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Method for observing collagenous fibre of fresh and alive trepang with electron microscope

A collagen fiber and sea cucumber technology is applied in the field of electron microscope observation of sea cucumber collagen fibers in a fresh state, which can solve the problems of collagen denaturation and cannot reflect the existing state, and achieve the effect of easy deterioration.

Inactive Publication Date: 2009-09-30
OCEAN UNIV OF CHINA
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Problems solved by technology

However, regardless of the extraction method, the collagen will be denatured to varying degrees, and the extracted single collagen cannot reflect its true state of existence in muscle tissue.

Method used

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  • Method for observing collagenous fibre of fresh and alive trepang with electron microscope

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Embodiment Construction

[0008] Cut sea cucumber body wall muscle into 5×5×3mm 3 The blocks were washed with 0.1M pH 7.4 phosphate buffer, and fixed with 0.5M glutaraldehyde phosphate buffer with pH 7.2 at 4°C for 2 hours. Sea cucumber samples were soaked in 2M NaOH aqueous solution for 4 hours at 20°C, washed with 5% normal saline to pH 7.2, and then stained with 2% tannic acid. Hour. Fix with glutaraldehyde phosphate buffer solution with a concentration of 0.5M and a pH of 7.2 for 30 minutes at 4°C, and use alcohols with concentrations ranging from low to high of 50%, 60%, 70%, 80%, 90%, and 100%. (V / V) After successive dehydration, carry out critical point drying, the critical point temperature is 31.4°C and the pressure is 39.5kg / cm 2 , observed with a scanning electron microscope after gold-plated film treatment.

[0009] The concentration of the phosphate buffer described in the present invention is 0.1M-1M, and the pH is 6.0-8.0; the concentration of the glutaraldehyde phosphate buffer is 0....

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Abstract

The invention relates to a method for observing the collagenous fibre of fresh and alive trepang with an electron microscope, which is characterized by comprising the steps: firstly, the body wall muscles of the trepang are cut into pieces, the trepang pieces are cleanly washed by phosphoric acid buffer solution, the cleanly washed trepang pieces are fixed by glutaraldehyde phosphoric acid buffer solution for 0.5-5 hours under the temperature of -5-10 DEG C, and a trepang sample which is soaked in NaOH aqueous solution for 1-10 hours under the temperature of 10-50 DEG C is washed by normal saline solution until the PH value reaches 6.0-8.0; and secondly, the trepang sample is dyed by tannin, fixed by the glutaraldehyde phosphoric acid buffer solution under the temperature of 0-10 DEG C, sequentially dehydrated by alcohol with concentration ranging from low to high, dried at critical point, processed by gilded film and observed by a scanning electron microscope. Because trepang sample has the characteristics of easy putrescence, easy degenerescence of the marine product and isophagy per se, the two steps of collagenous fibre fixation and conductive dying are added to the preparation process of the electron microscope sample thereof, and the tissue constitution of the collagenous fibre of the fresh and alive trepang can be observed, thereby achieving the ideal effect.

Description

technical field [0001] The invention relates to a marine biological food sea cucumber collagen fiber, in particular to an electron microscope observation method of sea cucumber collagen fiber in a fresh state. Background technique [0002] Sea cucumber is listed as one of the "Eight Treasures of Seafood" because of its high nutritional and pharmaceutical value. It is a good product for nourishing, fitness, curing diseases and preventing aging. Sea cucumber body wall muscles account for more than 85% (M / M) of the total weight of sea cucumbers, which are composed of dermal connective tissue, and more than 70% of the main component of dermal connective tissue is collagen fibers. It can be said that the texture characteristics of sea cucumbers are to a large extent The above is determined by the tissue structure characteristics of collagen fibers. Therefore, it is of great significance to observe the tissue structure of sea cucumber collagen fibers in the fresh state and its ex...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N23/225G01N1/30
Inventor 高昕张朝辉李昭勇许加超陈燕
Owner OCEAN UNIV OF CHINA
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