CN112857539A - Method for weighing zebra fish embryos and juvenile fish - Google Patents
Method for weighing zebra fish embryos and juvenile fish Download PDFInfo
- Publication number
- CN112857539A CN112857539A CN202110023911.6A CN202110023911A CN112857539A CN 112857539 A CN112857539 A CN 112857539A CN 202110023911 A CN202110023911 A CN 202110023911A CN 112857539 A CN112857539 A CN 112857539A
- Authority
- CN
- China
- Prior art keywords
- filter membrane
- weighing
- suction filtration
- funnel
- weight
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01G—WEIGHING
- G01G17/00—Apparatus for or methods of weighing material of special form or property
Abstract
The invention discloses a method for weighing zebra fish embryos and juvenile fish. After a sample to be weighed is washed by fresh culture solution, the zebra fish embryo or juvenile fish is sucked and put into a centrifuge tube for standby; putting the filter membrane into a vacuum filtration funnel, adding deionized water, starting a vacuum pump, and timing and performing filtration after no visible liquid exists above the funnel; after the suction filtration is finished, taking down the filter membrane, placing the filter membrane in a dryer, and weighing and recording the weight as M1; repeating the steps, and weighing and recording the weight as M2; putting the cellulose acetate filter membrane into a suction filtration device, adding deionized water, starting a vacuum pump, dripping N zebra fish embryos onto the filter membrane after no visible liquid exists above a funnel, and performing suction filtration at regular time; taking down the filter membrane, and weighing the filter membrane as M3; putting the filter membrane into a suction filtration device, adding deionized water, starting a vacuum pump, timing and carrying out suction filtration after no visible liquid exists above the funnel, taking down the filter membrane, and weighing and recording the weight as M4; and calculating the weight M of the zebra fish embryo, namely (M4+ M3-M2-M1)/2N. The method has accurate weighing.
Description
Technical Field
The invention relates to the technical field of life science, in particular to a method for quantifying the weight of zebra fish embryos, which is suitable for quantifying the weight of the zebra fish embryos and young fishes.
Background
Zebrafish are highly homologous to human genes, which means that experimental results for zebrafish are mostly applicable to humans, and thus zebrafish is a very widely used model organism. The zebra fish is mild in sexual condition and strong in fertility, the mature zebra fish can lay eggs once every few days, 200 eggs can be laid each time, and embryos can be developed and formed within 24 hours, so that experimenters can perform different experiments on the same generation of fish, and further study the pathological evolution process and find out the causes of diseases. In addition, the zebra fish embryo is transparent in whole body, each tissue under a microscope is clear and visible, and experimenters can easily observe the influence of the medicine or the poison on the internal organs of the zebra fish embryo, so that the zebra fish embryo is widely used for medicine research and toxicology research. The weight of zebra fish embryos is also an important index in toxicological studies, however, there is no method for simply and accurately quantifying the weight of zebra fish embryos.
Disclosure of Invention
In view of the problems in the prior art, the invention provides a method for weighing zebra fish embryos and juvenile fish.
The invention aims to solve the technical problems, adopts the technical scheme that the method for weighing the weight of the zebra fish embryo and the weight of the juvenile fish comprises the following steps:
(1) after a sample to be weighed is washed by fresh culture solution, the zebra fish embryo or juvenile fish is sucked and put into a centrifuge tube for standby;
(2) putting the filter membrane into a vacuum filtration funnel, adding deionized water, starting a vacuum pump, and timing and performing filtration after no visible liquid exists above the funnel;
(3) after the suction filtration is finished, taking down the filter membrane, placing the filter membrane in a dryer, and weighing and recording the weight as M1;
(4) repeating the steps (2) and (3), and weighing and recording the weight as M2;
(5) putting the cellulose acetate filter membrane in the step (4) into a suction filtration device, adding deionized water, starting a vacuum pump, sucking N zebra fish embryos from the centrifugal tube in the step (1) by using a suction tube after no visible liquid exists above the funnel, dripping the N zebra fish embryos onto the filter membrane, and performing suction filtration at regular time;
(6) taking down the filter membrane, and weighing the filter membrane as M3;
(7) putting the filter membrane in the step (6) into a suction filtration device, adding deionized water, starting a vacuum pump, timing suction filtration after no visible liquid exists above the funnel, taking down the filter membrane, and weighing and recording the weight as M4;
(8) and calculating the weight M of the zebra fish embryo, namely (M4+ M3-M2-M1)/2N.
