Flies were prepared for transmission electron microscopy essentially as described in Longley and Ready (1995) (link) with modifications. Ultrathin sections were cut with a diamond knife using a Reichert-Jung Ultracut microtome. Sections were stained with Reynold's lead citrate and 2% aqueous uranyl acetate and were observed with a Philips CM-100 transmission electron microscope. For electron tomography, 420 nm thick sections were cut and a 200 kV Tecnai G2 20 electron microscope (FEI) equipped with a Gatan CCD camera was used to record serial tilts of ±60° in increments of 1°. Tomogram reconstruction was carried out using IMOD software (Kremer et al., 1996 (link)).
Tecnai g2 20 electron microscope
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Tecnai G2 20 electron microscope is a high-resolution transmission electron microscope (TEM) designed for advanced imaging and analysis of materials at the nanoscale. It features a 200 kV field-emission electron source and a range of advanced imaging and analytical capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Ultracut e ultramicrotome, by Reichert Technologies (5 mentions)
Us1000 2k ccd camera, by Thermo Fisher Scientific (2 mentions)
Em uc6 ultramicrotome, by Leica (2 mentions)
Escalab 250xi, by Thermo Fisher Scientific (2 mentions)
Ccd camera, by Ametek (1 mentions)
Ultracut microtome, by Reichert Technologies (1 mentions)
Cm100 transmission electron microscope, by Philips (1 mentions)
D8 advance x ray powder diffractometer, by Bruker (1 mentions)
Uv 2450, by Shimadzu (1 mentions)
Lead citrate, by Merck Group (1 mentions)
Ultracut s, by Leica (1 mentions)
Em pact2, by Leica (1 mentions)
Quetol 651 resin, by Nisshin EM (1 mentions)
Thiocarbohydrazide, by Thermo Fisher Scientific (1 mentions)
Ultracut s ultramicrotome, by Leica (1 mentions)
Quetol 651, by Nisshin EM (1 mentions)
Helios g4 cx, by Thermo Fisher Scientific (1 mentions)
Miniflex 600, by Rigaku (1 mentions)
Dfc320 camera, by Leica (1 mentions)
Jsm 6700f scanning electron microscope, by JEOL (1 mentions)
14 protocols using tecnai g2 20 electron microscope
1
Transmission Electron Microscopy of Flies
2
Comprehensive Characterization of Materials
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A Bruker D8-Advance powder X-ray diffractometer with Cu-Kα radiation (λ = 1.5418 Å) was used to conduct the Powder X-ray Diffraction (PXRD) investigations. A Nicolet Protege 460 Fourier transform infrared (FT-IR) spectrometer was used to conduct FT-IR spectroscopy measurements utilizing KBr as the standard of transmission within the 400–4000 cm−1 range. With the FEI Tecnai G2 20 electron microscope running at a 200 kV accelerating voltage, studies using transmission electron microscopy (TEM) as well as energy dispersive X-ray analysis (EDX) were conducted. Employing FEI Quanta 3D FEG/FESEM at an accelerating voltage of 20 kV, field emission scanning electron microscopy (FESEM) was conducted utilizing gold-coated discs. Using barium sulphate as a reference, the diffuse reflectance spectra of the samples were captured using the UV-Visible spectrophotometer Shimadzu UV-2450 in the wavelength range of 200–800 nm. Using a Nova 2000e Surface Area and Pore Size Analyzer (Quantachrome Instruments), nitrogen adsorption–desorption isotherms were recorded at liquid nitrogen temperature (77 K), and the specific area was calculated using the Brunauer–Emmett–Teller (BET) technique. Before the surface area measurements, the samples were degassed for 12 hours at 150 °C.
3
Ultrastructural Analysis of SINV-Infected Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BHK-15 cells infected with wild-type or mCherry-E2-tagged SINV at an MOI 5 were fixed at 6 or 12 h p.i. Cells were fixed for three days in 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer, embedded in 2% agarose, post-fixed for 90 min in buffered 1% osmium tetroxide containing 0.8% potassium ferricyanide, and stained for 45 min in 2% uranyl acetate. They were then dehydrated with a graded series of ethanol, transferred into propylene oxide and embedded in EMbed-812 resin. Thin sections were cut on a Reichert-Jung Ultracut E ultramicrotome and stained with 2% uranyl acetate and lead citrate [37 (link)]. Images were acquired in an FEI Tecnai G2 20 electron microscope equipped with a LaB6 source and operated at 100 keV (Life Science Microscopy Facility, Purdue University, West Lafayette, IN, USA).
4
Ultrastructural Analysis of SINV Infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BHK-15 and C6/36 cells infected with wild-type or mCherry-E2-tagged SINV at an MOI of 5 were fixed at 6, 12, or 24 h p.i. Persistently infected C6/36 cells were fixed at 48 h after seeding. Cells were fixed for 3 days in 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer, embedded in 2% agarose, postfixed for 90 min in buffered 1% osmium tetroxide containing 0.8% potassium ferricyanide, and stained for 45 min in 2% uranyl acetate. They were then dehydrated with a graded series of ethanol, transferred into propylene oxide, and embedded in EMbed-812 resin. Thin sections were cut on a Reichert-Jung Ultracut E ultramicrotome and stained with 2% uranyl acetate and lead citrate (30 (link)). Images were acquired in an FEI Tecnai G2 20 electron microscope equipped with a LaB6 source and operated at 100 keV (Life Science Microscopy Facility, Purdue University).
5
Nanowire-Cell Interaction Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Once the cells were incubated with the wires for a given amount of time, the nanowire solution was removed via pipette and rinsed with 1 mL of PBS. The PBS was replaced with 2 mL of 2.5% glutaraldehyde and 0.1 M sodium cacodylate buffer solution (fix). This solution was allowed to sit for several minutes before being poured off. Another 2 mL of the fix was added and the cells were scraped from the bottom of the dish and transferred to a small conical centrifuge tube. The cells were spun down, the old fix was removed and 1.5 mL of fresh fix was added.
