Ten microliters of the CsCl-purified phage suspension (109 PFU/mL) were applied to 400-mesh carbon-coated copper grids, negatively stained with 1% uranyl acetate and analyzed using a JEM-100C (JEOL, Akishima, Japan) transmission electron microscope at 80 kV accelerating voltage. Images were taken on Kodak film SO-163 (Kodak, Cat. \# 74144, Hatfield, PA, USA). Phage particle dimensions were measured using ImageJ version 1.53e [https://imagej.nih.gov/ij/index.html , accessed on 1 June 2021] in relation to the scale bar generated from the microscope [14 ].
So 163 film
Manufactured by Kodak
Sourced in United States, Japan
The SO-163 film is a black-and-white film product manufactured by Kodak. It is designed for use in various laboratory equipment and processes. The film has a specific sensitivity and characteristics that make it suitable for specific applications, but no further details on its intended use or performance can be provided in an unbiased and factual manner.
Automatically generated - may contain errors
Lab products found in correlation
L7 55, by Beckman Coulter (2 mentions)
Jem 100c transmission electron microscope, by JEOL (2 mentions)
1200exii electron microscope, by Kodak (2 mentions)
Jem 100c, by JEOL (1 mentions)
Jem 1200ex, by JEOL (1 mentions)
Em gp, by Leica (1 mentions)
Uc6 microtome, by Leica (1 mentions)
Jem 1010, by JEOL (1 mentions)
Jem 1200, by JEOL (1 mentions)
Ultrascan 4000, by Ametek (1 mentions)
Whatman, by Cytiva (1 mentions)
Titan electron microscope, by Thermo Fisher Scientific (1 mentions)
626 single tilt cryo transfer system, by Ametek (1 mentions)
Perfection v500 photo flatbed scanner, by Epson (1 mentions)
Tecnai f20 electron microscope, by Thermo Fisher Scientific (1 mentions)
Ammonium molybdate, by Merck Group (1 mentions)
Jem 100cx electron microscope, by Kodak (1 mentions)
17 protocols using so 163 film
1
Phage Particle Morphology Characterization
2
Transmission Electron Microscopy of Phage
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For transmission electron microscopy (TEM) analysis, the high-titer phage preparation underwent CsCl density gradient centrifugation. In this procedure, two mL of the phage preparation was meticulously layered onto a preformed CsCl gradient with specific densities of 1.3 g/mL, 1.4 g/mL, 1.5 g/mL, 1.6 g/mL, and 1.7 g/mL, each occupying 2 mL of the gradient. Ultracentrifugation was carried out using a Beckman Coulter L7-55 ultracentrifuge equipped with an SW 40 Ti rotor. The centrifugation process was conducted at a temperature of 18 °C, with a centrifugal force of 110,961.5 g for a duration of 1.5 h.
Following the density gradient centrifugation, five microliters of the phage suspension was loaded onto 400 mesh carbon-formvar-coated copper grids and negatively stained with 1% uranyl acetate. Phage particles were visualized using a JEM-100C transmission electron microscope (JEOL, Akishima, Japan) with an accelerating voltage of 80 kV. Images were taken on Kodak film SO-163 (Kodak, Cat. #74144, Hatfield, PA, USA) with 45,000× magnification. Phage particle dimensions were measured for 10 phage particles using ImageJ version 1.53e [56 (link)] in relation to the scale bar generated from the microscope.
Following the density gradient centrifugation, five microliters of the phage suspension was loaded onto 400 mesh carbon-formvar-coated copper grids and negatively stained with 1% uranyl acetate. Phage particles were visualized using a JEM-100C transmission electron microscope (JEOL, Akishima, Japan) with an accelerating voltage of 80 kV. Images were taken on Kodak film SO-163 (Kodak, Cat. #74144, Hatfield, PA, USA) with 45,000× magnification. Phage particle dimensions were measured for 10 phage particles using ImageJ version 1.53e [56 (link)] in relation to the scale bar generated from the microscope.
3
Phage Imaging by Transmission Electron Microscopy
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Phage suspension applied onto 400 mesh carbon-formvar coated copper grids was negatively stained with 1% uranyl acetate and subsequently analyzed using a JEM-100С (JEOL, Japan) transmission electron microscope at 80 kV accelerating voltage. Images were taken on Kodak film SO-163 (Kodak, Cat. #74144, Hatfield, PA, USA) with 45000x magnification.
4
Purification and Characterization of Phage
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Three mL of high-titer phage preparation were centrifuged using the preformed CsCl gradient (1.25 g/mL, 1.4 g/mL, 1.5 g/mL and 1.7 g/mL, 2.5 mL each) at 10 °C, 25,000 rpm, for 2.5 h in a Beckman Coulter ultracentrifuge, L7-55, using a SW 41 Ti rotor. Further, ten µL of the concentrated phage suspension (109 PFU/mL) were applied to carbon-coated copper grids (400 mesh) and negatively stained with 1% uranyl acetate. The grids were analyzed using a JEM 1200EX (JEOL, Tokyo Japan) transmission electron microscope at 80 kV accelerating voltage. Images were taken on Kodak film SO-163 (Kodak, Cat. \# 74144, Hatfield, PA, USA). Phage particle dimensions were measured using ImageJ version 1.53e in relation to the scale bar generated by the microscope.
5
Vitrified Dynamin GTPase Imaging
6
Acanthamoeba Infection Ultrastructure Analysis
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Fresh cultures of Acanthamoeba castellanii grown in 10 mL YPG (Yeast extract, peptone, glucose) were infected with 1-week-old 5-µm-filtered suspension of E. lausannensis or W. chondrophila (105 cells.mL−1), as previously reported57 (link). Samples of time-course infection experiments were harvested at 0, 7, 16, and 24 h post infection by centrifuging the infected A. castellanii cultures at 116 × g. Pellets were then fixed with 1 mL of 3% glutaraldehyde for at 4 °C, followed by three washing steps with 100 mM PBS. Cells were further fixed with 1% osmium tetroxide in PBS for 1 h at room temperature. Samples were dehydrated with subsequent increasing ethanol washes (50–100%). Samples were then transferred into propylene oxide and incubated overnight in an epoxy resin (Epon) mixed with 50% propylene oxide58 (link). Thin sections (70-nm thick) were cut with a diamond knife in a Leica UC6 microtome and poststained with periodic acid thiosemicarbazide silver proteinate. All specimens were observed with a Philips CM200 transmission electron microscope operating at 80 kV. Images were recorded on Kodak SO163 film.
7
Visualizing Oligomeric Protein Structures
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Samples were visualized by negative staining in a transmission electron microscope. Aliquots were applied to 400 mesh grids (Maxtaform Cu/Rh HR26) coated with a thin (8 nm) carbon layer and glow-discharged for 20 s. Grids were stained (2 min) with 2% uranyl acetate and air-dried before visualization. Images were acquired using a JEM 1200 (JEOL) electron microscope operated at 100 kV and recorded on Kodak-electron SO-163 film. The diameter of oligomeric structures was measured using ImageJ. Images of CCT-α-syn A53T oligomers were acquired using a JEM 1010 (JEOL) microscope and recorded using a 4K×4K TemCam-F416 (TVIPS) digital camera. Single particles were selected manually and extracted from micrographs using the XMIPP software package64 (link).
8
Ultrastructural Analysis of Lung Tissues
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Lung tissues were fixed in 2.5% glutaraldehyde (Spectrum Chemical, Gardena, CA, USA) overnight at 4°C and washed in PBS (pH 7.4). The tissues were post-fixed in 1% osmium tetroxide (Johnson Matthey Chemicals Ltd., London, UK) and dehydrated in graded ethanol. The tissues were then treated with propylene oxide and embedded in epoxy-resin (Structure Probe Inc., West Chester, PA, USA) embedding medium. The ultrathin tissue sections were stained with uranyl acetate (Resonance Biological Technology Company, Shanghai, China) and lead citrate, and observed using a JEM-1200EX TEM (Hitachi High-Technologies Company, Tokyo, Japan). Images were documented using a SO-163 film (Eastman Kodak Company, Rochester, NY, USA).
9
Phage Visualization Using TEM
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CsCl-purified phage suspensions were applied onto 400-mesh carbon formvar-coated copper grids and stained with uranyl acetate (1% m/v). JEM-100C transmission electron microscope (JEOL, Tokyo, Japan) was used for visualization at an accelerating voltage of 80 kV. Images were recorded on Kodak SO-163 film (Kodak, Cat. No. 74144, Hatfield, PA, USA) at a magnification of 45,000×. Phage particle dimensions were measured for 10 particles using ImageJ version 1.53e [58 ] in relation to the scale bar generated by the microscope software.
10
Cryo-EM Imaging of φEL Protein Complexes
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