The filter membrane of the invention is a cellulose acetate filter membrane with a pore size of 0.22 μm or 0.45 μm.
The diameter of the filter membrane is smaller than the inner diameter of the suction filter funnel.
Deionized water is added in the suction filtration process, and the volume of the deionized water is 50ml-100 ml.
The suction filtration time of the invention is 20min-40 min.
The weighing tool is a one-tenth-of-a-thousand balance or a one-tenth-of-a-ten-thousandth balance.
The fresh culture solution is E3 culture solution, and its components are 5mmol/L NaCl, 0.17mmol/L KCl, 0.33mmol/L CaCl2，0.33mmol/L MgSO4，pH 7.4。
Has the advantages that:
the method is simple to operate, uses conventional laboratory instruments, is simple and convenient in calculation method, and does not cause burden on subsequent experiments of researchers. Can accurately weigh the weight of single or multiple zebra fish embryos or juvenile fish.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the accompanying tables.
The fresh culture solution is E3 culture solution, and its components are 5mmol/L NaCl, 0.17mmol/L KCl, and 0.33mmol/L CaCl2，0.33mmol/L MgSO4，pH 7.4。
Example 1
(1) After a sample to be weighed is washed by fresh culture solution, a zebra fish embryo or juvenile fish is sucked by a plastic suction tube and put into a 1.5ml centrifuge tube for standby;
(2) placing a cellulose acetate filter membrane with the aperture of 0.45 mu m into a vacuum filtration funnel, adding 100ml of deionized water, starting a vacuum pump, and performing vacuum filtration for 30min at a timing after no visible liquid exists above the funnel;
(3) after the suction filtration is finished, taking down the filter membrane, placing the filter membrane in a dryer, then placing the filter membrane in a one-ten-thousandth balance, and weighing and recording the weight as M1;
(4) repeating the steps (2) and (3), and weighing and recording the weight as M2;
(5) putting the cellulose acetate filter membrane in the step (4) into a suction filtration device, adding 100ml of deionized water, starting a vacuum pump, sucking 10 zebra fish embryos from a 1.5ml centrifugal tube in the step (1) by using a suction tube and dripping the zebra fish embryos onto the filter membrane after no visible liquid exists above a funnel, and performing suction filtration for 30min at regular time;
(6) taking down the filter membrane, putting the filter membrane into a one-ten-thousandth balance, and weighing the filter membrane as M3;
(7) putting the filter membrane in the step (6) into a suction filtration device, adding 100ml of deionized water, starting a vacuum pump, timing and carrying out suction filtration for 30min after no visible liquid exists above the funnel, taking down the filter membrane, putting into a one-ten-thousandth balance, and weighing the filter membrane as M4;
(8) zebrafish embryo weight M ═ (M4+ M3-M2-M1)/(2 x 10).
As can be seen from Table 1, the weight of a single zebra fish embryo is measured to be 0.057mg, the standard deviation is 0.008, the coefficient of variation is 13.478%, and the repeatability is good.
TABLE 1 EXAMPLE 1 Zebra fish embryo weight (in mg) was measured in triplicate
Example 2
(1) After a sample to be weighed is washed by fresh culture solution, a zebra fish embryo or juvenile fish is sucked by a plastic suction tube and put into a 1.5ml centrifuge tube for standby;
(2) placing a cellulose acetate filter membrane with the aperture of 0.45 mu m into a vacuum filtration funnel, adding 100ml of deionized water, starting a vacuum pump, and performing vacuum filtration for 30min at a timing after no visible liquid exists above the funnel;
(3) after the suction filtration is finished, taking down the filter membrane, placing the filter membrane in a dryer, then placing the filter membrane in a one-ten-thousandth balance, and weighing and recording the weight as M1;
(4) repeating the steps (2) and (3), and weighing and recording the weight as M2;
(5) putting the cellulose acetate filter membrane in the step (4) into a suction filtration device, adding 100ml of deionized water, starting a vacuum pump, sucking 30 zebra fish embryos from the 1.5ml centrifugal tube in the step (1) by using a suction tube and dripping the zebra fish embryos onto the filter membrane after no visible liquid exists above the funnel, and performing suction filtration for 30min at regular time;
(6) taking down the filter membrane, putting the filter membrane into a one-ten-thousandth balance, and weighing the filter membrane as M3;
(7) putting the filter membrane in the step (6) into a suction filtration device, adding 100ml of deionized water, starting a vacuum pump, timing and carrying out suction filtration for 30min after no visible liquid exists above the funnel, taking down the filter membrane, putting into a one-ten-thousandth balance, and weighing the filter membrane as M4;
(8) zebrafish embryo weight M ═ (M4+ M3-M2-M1)/(2 x 30).
