After culturing hCECs on culture inserts, cells were fixed with 2% glutaraldehyde/2% PFA in phosphate-buffered saline (PBS) for 2 h at 4 °C. After washing three times with PBS, it was dehydrated with an ethanol series (30%, 50%, 70%, 80%, 90%, 95%, and 99.5% ethanol) and further treated with t-butyl alcohol. Cells on culture inserts were dried in a freeze-drying device (JFD-320; JEOL Ltd., Tokyo, Japan). Then, the bottom side of the membrane of the culture insert was adhered to the sampling stage with carbon tape. The surrounding membrane was cut off using disposable biopsy punches resulting in removal of the surrounding plastic container of the insert. The specimen pasted onto the sampling stage was subjected to platinum coating using an auto fine coater (FCL8 1600; JEOL) for 30 s. Then, the sample was observed by SEM (JSM-6510LA; JEOL) at 20 kV.
Jfd 320
Manufactured by JEOL
Sourced in Japan
The JFD-320 is a field emission scanning electron microscope (FE-SEM) designed for high-resolution imaging of a wide range of materials. It features a high-brightness electron gun and advanced optics to deliver exceptional image quality and resolution. The JFD-320 is capable of operating at accelerating voltages between 0.5 and 30 kV, allowing for versatile imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Jsm 6010la, by JEOL (5 mentions)
Jfc 1600, by JEOL (5 mentions)
Jsm 6510, by JEOL (3 mentions)
Jee 4x, by JEOL (2 mentions)
Jsm 6510la, by JEOL (1 mentions)
Neoscope jcm 5000, by JEOL (1 mentions)
Jsm 6380lv, by JEOL (1 mentions)
Jsm 7900f, by JEOL (1 mentions)
Jcm 5000, by JEOL (1 mentions)
Jem 6490lv, by JEOL (1 mentions)
Jsm 6510lv scanning electron microscope, by JEOL (1 mentions)
Tert butanol, by Merck Group (1 mentions)
Jfc 1100, by JEOL (1 mentions)
Jsm 6510lv, by JEOL (1 mentions)
Sc7620 sputter coater, by Quorum Technologies (1 mentions)
Jsm 6390lv, by JEOL (1 mentions)
Jsm 5600lv, by JEOL (1 mentions)
Model h 7100, by Hitachi (1 mentions)
Ve 9800, by Keyence (1 mentions)
Jfcc 160 ion sputterer, by JEOL (1 mentions)
25 protocols using jfd 320
1
SEM Imaging of Cultured Human Corneal Endothelial Cells
2
Scanning Electron Microscopy of Testicular Cells
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Suspensions of testicular cells or mature sperm from the sperm duct were immobilized on a poly L-lysine-coated coverslip. Samples were fixed in 2.5% glutaraldehyde in 0.45 M sucrose and 0.1 M sodium cacodylate (pH 7.4) at 4°C for 1 h. Fixed samples were washed three times with 0.1 M sodium cacodylate (pH 7.4), dehydrated using a graded ethanol series, substituted with t-butyl alcohol and freeze-dried (JFD-320, JEOL, Tokyo, Japan), coated with Au using an ion sputter gun, and observed under a scanning electron microscope (NeoScope JCM-5000, JEOL).
3
Specimen Preparation and Morphological Analysis of Wasps
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Adults were killed in 99.5% (v/v) ethanol and dried following the t-butyl-alcohol freeze-drying method (originally developed by Inoué and Osatake 1988 (link)) using a freeze-drying device (JFD-320, JEOL, Tokyo, Japan) following the manufacturer’s protocol. The specimens examined were deposited in the NSMT.
The external structures of dry-mounted wasps were examined using binocular stereomicroscopes (S8APO and MZ APO, Leica Microsystems KK, Tokyo, Japan) fitted with digital single-lens reflex cameras (E-30, Olympus, Tokyo, Japan) and a scanning electron microscope (JSM-6380LV, JEOL, Tokyo, Japan) operating at 1.5 kV. The length of each body part was measured using an ocular micrometer.
