the outer diameter of the liquid outlet pipe (4) is equal to the inner diameter of the fixing ring (51).
5. The method of claim 1, wherein the cell culture medium comprises: the positioning groove (59) is made of an anti-slip rubber material.
6. The method of claim 1, wherein the cell culture medium comprises: one surface of the sponge block (510) which is right against the outer wall of the liquid outlet pipe (4) is of an arc concave structure.
CN202110123770.5A 2021-01-29 2021-01-29 Cell culture method for researching microorganisms Withdrawn CN113355207A (en)

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CN202110123770.5A CN113355207A (en) 2021-01-29 2021-01-29 Cell culture method for researching microorganisms

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Application Number Priority Date Filing Date Title
CN202110123770.5A CN113355207A (en) 2021-01-29 2021-01-29 Cell culture method for researching microorganisms

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113897278A (en) * 2021-09-26 2022-01-07 李同辉 Nutrient solution adding equipment used in biological cultivation

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113897278A (en) * 2021-09-26 2022-01-07 李同辉 Nutrient solution adding equipment used in biological cultivation

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Application publication date: 20210907