As can be seen from Table 2, the weight of a single zebra fish embryo is measured to be 0.054mg, the standard deviation is 0.007, the coefficient of variation is 12.499%, and the repeatability is good.
Table 2 example 2 repeated measurements of zebrafish embryo weight (in mg) three times
Example 3
(1) After a sample to be weighed is washed by fresh culture solution, a zebra fish embryo or juvenile fish is sucked by a plastic suction tube and put into a 1.5ml centrifuge tube for standby;
(2) placing a cellulose acetate filter membrane with the aperture of 0.45 mu m into a vacuum filtration funnel, adding 100ml of deionized water, starting a vacuum pump, and performing vacuum filtration for 30min at a timing after no visible liquid exists above the funnel;
(3) after the suction filtration is finished, taking down the filter membrane, placing the filter membrane in a dryer, then placing the filter membrane in a one-ten-thousandth balance, and weighing and recording the weight as M1;
(4) repeating the steps (2) and (3), and weighing and recording the weight as M2;
(5) putting the cellulose acetate filter membrane in the step (4) into a suction filtration device, adding 100ml of deionized water, starting a vacuum pump, sucking 50 zebra fish embryos from a 1.5ml centrifugal tube in the step (1) by using a suction tube after no visible liquid exists above a funnel, dropwise adding the zebra fish embryos onto the filter membrane, and performing suction filtration for 30min at regular time;
(6) taking down the filter membrane, putting the filter membrane into a one-ten-thousandth balance, and weighing the filter membrane as M3;
(7) putting the filter membrane in the step (6) into a suction filtration device, adding 100ml of deionized water, starting a vacuum pump, timing and carrying out suction filtration for 30min after no visible liquid exists above the funnel, taking down the filter membrane, putting into a one-ten-thousandth balance, and weighing the filter membrane as M4;
(8) zebrafish embryo weight M ═ (M4+ M3-M2-M1)/(2 x 50).
As can be seen from Table 3, the weight of the individual zebrafish embryos measured was 0.05mg, the standard deviation was 0.003, the coefficient of variation was 6.387%, and the reproducibility was very good.
Table 3 example 3 repeated measurements of zebrafish embryo weight (in mg) three times
Example 4
(1) After a sample to be weighed is washed by fresh culture solution, a zebra fish embryo or juvenile fish is sucked by a plastic suction tube and put into a 1.5ml centrifuge tube for standby;
(2) placing a cellulose acetate filter membrane with the aperture of 0.45 mu m into a vacuum filtration funnel, adding 100ml of deionized water, starting a vacuum pump, and performing vacuum filtration for 30min at a timing after no visible liquid exists above the funnel;
(3) after the suction filtration is finished, taking down the filter membrane, placing the filter membrane in a dryer, then placing the filter membrane in a one-ten-thousandth balance, and weighing and recording the weight as M1;
(4) repeating the steps (2) and (3), and weighing and recording the weight as M2;
(5) putting the cellulose acetate filter membrane in the step (4) into a suction filtration device, adding 100ml of deionized water, starting a vacuum pump, sucking 70 zebra fish embryos from a 1.5ml centrifugal tube in the step (1) by using a suction tube and dripping the zebra fish embryos onto the filter membrane after no visible liquid exists above a funnel, and performing suction filtration for 30min at regular time;
(6) taking down the filter membrane, putting the filter membrane into a one-ten-thousandth balance, and weighing the filter membrane as M3;
(7) putting the filter membrane in the step (6) into a suction filtration device, adding 100ml of deionized water, starting a vacuum pump, timing and carrying out suction filtration for 30min after no visible liquid exists above the funnel, taking down the filter membrane, putting into a one-ten-thousandth balance, and weighing the filter membrane as M4;
(8) zebrafish embryo weight M ═ (M4+ M3-M2-M1)/(2 × 70).
As can be seen from Table 4, the weight of the individual zebrafish embryos measured was 0.057mg, the standard deviation was 0.001, the coefficient of variation was 1.845%, and the reproducibility was very good.
