CN102533930A - Manufacturing method of slide for observing filamentous fungus microscopic morphology - Google Patents
Manufacturing method of slide for observing filamentous fungus microscopic morphology Download PDFInfo
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- CN102533930A CN102533930A CN2012100019812A CN201210001981A CN102533930A CN 102533930 A CN102533930 A CN 102533930A CN 2012100019812 A CN2012100019812 A CN 2012100019812A CN 201210001981 A CN201210001981 A CN 201210001981A CN 102533930 A CN102533930 A CN 102533930A
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- filamentous fungus
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Abstract
The invention relates to a manufacturing method of a slide for observing filamentous fungus microscopic morphology. The method comprises the following steps of: firstly digging a groove on a water agar culture flat plate and removing a culture medium in the groove; then placing a water agar culture block with a size of less than that of the groove in the groove, picking up a fungus culture to be identified and inoculating on the upward side of the water agar culture block in the groove and then covering the water agar culture block with a sterile cover glass; and closing a culture dish cover, sealing and culturing at room temperature, taking the cover glass out after the culture is finished and manufacturing an observation slide for fungus. The method provided by the invention is easy in operation for manufacturing a thallus observation slide of filamentous fungus; observation slides for thalli at different development stages can be obtained by using a continuous slide taking method according to growth periods of different thalli; and the accuracy for identification work of the filamentous fungus species is guaranteed.
Description
Technical field
The invention belongs to the mycology research field, be specifically related to a kind of flaking method that the filamentous fungus microscopic morphology is observed that is used for.
Background technology
For a long time, mainly be that morphological specificity with thalline is a foundation to the biological assay of filamentous fungus, therefore, obtaining preferably the thalline microscopic morphology, to observe slide just most important for accurate discriminating filamentous fungus kind.At present, the step of the traditional method of making filamentous fungus observation slide is following: at first with choosing the fungal cultures on the direct picking substratum of pin, place on the slide glass; Carefully the mycelium of picking is scatter covered with choosing pin then.But there is following significant disadvantages in this method: (1) is overlapping owing to thalline very easily appears in traditional flaking method; Be difficult to obtain observing effect slide preferably; Even skilled operator also need a large amount of film-makings, could obtain to meet the observation slide of observing requirement; (2) in choosing the sheet process; The nature of fungal spore on year spore body living status pole and is subject to artificial destruction; Therefore, tradition is chosen the observation slide that the sheet method obtains and often is difficult to reflect product spore mode and the type of fungi, and this characteristic the most key discriminating key element in the filamentous fungus classification just; (3) choose in the observation slide of sheet acquisition, thalline often is in development later stage, possibly obtain to grow early stage thalline hardly through the method for choosing sheet, and this is very unfavorable to research fungi different developmental phases morphological features.Therefore, just need set up a kind of new flaking method in the filamentous fungus field of identifying.
Summary of the invention
The purpose of this invention is to provide a kind of flaking method that the filamentous fungus microscopic morphology is observed that is used for; A kind of new flaking method that promptly is directed against the major defect of traditional flaking method and sets up; Be convenient to more comprehensively, accurately observe the microscopic morphology characteristic of filamentous fungus; Guaranteeing the accuracy of strain identification work, thereby remedy the deficiency of prior art.
Method of the present invention, its step is following: at first on water agar culture plate, take a groove, remove the substratum in the groove; The water agar piece of a size less than groove is placed in the groove, picking fungal cultures to be identified is inoculated in the water agar piece one side up that is positioned at groove, then the sterility cover slide is placed on the water agar piece again; Cover the petridish lid, cultivate under room temperature the sealing back, cultivates the end back and take out deckglass, is made into the observation slide of fungi.
The preparation method of above-mentioned water agar culture plate is following: after in 1000ml zero(ppm) water, adding 15 gram agar powders, 121 ℃, the 98.1kPa sterilization is poured in the sterile petri dish at last and is cooled off.
