CN112415033A - Application of double-conductive aluminum foil adhesive tape - Google Patents
Application of double-conductive aluminum foil adhesive tape Download PDFInfo
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- CN112415033A CN112415033A CN202011269614.1A CN202011269614A CN112415033A CN 112415033 A CN112415033 A CN 112415033A CN 202011269614 A CN202011269614 A CN 202011269614A CN 112415033 A CN112415033 A CN 112415033A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
- G01N23/2202—Preparing specimens therefor
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
- G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
- G01N23/225—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material using electron or ion
- G01N23/2251—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material using electron or ion using incident electron beams, e.g. scanning electron microscopy [SEM]
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2223/00—Investigating materials by wave or particle radiation
- G01N2223/07—Investigating materials by wave or particle radiation secondary emission
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2223/00—Investigating materials by wave or particle radiation
- G01N2223/10—Different kinds of radiation or particles
- G01N2223/102—Different kinds of radiation or particles beta or electrons
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2223/00—Investigating materials by wave or particle radiation
- G01N2223/60—Specific applications or type of materials
- G01N2223/612—Specific applications or type of materials biological material
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E60/00—Enabling technologies; Technologies with a potential or indirect contribution to GHG emissions mitigation
- Y02E60/10—Energy storage using batteries
Abstract
The invention provides an application of a double-conductive aluminum foil adhesive tape, and particularly relates to an application of a double-conductive aluminum foil adhesive tape in preparation of a filamentous fungus scanning electron microscope sample. The preparation method of the filamentous fungus scanning electron microscope sample comprises the following steps: 1. preparing a solid culture medium; 2. preparing a double-conductive aluminum foil adhesive tape; 3. inoculation and plug-in sheet; 4. climbing a double-conductive aluminum foil adhesive tape on a sample platform; 5. and (5) observing by a scanning electron microscope. The double-conductive aluminum foil adhesive tape has excellent conductivity, one side of the double-conductive aluminum foil adhesive tape is provided with the adhesive substance, aspergillus oryzae hyphae growing on the adhesive surface can be firmly attached to the adhesive substance, the hyphae is tightly attached to the double-conductive aluminum foil adhesive tape, the positions and the shapes of the hyphae, the sporangium and the spores on the carrier sheet cannot be changed, and the double-conductive aluminum foil adhesive tape has a better in-situ observation effect of electron microscope scanning.
Description
Technical Field
The invention relates to the technical field of scanning electron microscope detection of filamentous fungi, in particular to application of a double-conductive aluminum foil adhesive tape, and especially relates to application of the double-conductive aluminum foil adhesive tape in preparation of a scanning electron microscope sample.
Background
In situ observation of hyphae, sporangia and spores is an important means for performing hyphal morphology, sporangia morphological development and spore morphology of filamentous fungi. Although the traditional glass cover glass climbing method can observe the shapes of spores and hyphae under a light mirror, the glass sheet is not conductive when being observed by a scanning electron microscope, and the area of the glass sheet is large, so that the discharge is often caused to influence the quality of an observation result of the electron microscope, and therefore the glass cover glass on which the hyphae of the fungi climb is generally broken, the discharge phenomenon of the material is weakened, and the conductivity of the material is increased. However, this operation tends to destroy the original morphology of the mycelium and also to cause rupture of the mycelium and rupture of the sporangia, resulting in poor observation quality. Patent "CN 104928343 a" discloses a method for sample preparation using aluminum foil as a carrier, which has problems mainly: the surface of a common metal foil with conductivity is smooth, so that fungi hyphae are too short to climb onto an aluminum foil in a hypha growth stage, so that the quantity of the hyphae on the aluminum foil is small, and observation is not facilitated; in the sampling process, the aluminum foil needs to be moved, and mycelium is easy to separate from or slide off the smooth foil, so that an obvious trace exists during scanning of an electron microscope, and the observation result is influenced. Therefore, the above problems can be effectively solved by using a carrier which does not require separation to destroy mycelia and is advantageous for mycelia reprinting.
Disclosure of Invention
In view of the above technical problems, an object of the present invention is to provide an application of a double conductive aluminum foil tape, and in particular, an application of a double conductive aluminum foil tape in preparation of a scanning electron microscope sample. The double-conductive aluminum foil adhesive tape is used as a hypha climbing sheet carrier for the first time, the carrier sheet has excellent conductivity, one surface of the double-conductive aluminum foil adhesive tape is provided with a colloidal substance, aspergillus oryzae hyphae growing on the colloidal surface can be firmly attached to the surface, the hypha is tightly attached to the double-conductive aluminum foil adhesive tape, the positions and the shapes of the hypha, the sporangia and the spores on the carrier sheet cannot be changed, and the carrier sheet has a better in-situ observation effect of electron microscope scanning.