Table 4 example 4 repeated measurements of zebrafish embryo weight (in mg) three times
Example 5
(1) After a sample to be weighed is washed by fresh culture solution, a zebra fish embryo or juvenile fish is sucked by a plastic suction tube and put into a 1.5ml centrifuge tube for standby;
(2) placing a cellulose acetate filter membrane with the aperture of 0.45 mu m into a vacuum filtration funnel, adding 100ml of deionized water, starting a vacuum pump, and performing vacuum filtration for 30min at a timing after no visible liquid exists above the funnel;
(3) after the suction filtration is finished, taking down the filter membrane, placing the filter membrane in a dryer, then placing the filter membrane in a one-ten-thousandth balance, and weighing and recording the weight as M1;
(4) repeating the steps (2) and (3), and weighing and recording the weight as M2;
(5) putting the cellulose acetate filter membrane in the step (4) into a suction filtration device, adding 100ml of deionized water, starting a vacuum pump, sucking 90 zebra fish embryos from the 1.5ml centrifugal tube in the step (1) by using a suction tube and dripping the zebra fish embryos onto the filter membrane after no visible liquid exists above the funnel, and performing suction filtration for 30min at regular time;
(6) taking down the filter membrane, putting the filter membrane into a one-ten-thousandth balance, and weighing the filter membrane as M3;
(7) putting the filter membrane in the step (6) into a suction filtration device, adding 100ml of deionized water, starting a vacuum pump, timing and carrying out suction filtration for 30min after no visible liquid exists above the funnel, taking down the filter membrane, putting into a one-ten-thousandth balance, and weighing the filter membrane as M4;
(8) zebrafish embryo weight M ═ (M4+ M3-M2-M1)/(2 x 90).
As can be seen from Table 5, the weight of a single zebra fish embryo measured was 0.053mg, the standard deviation was 0.002, the coefficient of variation was 3.193%, and the reproducibility was very good.
TABLE 5 EXAMPLE 5 repeated three measurements of zebrafish embryo weight (in mg)
As an example, a cellulose acetate filter with a pore size of 0.22 μm can be used, the volume of deionized water can be 100ml, the suction filtration time ranges from 20min to 40min, and the weighing tool is a ten-thousandth balance.
The difference between the above-mentioned embodiments 1, 2, 3, 4 and 5 is that the number of zebrafish embryos measured at the same time is different, and from the measurement results, the reproducibility is good no matter how many embryos are measured at the same time, and the reproducibility becomes good as the number of embryos increases.
Claims (7)
1. A method for weighing zebra fish embryos and juvenile fish is characterized by comprising the following steps:
(1) after a sample to be weighed is washed by fresh culture solution, the zebra fish embryo or juvenile fish is sucked and put into a centrifuge tube for standby;
(2) putting the filter membrane into a vacuum filtration funnel, adding deionized water, starting a vacuum pump, and timing and performing filtration after no visible liquid exists above the funnel;
(3) after the suction filtration is finished, taking down the filter membrane, placing the filter membrane in a dryer, and weighing and recording the weight as M1;
(4) repeating the steps (2) and (3), and weighing and recording the weight as M2;
(5) putting the cellulose acetate filter membrane in the step (4) into a suction filtration device, adding deionized water, starting a vacuum pump, sucking N zebra fish embryos from the centrifugal tube in the step (1) by using a suction tube after no visible liquid exists above the funnel, dripping the N zebra fish embryos onto the filter membrane, and performing suction filtration at regular time;
(6) taking down the filter membrane, and weighing the filter membrane as M3;
(7) putting the filter membrane in the step (6) into a suction filtration device, adding deionized water, starting a vacuum pump, timing suction filtration after no visible liquid exists above the funnel, taking down the filter membrane, and weighing and recording the weight as M4;
(8) and calculating the weight M of the zebra fish embryo, namely (M4+ M3-M2-M1)/2N.
2. The method of claim 1, wherein the filter is a cellulose acetate filter with a pore size of 0.22 μm or 0.45 μm.
3. The method of claim 1, wherein the diameter of the filter membrane is less than the inner diameter of the suction funnel.
4. The method of claim 1, wherein deionized water is added during the suction filtration process in a volume of 50ml to 100 ml.
5. The method according to claim 1, wherein the suction filtration time is 20min to 40 min.
6. The method of claim 1, wherein the weighing instrument is a one-hundred-thousandth balance or a one-ten-thousandth balance.