Above-mentioned groove is square, and its size is 1.6 * 1.6cm
2
The size of above-mentioned water agar piece is 0.4 * 0.4cm
2
Above-mentioned sterility cover slide size is 1.8 * 1.8cm
2
It is simple to operation that the thalline of method making filamentous fungus of the present invention is observed slide; Can be according to the growth cycle of different strain; Adopt the mode of getting sheet continuously, the thalline that obtains different development stage is observed slide, has guaranteed the accuracy of filamentous fungus species evaluation work.
Embodiment
Implementation step more specifically of the present invention:
(1) preparation water agar, autoclaving (121 ℃ are poured in the sterilized petridish (diameter 9cm) after 98.1kPa); The about 20ml of every ware; After treating its natural coagulation, on culture plate, take the square groove of 3-4 1.6 * 1.6cm, remove the substratum in the groove with scalper;
(2) remove before the interior substratum of groove, cut the water agar piece of 0.4 * 0.4cm with scalper, and be positioned over the groove central position;
(3) a little is inoculated in the water agar piece one side up of the 0.4 * 0.4cm that is positioned at the square groove center with choosing the pin picking will to have cultivated the fungal cultures to be identified of certain period;
(4) with sterilized size be 1.8 * 1.8cm deckglass lid on it, and make the culture block at groove center be positioned at deckglass central authorities, cover petridish lid and sealing then, under room temperature, cultivate;
(5) according to the growth characteristic of different sorts filamentous fungus, at the 3rd day that cultivates, the 6th day, the 10th day and the 15th day, take out 1-2 sheet deckglass respectively, select suitable floating supporting agent for use, make the observation slide, the morphological features that promptly can be used for observing different development stage.(1) of above-mentioned implementation step-(4) need be accomplished in super clean bench.
Through microscopic examination; Thalline on the deckglass of the filamentous fungus microscopic morphology of the inventive method preparation is in the self-sow state; The thalline overlapping phenomenon seldom appears; What is more important, because the self-sow state of thalline is not damaged, the observation slide of making is beneficial to the morphological specificity microscopic examination of (as producing spore mode and type).Mode through being employed in different incubation times taking-up deckglasses is made the observation slide; Realized continuous observation to the thalli growth development condition; Can obtain to reflect the observation slide of thalline different development stage morphological specificity comprehensively; Comprise that filamentous fungus produces the dynamic changing process of spore mode and sporulation, helps fungal species is accurately identified thereby grasp.
Claims (5)
1. one kind is used for the flaking method that the filamentous fungus microscopic morphology is observed, and it is characterized in that comprising the steps: at first on water agar culture plate, taking a groove, removes the substratum in the groove; The water agar piece of a size less than groove is placed in the groove, picking fungal cultures to be identified is inoculated in the water agar piece one side up that is positioned at groove, then the sterility cover slide is placed on the water agar piece again; Cover the petridish lid, cultivate under room temperature the sealing back, cultivates the end back and take out deckglass, is made into the observation slide of fungi.
2. flaking method as claimed in claim 1 is characterized in that the preparation method of described water agar culture plate is following: after in 1000ml zero(ppm) water, adding 15 gram agar powders, 121 ℃, the 98.1kPa sterilization is poured in the sterile petri dish at last and is cooled off.