In order to achieve the purpose, the invention is realized by the following technical scheme:
an application of a double-conductive aluminum foil adhesive tape in preparation of a scanning electron microscope sample.
Preferably, the double-conductive aluminum foil adhesive tape is applied to preparation of filamentous fungi scanning electron microscope samples.
Preferably, the double-conductive aluminum foil adhesive tape is applied to the preparation of Aspergillus oryzae scanning electron microscope samples.
The preparation method of the filamentous fungus scanning electron microscope sample comprises the following steps:
(1) preparation of solid medium:
100mL DPY medium: 2g glucose, 1g peptone, 0.5g yeast extract, 0.5g KH2PO4,0.05g MgSO4Adjusting pH to 6.5-7.0, and adding 1.6g agar powder; sterilizing at 121 deg.C for 15min, and pouring into a glass culture dish with diameter of 9cm, wherein the thickness of the culture medium is 1-2 mm;
(2) preparing the double-conductive aluminum foil adhesive tape:
taking a double-conductive aluminum foil adhesive tape with the width of 15mm, cutting the double-conductive aluminum foil adhesive tape with the length of 15mm, and the area of 15mm multiplied by 15 mm; sterilizing at 121 deg.C for 20 min; after sterilization, carefully separating the double-conductive aluminum foil adhesive tape and the release paper from one corner of the double-conductive aluminum foil adhesive tape by using sterilized tweezers in an ultraclean sterile workbench for later use;
(3) inoculation and insertion:
taking 5 mu L of bacterium liquid spot to the center of a DPY culture medium by using a pipette in an ultra-clean sterile workbench; then, the double-conductive aluminum foil adhesive tape with the release paper removed is gently and obliquely inserted into a culture medium at a position 1cm away from the bacterial liquid point by using sterilized tweezers, and the adhesive surface of the double-conductive aluminum foil adhesive tape is close to the growing aspergillus oryzae bacterial colony to be used as a mycelium climbing surface; culturing in an incubator at 30 ℃ in the dark for 72-96 h, and stopping culturing when 2/3 of the double-conductive aluminum foil adhesive tape on the culture medium is covered by aspergillus oryzae hyphae;
(4) two electrically conductive aluminium foil adhesive tape climbing piece sample platform:
gently taking out the double-conductive aluminum foil adhesive tape, and adhering the surface without aspergillus oryzae hyphae growth to a sample table adhered with a conductive double-sided adhesive tape;
(5) and (3) observing by a scanning electron microscope:
and placing the sample stage adhered with the double-conductive aluminum foil adhesive tape slide in a scanning electron microscope for observation, and observing and photographing by using the scanning electron microscope under a vacuum mode and using a voltage of 30 kV.
The invention has the beneficial effects that:
(1) the double-conductive aluminum foil adhesive tape is used for replacing a glass cover glass, a sample can be entirely placed on a sample table without separating mycelium from a carrier, and the positions and the shapes of the mycelium, the sporangium and the spores which climb on the carrier sheet cannot be changed; (2) the double-conductive aluminum foil adhesive tape is adopted, so that the conductivity between an observation sample and a sample table is increased, the discharge phenomenon of the sample is reduced, and the sample is clearer under the same magnification; (3) one side of the double-conductive aluminum foil adhesive tape is provided with an adhesive substance, hyphae growing on the adhesive surface can be firmly attached to the adhesive substance, and the hyphae are tightly attached to the double-conductive aluminum foil adhesive tape, so that the double-conductive aluminum foil adhesive tape is particularly suitable for fungi with short hyphae, and a sample can obtain a good observation effect; (4) if the filamentous fungus sample is to be fixed, dehydrated and dried, and the mycelium and the carrier are not to be separated, the double conductive aluminum foil tape climber can be directly subjected to the relevant operation. Therefore, the double-conductive aluminum foil adhesive tape is adopted, and the development of hypha and sporangium and the growth state of spores can be better observed in situ.
Drawings
FIG. 1 shows the mycelium and sporangium of Control Aspergillus oryzae magnified 200X under a scanning electron microscope using a double conductive aluminum foil tape slide according to the present invention.