The following morphological abbreviations were used: POL, postocellar line (the distance between the inner edges of the two lateral ocelli); OOL, ocular-ocellar line (the distance from the outer edge of a lateral ocellus to the compound eye); and LOL, lateral-ocellar line (the distance between the median and lateral ocelli). The morphological terminology follows Richards (1977) , Ronquist and Nordlander (1989) , and Liljeblad et al. (2008) ; the description of surface sculptures follows Harris (1979) .
The external structures of dry-mounted wasps were examined using binocular stereomicroscopes (S8APO and MZ APO, Leica Microsystems KK, Tokyo, Japan) fitted with digital single-lens reflex cameras (E-30, Olympus, Tokyo, Japan) and a scanning electron microscope (JSM-6380LV, JEOL, Tokyo, Japan) operating at 1.5 kV. The length of each body part was measured using an ocular micrometer.
The following morphological abbreviations were used: POL, postocellar line (the distance between the inner edges of the two lateral ocelli); OOL, ocular-ocellar line (the distance from the outer edge of a lateral ocellus to the compound eye); and LOL, lateral-ocellar line (the distance between the median and lateral ocelli). The morphological terminology follows Richards (1977) , Ronquist and Nordlander (1989) , and Liljeblad et al. (2008) ; the description of surface sculptures follows Harris (1979) .
4
Morphological Characterization of Fungal Cells
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Controls and baicalein-, wogonin-, or TPEN-treated T. rubrum, T. mentagrophytes, A. fumigatus, and C. albicans cells were grown on Sabouraud dextrose agar medium or in Sabouraud liquid medium for 24 h at 28 °C. The treated cells were collected in conical centrifuge tubes and fixed with 2.5% (v/v) glutaraldehyde in 0.1 M phosphate buffer (PB; pH 7.3) at 4 °C for 2 h. This was followed by five consecutive washes with 0.1 M PB, post-fixing with 1% (v/v) osmium tetroxide in 0.1 M PB at 4 °C for 2 h, and five consecutive washes with 0.1 M PB. The treated fungal cells were incubated with an aqueous solution of 1% (w/v) tannin (air-free) at 4 °C in the dark for 1 h and washed five times with 0.1 M PB. Samples were mixed with 1% (v/v) osmium tetroxide in 0.1 M PB x at 4 °C for 1 h and dehydrated in a graded EtOH series. The dehydrated samples were immersed five times in t-butyl alcohol and lyophilised by vacuum freeze-drying (JFD-320; JEOL, Tokyo, Japan). Samples were mounted using silver paste (Nisshin EM, Tokyo, Japan), sputter-coated with gold using an ion coater (VX-10A; Eiko, Tokyo, Japan), and monitored by SEM in the voltage range of 5–15 kV (JSM-6510; JEOL).
5
Root Tip Imaging via SEM
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Approximately 1 cm root tips were excised and used for scanning electron microscopy (SEM) analysis to image the surface of root tips as described by Ahmed et al. (2017) (link). Briefly, the root tips were fixed in 0.1 M sodium phosphate buffer (pH 7.2) with 2% paraformaldehyde and 2.5% glutaraldehyde for 2 d at 0–4 °C. The root tips were then dehydrated through a gradient series of ethanol (30%-50%-70%-80%-90%-95%, once for 15 min at each step). The root tips were then freeze-dried using a freeze-dryer (JEOL JFD-320, Tokyo, Japan). Dehydrated roots were then put on two-sided carbon tape fixed on an SEM stub and visualized under a JEM-6490 LV scanning electron microscope (JEOL, Tokyo, Japan). The experiment was repeated three times with 4 root tips in each replicate.
