Forty-eight hours after transfection, approximately 5 × 106 cells were scraped, washed twice with PBS and fixed for TEM with 2% glutaraldehyde in phosphate buffer (80 mM Na2HPO4 and 25 mM NaH2PO4, pH 7.4) at 4 °C for 2 h. The fixing solution was removed and the cell pellet was washed with phosphate buffer every 30 min at 4 °C. The cells were post-fixed in 1% osmium tetroxide and embedded in Araldite (Huntsman Advanced Materials). Thin sections (about 1 μm) were stained with toluidine blue and observed by light microscopy in order to select fields. Ultrathin sections were mounted on 200 mesh copper grids and stained with uranyl acetate and lead citrate. The grids containing the sections were observed on a Jeol JEM 1200EX II transmission electron microscope (Servicio Central de Microscopía Electrónica, Universidad Nacional de La Plata) at 80 kV and photographed (ES500W Erlangshen CCD Gatan).
Araldite
Manufactured by Huntsman
Sourced in United States
Araldite is a line of epoxy-based adhesives manufactured by Huntsman Corporation. It is a two-component adhesive system that forms a strong, durable bond when the components are mixed together. Araldite is designed for industrial and professional applications, providing reliable performance in a variety of bonding, sealing, and encapsulation tasks.
Automatically generated - may contain errors
7 protocols using araldite
1
Transmission Electron Microscopy of Transfected Cells
2
Spinal Cord Preparation for Electron Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For electron microscopy, spinal cord specimens were fixed by immersion in 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.4) for 4–6 h at 4°C, postfixed in 1% osmium tetroxide for 2 h, dehydrated in graded ethanol solutions, and embedded in Araldite® (Huntsman, Istanbul, Turkey). Semi-thin sections (1 μm) were then obtained and stained with toluidine blue for subsequent light microphotography and observation. Ultra-thin sections were also obtained and stained with uranyl acetate plus lead citrate for observation under an LEO 906 E transmission electron microscope.
3
Intracellular Uptake of Gold Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The TEM analysis was used to investigate the uptake and intracellular localization of the GNPs inside the cells. The cells were seeded on 6-well plates at a density of 5×105 cells/well and cultured for 24 hours. Then, the cells were exposed to the GNPs (50 nM) for 24 hours. At the predetermined time, the cells were collected, washed three times with PBS, pelleted using centrifugation, and fixed in 2.5% glutaraldehyde for 2 hours. After pellets were washed by PBS, the postfixation with 1% osmium tetroxide was performed.
Finally, the pellets were dehydrated with an ascending series of alcohols before embedding the samples in Araldite® (Huntsman Advanced Materials, The Woodlands, TX, USA). The specimens were cut into ultrathin sections (50~70 nm), which were placed onto copper grids and stained with uranyl acetate and lead citrate. The ultrastructural analysis was performed using a JEM-1011EX instrument (JEOL, Tokyo, Japan).
Finally, the pellets were dehydrated with an ascending series of alcohols before embedding the samples in Araldite® (Huntsman Advanced Materials, The Woodlands, TX, USA). The specimens were cut into ultrathin sections (50~70 nm), which were placed onto copper grids and stained with uranyl acetate and lead citrate. The ultrastructural analysis was performed using a JEM-1011EX instrument (JEOL, Tokyo, Japan).
4
Microfluidic Hanging-Drop Device Assembly
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the final assembly of a device, the sensor modules were inserted into the designated recesses on the microfluidic PDMS chip. Epoxy adhesive (Araldite, Huntsman, Everberg, Belgium) was poured between the microscopy slide hosting the PDMS structures and the PCB to fix and stabilize the device. As an alternative, double-sided adhesive tape could be used to reversibly affix the sensor module.