7. The method of claim 1, wherein the fresh culture medium is E3 culture medium comprising 5mmol/L NaCl, 0.17mmol/L KCl, and 0.33mmol/L CaCl2，0.33mmol/L MgSO4，pH 7.4。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110023911.6A CN112857539B (en) | 2021-01-08 | 2021-01-08 | Method for weighing zebra fish embryos and juvenile fish |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110023911.6A CN112857539B (en) | 2021-01-08 | 2021-01-08 | Method for weighing zebra fish embryos and juvenile fish |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112857539A true CN112857539A (en) | 2021-05-28 |
| CN112857539B CN112857539B (en) | 2022-07-15 |
Family
ID=76005503
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110023911.6A Active CN112857539B (en) | 2021-01-08 | 2021-01-08 | Method for weighing zebra fish embryos and juvenile fish |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112857539B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113447104A (en) * | 2021-06-25 | 2021-09-28 | 成都理工大学 | Method for measuring fresh weight of miniature aquatic organisms, miniature device and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102899385A (en) * | 2012-11-13 | 2013-01-30 | 中国医学科学院药用植物研究所 | Application of zebra fish to testing safety of safe substance and method for applying zebra fish to test safety of safe substance |
| CN111678833A (en) * | 2020-06-03 | 2020-09-18 | 唐山三友化工股份有限公司 | Method for measuring content of water insoluble substances in soda ash by using microporous filter membrane |
-
2021
- 2021-01-08 CN CN202110023911.6A patent/CN112857539B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102899385A (en) * | 2012-11-13 | 2013-01-30 | 中国医学科学院药用植物研究所 | Application of zebra fish to testing safety of safe substance and method for applying zebra fish to test safety of safe substance |
| CN111678833A (en) * | 2020-06-03 | 2020-09-18 | 唐山三友化工股份有限公司 | Method for measuring content of water insoluble substances in soda ash by using microporous filter membrane |
Non-Patent Citations (1)
| Title |
|---|
| 方韬 等: "水中悬浮物测定方法探讨", 《能源环境保护》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113447104A (en) * | 2021-06-25 | 2021-09-28 | 成都理工大学 | Method for measuring fresh weight of miniature aquatic organisms, miniature device and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112857539B (en) | 2022-07-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112857539B (en) | Method for weighing zebra fish embryos and juvenile fish | |
| Stalker et al. | Seasonal variation in the morphology of Drosophila robusta Sturtevant | |
| CN109220902A (en) | A kind of producing method for seed of hydrostatic pressing induction rainbow trout triploid | |
| SE534887C2 (en) | Procedure for stratification and storage of somatic embryos | |
| CN102577852A (en) | Method for identifying salt tolerance of cotton | |
| CN103609439A (en) | Influence and application method of different strains on hairy roots of ginseng | |
| CN113179898A (en) | Resistance identification method for improving infection efficiency of soybean cyst nematodes | |
| CN110794024A (en) | Standardized determination method for shellfish blood physiological indexes | |
| Kramer et al. | The nature of the auxin in tomato stem tips | |
| CN103070141B (en) | Method for testing development process of silkworm embryos | |
| Botti et al. | Inflorescence bud induction in Vitis vinifera L. cv. Thompson Seedless: Cytohistological events and starch accumulation in the shoot apex | |
| Cheng | On the fertility of marine Cladocera with a note on the formation of the resting egg in Evadne nordmanni Lovén and Podon intermedius Lilljeborg | |
| Mueller | Studies on Cultural Requirements of Bacteria: IV. Quantitative Estimation of Bacterial Growth | |
| Birky Jr et al. | Nuclear number in the rotifer Asplanchna: intraclonal variation and environmental control | |
| CN102586392B (en) | Quality detection method for unfertilized egg of turbo | |
| CN102998198A (en) | Device and method for measuring specific growth rate of plant in-vitro root | |
| CN102524168B (en) | Comprehensive assessment selection method of disease resistance of egg-laying ducks | |
| CN102177874B (en) | Method for detecting embryonic development process of bombyx mori L. eggs | |
| Horwitz | Rates of oxygen consumption of fertilized and unfertilized Asterias, Arbacia, and Spisula eggs | |
| Hansen et al. | Rate of development, viability, vigor, and virulence of Ascaridia galli ova cultured respectively in air and in water | |
| Joseph | Refractometry of fungi | |
| CN114847074B (en) | Evaluation method for cold resistance of rubber tree | |
| CN113670875B (en) | Method for screening antioxidant by using delicious euglena | |
| Pearl | On the distribution of differences in vitality among individuals | |
| CN106171133B (en) | A kind of method for culturing seedlings of radices trichosanthis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |