Following fresh fungal sample collection, cells were immersed in chilled 2.5% glutaraldehyde (Ted Pella Inc.; Redding, CA, USA) in 0.1 M sodium cacodylate buffer (Sigma–Aldrich, St. Louis, MO, USA) and incubated at 4 °C for 1 h before being washed 3 times in 0.1 M sodium cacodylate buffer. Cells then underwent isopropyl alcohol (IPA) dehydration via a series of incremental IPA steps from 50% to 100% (50%, 70%, 80%, 90%, 95%, and 3 times 100%) and stored at 4 °C in 100% IPA. Samples were critically point dried in an Automegasamdri 915B critical point dryer (Tousimis, Rockville, MD, USA). Samples were attached to scanning electron microscopy (SEM) stubs with carbon tape (Ted Pella Inc., Redding, CA, USA), followed by carbon coating with a Leica EM ACE600 Carbon Evaporator (Leica, Wetzlar, Germany) to a thickness of ~12 nm. SEM analysis was performed with an FEI Quanta 200F scanning electron microscope (Thermo Fisher, Waltham, MA, USA).
Carbon tape
Manufactured by Ted Pella
Sourced in United States
Carbon tape is an adhesive-backed tape made of conductive carbon material. It is used to provide an electrical connection or grounding path in various laboratory and scientific applications.
Automatically generated - may contain errors
Lab products found in correlation
S 4700 sem, by Hitachi (3 mentions)
Desk 5 hp, by Denton (3 mentions)
Sem stub, by Ted Pella (3 mentions)
Automegasamdri 915b, by Tousimis (2 mentions)
Leica em ace600 carbon evaporator, by Leica camera (2 mentions)
Sputter coater, by Quorum Technologies (2 mentions)
S 4800, by Hitachi (2 mentions)
Glutaraldehyde, by Ted Pella (1 mentions)
0.1 m sodium cacodylate buffer, by Merck Group (1 mentions)
Fei quanta 200f scanning electron microscope, by Thermo Fisher Scientific (1 mentions)
Originpro 9, by OriginLab (1 mentions)
Orion nanofab helium ion microscope, by Zeiss (1 mentions)
Quanta 200 sem, by Thermo Fisher Scientific (1 mentions)
Smz800n, by Nikon (1 mentions)
Tm 1000, by Hitachi (1 mentions)
Thin coverslip, by Thermo Fisher Scientific (1 mentions)
Sp2 microscope, by Leica camera (1 mentions)
Ultra 55, by Zeiss (1 mentions)
Dlsr camera, by Canon (1 mentions)
Cypher afm, by Oxford Instruments (1 mentions)
23 protocols using carbon tape
1
Fungal Sample Preparation for SEM
2
Fiber Characterization of Electrospun Meshes
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Fiber
morphology and diameter distribution of DEX-loaded electrospun
meshes were determined using SEM. Briefly, samples were mounted on
a metal substrate with carbon tape (Ted Pella, USA) and sputter-coated
(Quorum Technologies, UK) with Au/Pd for 120 s using a 15 mA process
current. Sputter-coated samples were imaged at an accelerating voltage
of 10 kV using a tabletop SEM (Phenom World, Netherlands) fitted with
a backscatter electron detector. ImageJ software (National Institutes
of Health, USA) was used to measure the diameters of at least 100
fibers per sample collected across several SEM micrographs, and histograms
were generated using OriginPro9 software (OriginLab Corporation, USA).
morphology and diameter distribution of DEX-loaded electrospun
meshes were determined using SEM. Briefly, samples were mounted on
a metal substrate with carbon tape (Ted Pella, USA) and sputter-coated
(Quorum Technologies, UK) with Au/Pd for 120 s using a 15 mA process
current. Sputter-coated samples were imaged at an accelerating voltage
of 10 kV using a tabletop SEM (Phenom World, Netherlands) fitted with
a backscatter electron detector. ImageJ software (National Institutes
of Health, USA) was used to measure the diameters of at least 100
fibers per sample collected across several SEM micrographs, and histograms
were generated using OriginPro9 software (OriginLab Corporation, USA).