Prior to each experiment, the assembled device was activated by means of a 30-s oxygen plasma treatment (Diener Electronic GmbH & Co., Ebhausen, Germany), during which a thin PDMS mask with openings at the inlet, outlet, and all drop sites was used to cover all rim structures of the microfluidic hanging-drop device. This selective plasma treatment caused the inherently hydrophobic PDMS inside the circular drop structures and channel locations to enter a hydrophilic state. The covered rim structures remained hydrophobic, ensuring that the liquid would stay contained between them.
Prior to each experiment, the assembled device was activated by means of a 30-s oxygen plasma treatment (Diener Electronic GmbH & Co., Ebhausen, Germany), during which a thin PDMS mask with openings at the inlet, outlet, and all drop sites was used to cover all rim structures of the microfluidic hanging-drop device. This selective plasma treatment caused the inherently hydrophobic PDMS inside the circular drop structures and channel locations to enter a hydrophilic state. The covered rim structures remained hydrophobic, ensuring that the liquid would stay contained between them.
5
Transmission Electron Microscopy of Medulla Oblongata
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Small parts of the medulla oblongata (1 mm×1 mm×2 mm) at the lateral 1.9–2.4 mm level from the center [15 (link)] were taken and immediately immersed in buffered formol glutaraldehyde (pH=7.3). After an hour, the medulla oblongata segments were cut into about 1 mm3 specimens and fixed in buffered formol glutaraldehyde, pH 7.3, for 24 hours at 4°C and routinely osmicated in 1% osmium tetroxide. After dehydration with graded ethanol series, the tissues were embedded in Araldite (Huntsman Advanced Materials, Salt Lake City, UT, USA). Then, TB stain was used to stain the semithin sections as a preliminary step for the selection of the areas needed for the ultrathin examination. Ultrathin sections were stained with lead citrate and uranyl acetate and were viewed under a transmission electron microscope.
6
Hydrogel Stiffness Characterization via AFM
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After their preparation, hydrogels were immersed in PBS and stored at 4 °C until mechanical characterization on the following day. After equilibrating them at RT for 1 h, gels were mounted using CellTak (Thermofisher, Waltham, MA, USA) onto glass object slides (VWR, Radnor, PA, USA). A Nanowizard I or IV (JPK Instruments, Berlin, Germany) was used to probe the gel stiffness. Cantilevers (arrow T1, Nanoworld, Neuchâtel, Switzerland) that had been modified with polystyrene beads of 10 μm diameter (Microparticles GmbH, Berlin, Germany) using epoxy glue (Araldite, Huntsman Corporation, The Woodlands, TX, USA), were calibrated using the thermal noise method implemented in the AFM software (Version 6.1.159, NanoWizard Control Software, JPK Instruments/Bruker, Berlin, Germany). Hydrogels were probed at RT in PBS using a speed of 5 μm/sec and a relative force setpoint ranging from 2.5 to 4.0 nN in order to obtain comparable indentation depths (0.5–1 µm). Force distance curves were processed using the JPK data processing software (Version 6.1.159, NanoWizard Control Software, JPK Instruments/Bruker, Berlin, Germany). Indentation parts of the force curves (approximately 1.5 μm indentation depth) were fitted using the Hertz/Sneddon model for a spherical indenter, assuming a Poisson ratio of 0.5 [42 ,43 (link)].
7
Characterizing Alveolar Macrophage Responses to MWCNT
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AMs were seeded at 0.5×106 cells/well on glass coverslips in 24-well culture plates. Following treatment with 10 μg/mL of MWCNT-0.6 μm or MWCNT-20 μm for 24 hours, AMs were fixed with 2% glutaraldehyde in 0.1 M cacodylate buffer for 30 minutes, followed by three rinses (5 minutes each) with 0.1 M cacodylate buffer. The AMs were then dehydrated with ethanol and sealed onto a SEM stub with Araldite (Huntsman Advanced Materials, Duxford, UK). The samples were then sputter-coated with a thin film of gold and examined under SEM with a Hitachi s4000 (Hitachi, Tokyo, Japan).
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!