The remainder of the processing was done at the Purdue Life Sciences Microscopy Facility. Cells were embedded in 2% agarose, and post-fixed in buffered 1% osmium tetroxide containing 0.8% potassium ferricyanide. Cells were then en bloc stained in 4% uranyl acetate, dehydrated with a graded series of ethanol, and embedded in LX-112 resin. Sections with a 90 nm thickness for the 37 °C and 180 nm for the 4 °C samples were cut using a Reichert-Jung Ultracut E ultramicrotome and stained with 2% uranyl acetate and lead citrate. Images were acquired on a FEI Tecnai G220 electron microscope equipped with a LaB6 source and operated at 100 kV.
The remainder of the processing was done at the Purdue Life Sciences Microscopy Facility. Cells were embedded in 2% agarose, and post-fixed in buffered 1% osmium tetroxide containing 0.8% potassium ferricyanide. Cells were then en bloc stained in 4% uranyl acetate, dehydrated with a graded series of ethanol, and embedded in LX-112 resin. Sections with a 90 nm thickness for the 37 °C and 180 nm for the 4 °C samples were cut using a Reichert-Jung Ultracut E ultramicrotome and stained with 2% uranyl acetate and lead citrate. Images were acquired on a FEI Tecnai G220 electron microscope equipped with a LaB6 source and operated at 100 kV.
6
High-Pressure Freezing and Electron Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A cell pellet was harvested from approx. 300 ml of two-week-old culture by centrifuging at 1,200 x g for 4 min, and then was high-pressure frozen (at -196 °C under 2,181 bar) using a Leica EM PACT2 (Leica). The frozen pellet was transferred to 1% osmium tetroxide in HPLC grade acetone and substituted for 118 hours at -85 °C, then warmed to -20 °C over five hours. After this, the specimen was warmed to 4 °C over 48 hours and then washed in acetone. Subsequently, the specimen was transferred through increasing concentrations of Quetol 651 resin (Nisshin EM Co., Ltd.) in acetone: 1:2 resin/acetone for 1 hr, 1:1 for 2 hr, and 2:1 for 3 hr. The specimen was then embedded in pure Quetol 651 resin overnight at room temperature, and this step repeated again with fresh resin. The embedded specimen was polymerized overnight in fresh resin at 65 °C.
Ultrathin sections (ca. 70 nm) of both EM blocks were prepared using an ultramicrotome (Ultracut S; Leica), double stained with 2.0% uranyl acetate and 2.0% lead citrate (Sigma-Aldrich Co.), and observed in a Tecnai G2 20 electron microscope (FEI).
Ultrathin sections (ca. 70 nm) of both EM blocks were prepared using an ultramicrotome (Ultracut S; Leica), double stained with 2.0% uranyl acetate and 2.0% lead citrate (Sigma-Aldrich Co.), and observed in a Tecnai G2 20 electron microscope (FEI).
7
Ultrastructural Analysis of Marine Organism Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For egg sections, fixed eggs were washed with filtered (pore size = 0.2 µm) artificial seawater (Rohto Marine, Iwaki, Tokyo, Japan) (FASW) and post-fixed with 2.0% osmium tetroxide dissolved in FASW for 2 h at 4°C. After washing with 8.0% sucrose aqueous solution, the conductive staining was performed by incubating 0.5% thiocarbohydrazide (Thermo Fisher Scientific, Waltham, MA, USA) aqueous solution for 30 min and 1.0% osmium tetraoxide aqueous solution for 1 h at 4°C. The samples were washed, dehydrated in a graded series (30, 50, 70, 80, 90 and 100%) of N,N-dimethylformamide, cleared in n-butyl-glycidyl-ether (Nisshin EM, Tokyo, Japan) and embedded in Quetol 651 (Nisshin EM). Ultra-thin sections (60 nm thickness) were cut with a diamond knife on an Ultracut S ultra-microtome (Leica Microsystems, Wetzlar, Germany), stained with 2.0% uranyl acetate solution and 2.0% lead citrate solution, and observed using a Tecnai G2 20 electron microscope (FEI, Hillsboro, OR, USA) operated at 120 kV. Thin sections of the ovary and gill were prepared as described above with modification in the dehydration process by using a graded series (30, 50, 70, 80, 90 and 100%) of ethanol.
8
Characterization of TiO2-CuOx Thin Films
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Surface morphology and composition of received TiO2–CuOx layers were investigated by scanning electron microscopy Helios G4 CX (FEI, Hillsboro, Oregon, USA) and energy-dispersive X-ray (EDX) analysis. The study of suspension particle aggregates was carried out using Tecnai G2 20 electron microscope (FEI, Hillsboro, Oregon, USA), at an accelerating voltage of 200 kV. The microscope is equipped with a system for energy-dispersive X-ray analysis.
The phase composition of the obtained TiO2–CuOx layers was studied using X-ray diffraction. Diffraction wide-angle spectra were recorded by Miniflex 600 (Rigaku, Tokyo, Japan) under the following conditions:
The phase composition of the obtained TiO2–CuOx layers was studied using X-ray diffraction. Diffraction wide-angle spectra were recorded by Miniflex 600 (Rigaku, Tokyo, Japan) under the following conditions:
Scanning speed 10 °C·min−1;
Scan range from 3° to 90°;
Radiation Cu Kα;
The voltage on the tube 20 Kv;
The anode current 7 mA.
9
Ultrastructural Analysis of Sol Muscle
10
Ultrastructural Analysis of Vocal Fold Epithelium
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!