Mature ascomata were harvested from 30–50 day old cultures on OA. Ascomata were crushed and ascospores were picked using a dissecting needle and carefully transferred into distilled deionized water. A drop (5 μl) of this suspension was transferred to a polycarbonate membrane (1.0 Micron, 47 mm, GE Water and Process Technologies, Trevose, PA, USA). Polycarbonate membranes were placed on filter paper circles (0.7 mm, Schleicher & Schuell) to ensure that fluid was quickly absorbed through the pores of the membranes. The quick removal of fluid resulted in an equal distribution of ascospores and also more ascospores that could be viewed from the equatorial side as compared with passive evaporation of a droplet. The polycarbonate membranes with ascospore depositions were carefully cut out with a surgical knife and transferred to an aluminium stub. After drying at room temperature for one wk, the stubs were sputter-coated with gold three times for 30 s in a JEOL JFC-1300 Auto-fine coater and then viewed using a JEOL 5600LV scanning electron microscope (JEOL, Tokyo, Japan). Electron micrographs were acquired with the F4 scan at an acceleration voltage of 10 kV.
5600lv scanning electron microscope
Manufactured by JEOL
Sourced in Japan
The JEOL 5600LV is a scanning electron microscope (SEM) designed for high-resolution imaging and analysis of a wide range of samples. It features a low-vacuum mode that allows for the examination of non-conductive specimens without the need for extensive sample preparation. The 5600LV provides a magnification range of up to 300,000x and a spatial resolution of up to 3.5 nm, making it a versatile tool for various applications in material science, nanotechnology, and other fields.
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Lab products found in correlation
Oxford ct1500 cryostation, by JEOL (2 mentions)
Jfc 1300 auto fine coater, by JEOL (1 mentions)
Hummer 6.2 sputter coater, by Anatech (1 mentions)
Colloidal graphite, by Agar Scientific (1 mentions)
Smz25, by Nikon (1 mentions)
Ds ri2 camera, by Nikon (1 mentions)
Ax10 microscope, by Zeiss (1 mentions)
Nuclepore, by Cytiva (1 mentions)
16 protocols using 5600lv scanning electron microscope
1
SEM Imaging of Ascomycete Ascospores
2
cryo-SEM Analysis of Rhizopus-Burkholderia Interaction
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Low-temperature scanning electron microscopy of uncoated samples was performed as described previously (Schubert et al., 2007 (link); Jennessen et al., 2008 (link)). Fungal mycelium from the cured R. microsporus ATCC62417/S was inoculated on NA agar plate and incubated overnight to start hyphal growth. On the edges of hyphal spreading 2 µl of B. rhizoxinica culture was spotted and the plate was again incubated for approximately 2 hr. From the interaction zone several parts were selected and excised with a surgical blade as small agar blocks, and transferred to a copper cup for snap-freezing in nitrogen slush. Agar blocks were glued to the copper (KP-Cryoblock, Klinipath, Duiven, Netherlands). Samples were examined in a JEOL 5600LV scanning electron microscope (JEOL, Tokyo, Japan) equipped with an Oxford CT1500 Cryostation for cryo-electron microscopy (cryoSEM). Electron micrographs were acquired from uncoated frozen samples, or after sputter-coating by means of a gold/palladium target for three times during 30 s. Micrographs of uncoated samples were taken at an acceleration voltage of 3 kV, and consisted out of 30 averaged fast scans, and at 5 kV in case of the coated sample.
3
Scanning Electron Microscopy of Endothelial Cells
4
Cryogenic SEM of Fungal Colonies
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Wild-type, N402, and ΔtppA were grown for 1 week on AMM. Margins of colonies containing conidiophores were excised with a surgical blade and carefully transferred into a copper cup (diameter 10 mm, height 8 mm). Dislodging during snap freezing was prevented by gluing agar blocks in the copper cup with frozen tissue medium (KP-Cryoblock, Klinipath, Duiven, the Netherlands). The sample was snap-frozen in nitrogen slurry and immediately transferred in a vacuum transfer device to an Oxford CT1500 Cryostation attached to a JEOL 5600LV scanning electron microscope (JEOL, Tokyo, Japan). Electron micrographs were acquired from uncoated frozen samples, or after sputter-coating with gold three times during 30 s. Micrographs of uncoated samples were taken at an acceleration voltage of 2.5 kV, and consisted of 30 averaged fast scans (SCAN 2 mode). Coated samples were observed at 5 kV using F4 scans.