6
Analyzing Gardnerella Biofilm Formation
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SEM was used to observe the formation of Gardnerella species biofilms in a medium at different pH values. The ATCC14018 suspension was adjusted to a final concentration of 106 CFU/ml in sBHI at different pH values. The samples were cultured in 24-well plates with a 14 ×14 mm circular cover glass at the bottom of each well at 37°C under anaerobic conditions without agitation for 48 h. The sBHI in each well was removed lightly, and the samples in the well were then washed once with 300 μl 1 × PBS. The planktonic bacteria of each sample were removed and the adhered biofilm was left in the wells. The samples were fixed with 3% glutaraldehyde at 4°C overnight, dehydrated using an ethanol series, and substituted with t-butyl alcohol for freeze-drying (JFD-320, JEOL, Japan). The dried specimen was coated with platinum using an auto fine coater (JFC-3000EC, JEOL, Japan) and observed using SEM (JSM- 7900F, JEOL, Japan) at 3 kV (Thellin et al., 2016 (link); Jung et al., 2019 (link)).
7
Electron Microscopy Sample Preparation
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Samples were fixed and post-fixed by the same procedure used for transmission electron microscopy, dehydrated in a 30–100% EtOH series, substituted by t-butanol and frozen at −30 °C. Samples were then lyophilized in a freeze-drying device (JFD-320, JEOL), mounted on an aluminum block and sputter coated with Au-Pd using an Auto Fine Coater JEC-3000FC (JEOL). Samples were observed under a SEM (JCM5000, JEOL).
8
Morphological Analysis of Flag Leaf
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On the day of flowering, the main stems of five random plants with uniform growth were selected and marked. Ten days after flowering, the entire blade of flag leaves was cut and immediately placed in a pre-cooled 3% glutaraldehyde fixative solution (in 0.1 M phosphate-buffered saline [PBS], Ph = 7.0) and stored in a refrigerator at 4 °C. The fixed leaves were washed three times with 0.1 M PBS, and samples were cut from the leaf tip, leaf center, and leaf base. Then, the samples were dehydrated with a gradient ethanol series of 30%, 50%, 70%, 80%, 90%, and 100% (twice) for 15 min each, followed by 75% tert-butanol (in ethanol solution) and 100% tert-butanol (twice) for 20 min each. The dehydrated samples were freeze-dried (JFD-320; JEOL Ltd., Akishima, Tokyo, Japan), stuck on stubs, and coated with platinum by using an ion sputtering device (JEM-6490LV; JEOL). The prepared samples were observed under a scanning electron microscope (JEM-6490LV; JEOL).
9
Scanning Electron Microscopy of Dehydrated PMCs
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PMCs were dehydrated in a series of alcohols of increasing concentrations (from 50 up to 100%) for 20 min at each stage. Then they were suspended in tert-butanol (Sigma-Aldrich) two times for 20 min each at 26 °C. Further on, the PMCs were held in tert-butanol for 12 h at 4 °C. The specimens were then freeze-dried (JFD-320, JEOL, Japan). An adhesive tape was used for the preparation of open capsules.
The dried specimens of both types were fixed on aluminium disks by means of a current-conducting tape and coated with gold using a vacuum sputtering equipment JFC-1100 (JEOL, Japan) for better resolution. The specimens were examined in a JSM-6510LV scanning electron microscope (JEOL, Japan).
The dried specimens of both types were fixed on aluminium disks by means of a current-conducting tape and coated with gold using a vacuum sputtering equipment JFC-1100 (JEOL, Japan) for better resolution. The specimens were examined in a JSM-6510LV scanning electron microscope (JEOL, Japan).
10
Specimen Preparation for SEM Imaging
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The specimens were immersed in half-strength Karnovsky’s solution (2% paraformaldehyde, 2.5% glutaraldehyde, pH 7.4) for 30 min. The fixed specimens were dehydrated using 50, 70, 80, 90, 95 and 100% (v/v) graded ethanol, and transferred into t-butyl alcohol. The specimens were then freeze-dried (JFD-320; JEOL, Tokyo, Japan). The samples were coated with platinum by sputtering with a plasma multicoater (PMC-5000; Meiwafosis, Tokyo, Japan) (Asahi et al. 2012 (link)). Samples that were treated with the conventional method were used as controls.
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