3. flaking method as claimed in claim 1 is characterized in that described groove is square, and its size is 1.6 * 1.6cm
2
4. flaking method as claimed in claim 1, the size that it is characterized in that described water agar piece is 0.4 * 0.4cm
2
5. flaking method as claimed in claim 1 is characterized in that described sterility cover slide size is 1.8 * 1.8cm
2
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210001981.2A CN102533930B (en) | 2012-01-05 | 2012-01-05 | Manufacturing method of slide for observing filamentous fungus microscopic morphology |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210001981.2A CN102533930B (en) | 2012-01-05 | 2012-01-05 | Manufacturing method of slide for observing filamentous fungus microscopic morphology |
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| CN102533930A true CN102533930A (en) | 2012-07-04 |
| CN102533930B CN102533930B (en) | 2014-04-09 |
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| CN201210001981.2A Expired - Fee Related CN102533930B (en) | 2012-01-05 | 2012-01-05 | Manufacturing method of slide for observing filamentous fungus microscopic morphology |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103131748A (en) * | 2012-11-09 | 2013-06-05 | 黑龙江省农业科学院耕作栽培研究所 | Method for observing and identifying filamentous fungi |
| CN109628275A (en) * | 2019-01-11 | 2019-04-16 | 上海皓信生物科技有限公司 | The small culture apparatus of the improvement of filamentous fungi and its cultural method |
| CN109642859A (en) * | 2016-08-12 | 2019-04-16 | 樱花精机株式会社 | Coverslip sticker |
| CN109839380A (en) * | 2019-03-11 | 2019-06-04 | 江西农业大学 | A kind of pelletizer and method for filamentous fungi morphological feature microexamination |
| RU2719402C1 (en) * | 2019-12-30 | 2020-04-17 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Пермский государственный медицинский университет имени академика Е.А. Вагнера" Министерства здравоохранения Российской Федерации | Method of assessing the morphological structure of biofilms of microorganisms |
| CN111808911A (en) * | 2020-07-09 | 2020-10-23 | 江苏华绿生物科技股份有限公司 | Method for observing growth morphological structure of flammulina velutipes hyphae |
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| JPH03108499A (en) * | 1989-09-19 | 1991-05-08 | Shigeru Fujita | Culture medium for identifying fungus putting culture medium for many kind of fungi in interior of wall face of vessel having transparent and flat wall face |
| CN201501859U (en) * | 2009-09-15 | 2010-06-09 | 中国人民解放军第四军医大学 | Fungal microculture device capable of being directly observed under microscope |
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- 2012-01-05 CN CN201210001981.2A patent/CN102533930B/en not_active Expired - Fee Related
Patent Citations (2)
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| JPH03108499A (en) * | 1989-09-19 | 1991-05-08 | Shigeru Fujita | Culture medium for identifying fungus putting culture medium for many kind of fungi in interior of wall face of vessel having transparent and flat wall face |
| CN201501859U (en) * | 2009-09-15 | 2010-06-09 | 中国人民解放军第四军医大学 | Fungal microculture device capable of being directly observed under microscope |
Non-Patent Citations (4)
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| 《植物保护》 19900228 黄思良 "一种简便的真菌产孢状观察法" 20 1-5 , 第一期 * |
| 祖若夫: "《微生物学实验教程》", 30 April 1993, article ""实验6-1载片培养和真菌个体形态的观察"", pages: 429-430 * |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103131748A (en) * | 2012-11-09 | 2013-06-05 | 黑龙江省农业科学院耕作栽培研究所 | Method for observing and identifying filamentous fungi |
| CN109642859A (en) * | 2016-08-12 | 2019-04-16 | 樱花精机株式会社 | Coverslip sticker |
| CN109642859B (en) * | 2016-08-12 | 2021-05-18 | 樱花精机株式会社 | Cover glass pasting device |
| CN109628275A (en) * | 2019-01-11 | 2019-04-16 | 上海皓信生物科技有限公司 | The small culture apparatus of the improvement of filamentous fungi and its cultural method |
| CN109839380A (en) * | 2019-03-11 | 2019-06-04 | 江西农业大学 | A kind of pelletizer and method for filamentous fungi morphological feature microexamination |
| RU2719402C1 (en) * | 2019-12-30 | 2020-04-17 | Федеральное государственное бюджетное образовательное учреждение высшего образования "Пермский государственный медицинский университет имени академика Е.А. Вагнера" Министерства здравоохранения Российской Федерации | Method of assessing the morphological structure of biofilms of microorganisms |
| CN111808911A (en) * | 2020-07-09 | 2020-10-23 | 江苏华绿生物科技股份有限公司 | Method for observing growth morphological structure of flammulina velutipes hyphae |
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| CN102533930B (en) | 2014-04-09 |
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