3
SEM Imaging of PDLLA Nanofiber Scaffolds
4
Helium Ion Microscopy of Silk Fibers
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MAS and MiS fibres were imaged using a Zeiss ORION NanoFab Helium Ion Microscope with SE detection and without metallic coating. Charge compensation was ensured through a low-energy electron beam (flood gun, 600 eV) directed at the sample. Surface sputtering was performed at 25 keV beam energy, with a probe current ranging from 2 to 10 pA. During the cutting the flood gun was continuously on. The silk was cut into a length to fit on top of aluminum specimen mounts (Plano GmbH, Germany) and mounted using carbon tape (Ted Pella, Inc., USA).
5
SEM Analysis of Particle Morphology
6
Scanning Electron Microscopy of Fiber Alignment
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For examination of fiber alignment, each type of EFM construct was attached to an aluminum stub using carbon tape (Ted Pella, Inc., Redding, CA, USA), sputter coated with gold for 30 s (Model 3 Sputter Coater 91000, Pelco, Reading, CA, USA) and imaged using a scanning electron microscope (SEM) (Quanta 200 SEM, FEI Company, Hillsboro, OR, USA). Fast Fourier Transform (FFT) was used to quantitatively validate fiber alignment [29 (link),30 (link)].
7
Preparation of Biological Samples for SEM
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Adult heads were removed, bisected sagittally and fixed overnight in 2% glutaraldehyde + 4% paraformaldehyde in 0.1 M sodium phosphate buffer, pH 7.4. Samples were processed in a critical point dryer, mounted on carbon tape (Ted Pella, Redding, CA 16,084–6) or with colloidal silver (Ted Pella, 16,034) on an SEM stub (Ted Pella, 16,221) and splutter coated with gold-palladium (Ted Pella, 22–2). Images were collected on a JEOL Neoscope Scanning Electron Microscope, and cropped in Photoshop.
8
Characterization of Cellulose Film Morphology
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The morphological changes in the cellulose films were characterized using a stereomicroscope (SMZ800N, Nikon Corporation, Tokyo, Japan) and a scanning electron microscope (SEM, TM-1000, Hitachi, Japan). Cellulose films were placed on the sample stage and visualized with a 1× objective lens of the stereomicroscope. For SEM tests, samples were mounted on the sample stage using carbon tape (Ted Pella Inc., Redding, CA, USA) and visualized at 500× magnifications with an accelerating voltage of 15 kV.
9
Imaging of Birefringent Structures
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Example 4
The present Examples describes imaging.
Birefringence of provided constructs was achieved via polarization microscopy on an inverted microscope. Two linear polarizers (one placed at the light source, and one above provided structures) were used to eliminate direct light and illuminate birefringent material. Images were captured using a Canon DLSR camera. High-resolution images of provided structures were obtained using a Zeiss Ultra55 scanning electron microscope. Structures were removed from their respective molds, and adhered to an SEM stub (Ted Pella) using carbon tape (Ted Pella). To reveal internal morphology of provided structures, a piece of scotch tape was attached to a skin of a structure, which was subsequently peeled so as to reveal the interior. A thin layer (10 nm) of gold was finally sputtered onto substrates before imaging. Confocal images were captured using a Leica SP2 microscope. Samples were inverted within their molds above a thin coverslip (#1, Fisher), and 20 sliced images obtained through their respective depth.
10
SEM Imaging of Sputter-Coated Samples
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Samples were deposited on a standard SEM stub (Ted Pella) that was covered with carbon tape (Ted Pella) and then sputter-coated with Au/Pd (5 nm thickness) using a sputter coater (CCU-010, Safematic). The samples were then imaged using a Verios 460L SEM at 15 kV and 2500 magnifications. Data collection and analysis were carried out with xT Microscope Control software. ImageJ software (National Institutes of Health, http://rsb.info.nih.gov/ij ) was used for quantitative analysis of alignment and porosity.
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