UAS-Aβ42 flies were generated as previously described16 (link). Briefly, transgenic flies harboring UAS-Aβ42 were crossed with flies containing GMR-GAL4 to achieve eye-specific Aβ42 expression. Flies were bred on Jazz-Mix Drosophila fly food (Fisher Scientific, USA) containing 5 mM L-AIP or D-AIP, respectively. Five-day-old flies were collected and analyzed. Fly eyes were fixed in 2.5% glutaraldehyde, 2% formaldehyde in 100 mM PB buffer, pH 7.4, post-fixed in 1.5% osmium tetroxide. Samples were embedded in epon and polymerized at 60 °C for at least 48 hours. The sections were counterstained with aqueous uranyl acetate followed by lead citrate. Micrographs were taken using a Philips CM120 electron microscope at 80 kV and a 1 K CCD camera.
Cm120 electron microscope
Manufactured by Philips
Sourced in Netherlands, United States, France
The Philips CM120 is a transmission electron microscope designed for high-resolution imaging and analysis of samples. It features a 120 kV electron beam and provides advanced capabilities for visualizing and studying the microstructure of materials at the nanoscale level.
Automatically generated - may contain errors
Lab products found in correlation
Paraformaldehyde (pfa), by Electron Microscopy Sciences (8 mentions)
Beem capsules, by Electron Microscopy Sciences (4 mentions)
Megaview 3, by Philips (4 mentions)
Osmium tetroxide, by Merck Group (4 mentions)
Mega view 3 camera, by Olympus (4 mentions)
Item 5, by Olympus (4 mentions)
Spurr resin, by Electron Microscopy Sciences (3 mentions)
Digital micrograph, by Ametek (3 mentions)
High resolution ccd camera, by Ametek (2 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (2 mentions)
High resolution 2k x 2k ccd camera, by Ametek (2 mentions)
Propylene oxide, by Merck Group (2 mentions)
Sis morada digital camera, by Olympus (2 mentions)
Morada camera, by Olympus (2 mentions)
Embed 812, by Electron Microscopy Sciences (2 mentions)
Jazz mix drosophila fly food, by Thermo Fisher Scientific (1 mentions)
Orius 200 2kx2k camera, by Ametek (1 mentions)
Pinaacle 900t, by PerkinElmer (1 mentions)
Spectrum rx 1 ft ir spectrophotometer, by PerkinElmer (1 mentions)
Epon 812 epoxy resin, by Polysciences (1 mentions)
103 protocols using cm120 electron microscope
1
Drosophila Aβ42 Toxicity Analysis
2
Ultrastructural Analysis of Myelination
3
Immunolabeling Parasite Samples for TEM
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Preparations of parasites were fixed in 4% paraformaldehyde (Electron Microscopy Sciences, PA) in 0.25 M HEPES (pH 7.4) for 1 h at room temperature, then in 8% paraformaldehyde in the same buffer overnight at 4°C. They were infiltrated, frozen and sectioned as previously described [28 (link)]. The sections were immunolabeled with high affinity rat anti-HA monoclonal antibody 3F10 (1:50 in PBS/1% fish skin gelatin), then with anti-rat IgG antibodies, followed directly by 10 nm protein A-gold particles (Department of Cell Biology, Medical School, Utrecht University, the Netherlands) before examination with a Philips CM120 Electron Microscope (Eindhoven, the Netherlands) under 80 kV.
4
Ultrastructural Examination of Cells
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Cells were fixed with 2.5% glutaraldehyde in phosphate buffer and stored at 4°C until embedding. Cells were then post-fixed with 1% osmium tetroxide followed by increasing dehydration gradients of ethanol and acetone. Cells were then embedded in Araldite, and ultrathin sections were obtained (50–60 nm). Sections were collected onto uncoated copper grids, and stained with 3% lead citrate-uranyl acetate. Images were examined with a CM-120 electron microscope (Philips).
5
Zebrafish Tissue Fixation and Embedding
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Zebrafish embryos and larvae were fixed in 1.5% (v/v) glutaraldehyde and 1.5% (v/v) paraformaldehyde in 0.1 M cacodylate buffer (pH 7.4) several days at 4°C followed by washing in 0.1 M cacodylate buffer, 8% sucrose (10 minutes, 3 times). Then, samples were quickly washed in distilled water. After post-fixation in 1% osmium tetroxide in 0.1 M cacodylate buffer, 8% sucrose for 45 minutes at room temperature, samples were dehydrated in graded series of ethanol (30 to 100 %), 5 minutes each, and propylene oxide during 10 minutes. Finally, samples were embedded in epoxy resin. Ultrathin sections were stained with 7% uranyl acetate in methanol and lead citrate, and observed with a Philips CM120 electron microscope (Centre Technologique des Microstructures, Université Lyon 1, France) equipped with a Gatan Orius 200 2Kx2K camera.
For adult muscle tissues, 6-months trunk muscles were dissected in the fixative buffer under stereoscopic control and skin was delicately removed thanks to tweezers. Those thicker samples were treated as described above but with an extended sample incubation time (15 mins for each step). An additional 15 mins incubation in propylen oxide/ethanol 100% (v/v) followed by 2 X15 mins incubation in propylen oxide were performed. Times of incubation during the embedding steps were also prolonged.
