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A Histological Method to Distinguish Three Kinds of Muscle Cells in Carp

A technique of muscle cells and histology, which is applied in the field of microscopic observation of samples, can solve the problems of long preparation time and achieve the effect of easy observation and maintaining integrity

Active Publication Date: 2020-11-10
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, during the preparation of HE stained samples, the requirements for the producer are relatively strict in terms of staining time, tissue block size, temperature, etc., and the preparation time is long.

Method used

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  • A Histological Method to Distinguish Three Kinds of Muscle Cells in Carp
  • A Histological Method to Distinguish Three Kinds of Muscle Cells in Carp
  • A Histological Method to Distinguish Three Kinds of Muscle Cells in Carp

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A histological method for distinguishing three kinds of carp muscle cells, including freezing, freezing, pretreatment, ATPase reaction, color development, and optical microscope observation, the specific steps are:

[0021]1) Freezing: Wash the sample block taken from the back of the carp with a cryoprotectant with a concentration of 2.1% for 12 minutes, put it on the sample stage, add the frozen section embedding agent dropwise, and then put the sample and the sample stage into a container of isoamyl In the container of alkane, put the container into liquid nitrogen and freeze it to the freezing point quickly, the above-mentioned cryoprotectant is trehalose and glycoside, and its weight ratio is 1:0.62, and the ratio of levoside and dextran in glycoside is 1:90, during the freeze-drying process, a reasonable ratio of levoside and dextran can increase the viscosity of the solution, making the glucoside produce steric hindrance between protein molecules in muscle cells, a...

Embodiment 2

[0030] A histological method for distinguishing three kinds of carp muscle cells, the specific steps are:

[0031] 1) Freezing: Wash the sample block taken from the back of the carp with a cryoprotectant with a concentration of 3% for 9 minutes, put it on the sample stage, add the frozen section embedding agent dropwise, and then put the sample and the sample stage into a container of isoprene. In the container of alkane, put the container into liquid nitrogen and quickly freeze to the freezing point. The above-mentioned cryoprotectant is trehalose and glycoside, and its weight ratio is 1:0.8. The ratio of levoside and dextran in glycoside is 1:84;

[0032] 2) Slicing: Put the sample that reached the freezing point into a cryostat at -28°C for 55 minutes, then fix the sample stage to a cryostat and cut out a slice with a thickness of 11 µm, collect the cut sample on a glass slide, Dry at room temperature to obtain sample slices;

[0033] 3) Pretreatment: Fill the pretreatmen...

Embodiment 3

[0038] A histological method for distinguishing three kinds of carp muscle cells, the specific steps are:

[0039] 1) Cut a 1×1×1cm sample block from the back pectoral fin of carp, wash it with a cryoprotectant with a concentration of 2.5% for 10 minutes, put it on the sample stage, add the frozen section embedding agent dropwise, and place the sample and the sample stage. Put into the container that isopentane is housed, and put the container into liquid nitrogen and quickly freeze it to the freezing point. The ratio of glycosides is 1:88;

[0040] 2) Put the sample that reached the freezing point into a cryostat at -25°C for 60 minutes, then fix the sample stage to a cryostat and cut out a slice with a thickness of 10 µm, collect the cut sample on a glass slide, and store it at room temperature Dried to obtain sample slices;

[0041] 3) Fill the pretreatment container with 0.1M tryprotinin buffer and 18mM CaCl 2 The pH of the pretreatment solution is 10.60, and the pretre...

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Abstract

The invention discloses a histological method for distinguishing three kinds of carp muscle cells, wherein the method comprises the specific steps: freezing a treated sample with isoamyl and liquid nitrogen; freezing the sample at low temperature, slicing, and drying at room temperature; putting the slice into a constant-temperature pretreatment liquid, treating, and then cleaning with a cleaningliquid; adding a cleaning liquid into the slice, pouring out after the temperature is constant, adding an ATPase reaction liquid with the same temperature, carrying out a reaction, and cleaning and impregnating with distilled water; soaking the slice with a CaCl2 solution, a CoCl2 solution and an ammonium sulfide solution successively, and cleaning and impregnating with distilled water after the soaking is finished; absorbing excess moisture of the slice, covering with cover glass, sealing the slice, observing with an optical microscope, and thus distinguishing the three kinds of carp muscle cells. Boundary lines of red meat cells, pink meat cells and white meat cells in microscope images obtained by the distinguishing method are obvious, and the distribution of the three kinds of carp muscle cells can be clearly distinguished.

Description

technical field [0001] The invention relates to the technical field of microscopic observation samples, in particular to a histological method for distinguishing three kinds of carp muscle cells. Background technique [0002] Carp is one of the most widely distributed and most productive freshwater fish. Because carp has low fat content and rich protein content, it has become one of the most popular freshwater fish. The smallest structural unit in carp muscle is muscle fiber, and each muscle fiber is a cell. Compared with other animals, the muscle fibers of fish are shorter and thinner, the protein tissue is loose, the water content is high, and the muscle fibers are denser. These structural characteristics make fish meat softer than the muscles of other animals, so it is often used as animal protein. Resources for making food ingredients. However, based on the above characteristics, carp meat is also more likely to be degraded, which softens carp muscle during low-tempera...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/00G01N1/42G01N1/30G01N1/06
CPCG01N1/06G01N1/30G01N1/42G01N21/00
Inventor 梁佳
Owner ZHEJIANG OCEAN UNIV
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