FIG. 2 shows the 200 Xmagnification of RNAi-IPC Aspergillus oryzae mycelia and sporangia under a scanning electron microscope using a double conductive aluminum foil tape slide according to the present invention.
FIG. 3A 500 Xmagnification of RNAi-IPC Aspergillus oryzae mycelia and sporangia under a scanning electron microscope using a double conductive aluminum foil tape slide according to the present invention.
FIG. 4 shows the mycelium and sporangium of Control Aspergillus oryzae in a scanning electron microscope magnified by 200X for a conventional aluminum foil slide.
FIG. 5 shows that 500 Xmagnification of the mycelium and the sporangium of RNAi-IPC Aspergillus oryzae was performed under a scanning electron microscope when the surface roughness of the aluminum foil was increased by 1500-mesh sandpaper and the aluminum foil was used as a carrier slide.
The invention is further explained with reference to the drawings and the embodiments.
Detailed Description
Example 1
An application of a double-conductive aluminum foil adhesive tape in preparation of filamentous fungi scanning electron microscope samples.
Wherein the filamentous fungus is Control Aspergillus oryzae.
The preparation method of the Aspergillus oryzae scanning electron microscope sample comprises the following steps:
(1) preparation of solid medium:
100mL DPY medium: 2g glucose, 1g peptone, 0.5g yeast extract, 0.5g KH2PO4,0.05g MgSO4Adjusting pH to 6.5-7.0, and adding 1.6g agar powder; sterilizing at 121 deg.C for 15min, and pouring into a glass culture dish with diameter of 9cm, wherein the thickness of the culture medium is 1-2 mm;
(2) preparing the double-conductive aluminum foil adhesive tape:
taking a double-conductive aluminum foil adhesive tape with the width of 15mm, cutting the double-conductive aluminum foil adhesive tape with the length of 15mm, and the area of 15mm multiplied by 15 mm; sterilizing at 121 deg.C for 20 min; after sterilization, carefully separating the double-conductive aluminum foil adhesive tape and the release paper from one corner of the double-conductive aluminum foil adhesive tape by using sterilized tweezers in an ultraclean sterile workbench for later use;
(3) inoculation and insertion:
taking 5 mu L of bacterium liquid spot to the center of a DPY culture medium by using a pipette in an ultra-clean sterile workbench; then, the double-conductive aluminum foil adhesive tape with the release paper removed is gently and obliquely inserted into a culture medium at a position 1cm away from the bacterial liquid point by using sterilized tweezers, and the adhesive surface of the double-conductive aluminum foil adhesive tape is close to the growing aspergillus oryzae bacterial colony to be used as a mycelium climbing surface; culturing in an incubator at 30 ℃ in the dark for 72-96 h, and stopping culturing when 2/3 of the double-conductive aluminum foil adhesive tape on the culture medium is covered by aspergillus oryzae hyphae;
(4) two electrically conductive aluminium foil adhesive tape climbing piece sample platform:
gently taking out the double-conductive aluminum foil adhesive tape, and adhering the surface without aspergillus oryzae hyphae growth to a sample table adhered with a conductive double-sided adhesive tape;
(5) and (3) observing by a scanning electron microscope:
and placing the sample stage adhered with the double-conductive aluminum foil adhesive tape slide in a scanning electron microscope for observation, and observing and photographing by using the scanning electron microscope under a vacuum mode and using a voltage of 30 kV. The observation results are shown in FIG. 1.
Example 2
Wherein the filamentous fungus is RNAi-IPC Aspergillus oryzae. The rest is the same as example 1. The observation results are shown in FIGS. 2 and 3.
Comparative example 1
Wherein, the ordinary aluminum foil is adopted to replace the double-conductive aluminum foil adhesive tape as the climbing sheet carrier, and the rest is the same as the embodiment 1. The observation results are shown in FIG. 4.
Comparative example 2
Wherein, the general aluminum foil treated by 1500-mesh sand paper is used as the carrier of the climbing sheet, and the rest is the same as the embodiment 1. The observation results are shown in FIG. 5.