For adult muscle tissues, 6-months trunk muscles were dissected in the fixative buffer under stereoscopic control and skin was delicately removed thanks to tweezers. Those thicker samples were treated as described above but with an extended sample incubation time (15 mins for each step). An additional 15 mins incubation in propylen oxide/ethanol 100% (v/v) followed by 2 X15 mins incubation in propylen oxide were performed. Times of incubation during the embedding steps were also prolonged.
6
Localization of T. gondii Rhoptry Protein
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Extracellular T. gondii endogenously expressing the C-terminal 3×HA tag (TgCA_RP-HA) was washed twice with PBS before fixation in 4% paraformaldehyde (Electron Microscopy Sciences, PA) in 0.25 M HEPES (pH 7.4) for 1 h at room temperature and then in 8% paraformaldehyde in the same buffer overnight at 4°C. Parasites were pelleted in 10% fish skin gelatin, and the gelatin-embedded pellets were infiltrated overnight with 2.3 M sucrose at 4°C and frozen in liquid nitrogen. Ultrathin cryosections were incubated in PBS and 1% fish skin gelatin containing mouse anti-HA antibody at a 1/5 dilution and then exposed to the secondary antibody that was revealed with 10-nm protein α-gold conjugates. Sections were observed and images were recorded with a Philips CM120 electron microscope (Eindhoven, the Netherlands) under 80 kV. For confirmation of rhoptry localization of TgCA_RP, ultrathin cryosections of T. gondii-infected cells were processed as described for extracellular parasites and immunolabeled with guinea pig anti-TgCA_RP and mouse anti-ROP7 antibodies at 1:25 and 1:750 dilutions, respectively, and detected with protein A-gold conjugates of 10 nm for TgCA_RP and 5 nm for ROP7.
7
TEM Imaging of Diluted Dispersions
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Transmission electron microscopy (TEM) images were obtained with a Phillips CM120 electron microscope (Eindhoven, Netherlands). One drop of highly diluted dispersion was placed onto a copper grid (mesh 200 and covered with formvar-carbon) and dried at room temperature before TEM analysis. The images were registered at 100 kV.
8
Autolysosome Formation after Steroid Starvation
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To assess autolysosome formation, after steroid starvation for 48 h, the cells were treated with dihydrotestosterone (DHT) for 3 days, and then washed twice with serum-free media. The cells were gently scraped, centrifuged and then fixed for 1 h at room temperature with 4% glutaraldehyde in cacodylate buffer (pH 7.0). Pellets were then embedded and sectioned for TEM analysis at 200 kV. Ultrathin sections were examined on a CM-120 electron microscope (Philips, Eindhoven, Netherlands).
9
Immunogold Labeling of Intracellular T. gondii
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Extracellular T. gondii parasites endogenously expressing the C-terminal 3×HA tag (TgZnT-HA) were washed twice with phosphate-buffered saline (PBS) before fixation in 4% paraformaldehyde (Electron Microscopy Sciences, PA) in 0.25 M HEPES (pH 7.4) for 1 h at room temperature and then in 8% paraformaldehyde in the same buffer overnight at 4°C. Parasites were pelleted in 10% fish skin gelatin, and the gelatin-embedded pellets were infiltrated overnight with 2.3 M sucrose at 4°C and frozen in liquid nitrogen. Ultrathin cryosections were incubated in PBS and 1% fish skin gelatin containing mouse anti-HA antibody at a 1/5 dilution and then exposed to the secondary antibody, which was revealed with 10-nm protein–anti-gold conjugates. Sections were observed, and images were recorded with a Philips CM120 electron microscope (Eindhoven, the Netherlands) under 80 kV.
10
Ultrastructural Analysis of Fungal Cell Walls
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Strains were grown statically from 1 × 107 conidia at 37°C in MM for 24 h. Mycelia were harvested and immediately fixed in 0.1 M sodium phosphate buffer (pH 7.4) containing 2.5% (vol/vol) glutaraldehyde and 2% (wt/vol) paraformaldehyde for 24 h at 4°C. Samples were encapsulated in agar (2%, wt/vol) and subjected to fixation (1% OsO4), contrasting (1% uranyl acetate), ethanol dehydration, and a two-step infiltration process with Spurr resin (Electron Microscopy Sciences) of 16 h and 3 h at RT. Additional infiltration was provided under vacuum at RT before embedment in BEEM capsules (Electron Microscopy Sciences) and polymerization at 60°C for 72 h. Semithin (0.5-μm) survey sections were stained with toluidine blue to identify the areas of best cell density. Ultrathin sections (60 nm) were prepared and stained again with uranyl acetate (1%) and lead citrate (2%). Transmission electron microscopy (TEM) images were obtained using a Philips CM-120 electron microscope at an acceleration voltage of 120 kV using a MegaView3 camera and iTEM 5.0 software (Olympus Soft Imaging Solutions GmbH). Cell wall thicknesses of 100 sections of different germlings were measured at ×23,500 magnification, and images were analyzed with the ImageJ software (85 (link)). Statistical differences were evaluated by using one-way analysis of variance (ANOVA) and Tukey’s post hoc test.
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