5
Cryogenic SEM Imaging of Acremonium
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The Acremonium isolate was cultured in microwells of MCC as described above. Different stages of growth and superstructure development were selected using a stereomicroscope and gently broken with a surgical blade. Microwells were transferred together with a piece of agar to a copper cup for snap-freezing in nitrogen slush. Agar blocks were glued to the copper surface with frozen tissue medium (KP-Cryoblock, Klinipath, Duiven, Netherlands). Samples were examined in a JEOL 5600LV scanning electron microscope (JEOL, Tokyo, Japan) equipped with an Oxford CT1500 Cryostation for cryo-SEM. Electron micrographs were acquired after sputter-coating using a gold target for three times during 30 s. Micrographs were taken at 5 kV and processed with Adobe Photoshop CS5.
6
SEM Microparticle Imaging Analysis
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SEM microparticle analyses were performed on a JEOL-5600 LV Scanning Electron Microscope (Tokyo, Japan) from JEOL, Japan, equipped with a SPRITE HR Four Axis Stage controller (Deben Research, Suffolk, UK). Samples were placed on metallic stubs covered with adhesive carbon tape (NEM tape, from Nisshin, Japan) and coated with gold/palladium using a Sputter Coater (Polaron, from Bad Schwalbach, Germany). All observations were performed under high vacuum with an acceleration voltage of 30 kV at a working distance of 12–13 mm and a spot size of 20.
7
Cryo-SEM Visualization of Cladosporium
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Isolates of Cladosporium spp. were grown on SNA with 30 g agar/L for 3–4 d at room temperature under black light. Relevant parts of the small colonies with conidiophores and conidia were selected under a binocular, excised with a surgical blade as small agar (3 × 3 mm) blocks, and transferred to a copper cup for snap-freezing in nitrogen slush. To prevent disruption of the intricate structure of the conidiophores by liquid nitrogen, a piece of Scotch tape was placed lightly over the opening of the copper cup. Agar blocks were glued to the copper surface with frozen tissue medium (KP-Cryoblock, Klinipath, Duiven, Netherlands) mixed with 1 part colloidal graphite (Agar Scientific, Stansted, UK). Samples were examined in a JEOL 5600LV scanning electron microscope (JEOL, Tokyo, Japan) equipped with an Oxford CT1500 Cryostation for cryo-electron microscopy (cryoSEM). Electron micrographs were acquired from uncoated frozen samples, or after sputter-coating by means of a gold target for three times during 30 s. Micrographs of uncoated samples were taken at an acceleration voltage of 3 kV, and consisted out of 30 averaged fast scans (SCAN 2 mode), and at 5 kV in case of the coated samples (PHOTO mode).
8
Scanning Electron Microscopy of Dried Mycelium
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Mycelium of two liquid static cultures was dried on top of each other at room temperature. Small rectangles (3 × 3 mm) were cut with a scalpel and attached with a 2 mm piece of Scotch tape in a 1 cm ø copper cup. After snap-freezing with liquid nitrogen, samples were transferred to a JEOL 5600 LV scanning electron microscope (JEOL, Tokyo, Japan) by the use of an Oxford CT1500 Cryostation. Ice was removed from the sample by sublimation at −85 °C. Gold was sputter coated for 2 min, after which micrographs were acquired at an acceleration voltage of 5 kV.
9
Preparation of Microbial Samples for SEM Analysis
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Hybrid and parent strains were cultivated on modified Dixon (mDixon) medium for 72 h, and a loop of cells was suspended in water. The cells were briefly vortexed to dislodge from each other. Droplets of 1, 2, and 3 μL were gently placed on mDixon agar and dried for 1 h in a laminar flow cabinet to fix the cells onto the agar. After preexamination under a stereomicroscope, small 4 × 4 mm selections with both individual cells and cells grouped together were cut out using a surgical blade (no. 11; Swann-Morton, Sheffield, UK) and glued on a copper sample cup with a small droplet of frozen-tissue medium (KP-Cryoblock; Klinipath, Duiven, The Netherlands) and subsequently snap-frozen in nitrogen slush, and transferred into an Oxford CT1500 Cryostation connected to a JEOL 5600LV scanning electron microscope (JEOL, Tokyo, Japan). Samples were sputter coated (3 × 1 min) using a gold target in the cryostation. Electron micrographs were taken at an acceleration voltage of 5 kV.
10
Cryo-SEM Imaging of Agar Samples
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Small rectangular 5- by 5-mm blocks of agar medium topped with a filter were transferred to copper cups for snap-freezing in nitrogen slush. They were glued to the copper surface with frozen tissue medium (KP-Cryoblock; Klinipath, Duiven, The Netherlands) and sputter coated 3 times for 1 min each by using a gold target. Cryo-scanning electron microscopy (cryo-SEM) was done with a JEOL 5600LV scanning electron microscope (JEOL, Tokyo, Japan) equipped with an Oxford CT1500 Cryostation. Electron micrographs were taken at an acceleration voltage of 5 kV.
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