FIG. 1 shows the mycelium and sporangium of Control Aspergillus oryzae magnified 200X under a scanning electron microscope using a double conductive aluminum foil tape slide according to the present invention. FIG. 2 shows the 200 Xmagnification of RNAi-IPC Aspergillus oryzae mycelia and sporangia under a scanning electron microscope using a double conductive aluminum foil tape slide according to the present invention. FIG. 3A 500 Xmagnification of RNAi-IPC Aspergillus oryzae mycelia and sporangia under a scanning electron microscope using a double conductive aluminum foil tape slide according to the present invention. As can be seen from FIGS. 1-3, the observation results of the filamentous fungus scanning electron microscope samples prepared in the examples are better, the mycelium and the sporangia can be clearly observed, and the mycelium and the sporangia are distributed on the slide-climbing carrier more, which is beneficial to observation.
FIG. 4 shows the mycelium and sporangium of Control Aspergillus oryzae in a scanning electron microscope magnified by 200X for a conventional aluminum foil slide. When the slide is taken down from the culture medium, the number of mycelia is too small during observation due to the fact that the surface of a common aluminum foil is smooth and the mycelia easily slide, and meanwhile, dark marks are left at the positions where the mycelia slide, so that the scanning result is extremely influenced. In particular, some Aspergillus oryzae after genetic engineering modification has extremely short hypha, and is easy to fall off from smooth aluminum foil when climbing sheets are taken.
As can be seen from FIG. 5, when a common aluminum foil is used as a carrier, 1500-mesh sand paper is used to increase the surface roughness of the aluminum foil and increase the adhesion of mycelia, but when the scanning magnification of an electron microscope is increased to 500X, scratches formed by the sand paper have very obvious scratches under the scanning electron microscope, and some scratches are similar to the shape of the mycelia, thus extremely interfering the observation of the mycelia.
Finally, it should be emphasized that the above-described preferred embodiments of the present invention are merely examples of implementations, rather than limitations, and that many variations and modifications of the invention are possible to those skilled in the art, without departing from the spirit and scope of the invention.
Claims (3)
1. An application of a double-conductive aluminum foil adhesive tape in preparation of a scanning electron microscope sample.
2. Use according to claim 1, wherein the scanning electron microscope sample comprises a filamentous fungus scanning electron microscope sample.
3. The use according to claim 2, wherein the preparation method of the filamentous fungus scanning electron microscope sample comprises the following steps:
(1) preparation of solid medium:
100mL DPY medium: 2g glucose, 1g peptone, 0.5g yeast extract, 0.5g KH2PO4,0.05g MgSO4Adjusting pH to 6.5-7.0, and adding 1.6g agar powder; sterilizing at 121 deg.C for 15min, and pouring into a glass culture dish with diameter of 9cm, wherein the thickness of the culture medium is 1-2 mm;
(2) preparing the double-conductive aluminum foil adhesive tape:
taking a double-conductive aluminum foil adhesive tape with the width of 15mm, cutting the double-conductive aluminum foil adhesive tape with the length of 15mm, and the area of 15mm multiplied by 15 mm; sterilizing at 121 deg.C for 20 min; after sterilization, carefully separating the double-conductive aluminum foil adhesive tape and the release paper from one corner of the double-conductive aluminum foil adhesive tape by using sterilized tweezers in an ultraclean sterile workbench for later use;
(3) inoculation and insertion:
taking 5 mu L of bacterium liquid spot to the center of a DPY culture medium by using a pipette in an ultra-clean sterile workbench; then, the double-conductive aluminum foil adhesive tape with the release paper removed is gently and obliquely inserted into a culture medium at a position 1cm away from the bacterial liquid point by using sterilized tweezers, and the adhesive surface of the double-conductive aluminum foil adhesive tape is close to the growing aspergillus oryzae bacterial colony to be used as a mycelium climbing surface; culturing in an incubator at 30 ℃ in the dark for 72-96 h, and stopping culturing when 2/3 of the double-conductive aluminum foil adhesive tape on the culture medium is covered by aspergillus oryzae hyphae;
(4) two electrically conductive aluminium foil adhesive tape climbing piece sample platform:
gently taking out the double-conductive aluminum foil adhesive tape, and adhering the surface without aspergillus oryzae hyphae growth to a sample table adhered with a conductive double-sided adhesive tape;
(5) and (3) observing by a scanning electron microscope:
and placing the sample stage adhered with the double-conductive aluminum foil adhesive tape slide in a scanning electron microscope for observation, and observing and photographing by using the scanning electron microscope under a vacuum mode and using a voltage of 30 kV.
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| CN202011269614.1A CN112415033B (en) | 2020-11-13 | 2020-11-13 | Application of double-conductive aluminum foil adhesive tape |
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