For the confocal and widefield fluorescence experiments, the embryos were dechorionated with fine forceps, embedded in 0.3% low melting point agarose in E3, preheated to 42 °C, and gelled on a 35-mm diameter glass bottom dish (ibidi, Gräfelfing, Germany). Once solidified, E3 medium was added. For the bioluminescence experiments, 8-well glass slides (ibidi) were used, and the embryos were embedded in 0.15% low melting point agarose in E3 medium. Embryos over 24 hpf were anaesthetized with 0.03% (1.1 mM) Tricaine (ethyl 3-aminobenzoate methanesulfonate salt, Sigma-Aldrich, Darmstadt, Germany) in E3 for 3 min prior to mounting for confocal microscopy. Embryos were not anesthetized for in vivo Ca2+ measurements.
8 well glass slides
Manufactured by Ibidi
The 8-well glass slides are a laboratory equipment designed for various applications. These slides feature eight separate wells, allowing for multiple samples or reactions to be conducted simultaneously. They are made of high-quality glass, providing a durable and reliable surface for experiments.
Automatically generated - may contain errors
Lab products found in correlation
Tricaine ethyl 3 aminobenzoate methanesulfonate salt, by Merck Group (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
Tcs sp8 sted 3x microscope, by Leica (1 mentions)
3 protocols using 8 well glass slides
1
Zebrafish Embryo Imaging Techniques
2
Enteroendocrine STC-1 Cell Line Exposure
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Enteroendocrine STC-1 (CRL-3254) cell line
was obtained from the American Type Culture Collection (ATCC). Cells
were cultured in Dulbecco’s Modified Eagle’s Medium
(DMEM, Corning) containing 4.5 g/L glucose, 2 mMl -glutamine,
10% (v/v) fetal bovine serum (Life Technologies), penicillin (100
U/mL), and streptomycin (100 μg/mL, Gibco). Cells were incubated
at 37 °C in a humidified atmosphere with 5% CO2. Cells
were serially passaged using 0.25% Trypsin–EDTA (Gibco). Cells
were seeded onto two 8-well glass slides (Ibidi) at a density of 1
× 105 viable cells/well and incubated for 48 h. Then,
the growth medium was replaced with fresh medium supplemented with
either sodium nitrate, sodium nitrite, or vehicle. Cells were incubated
for another 24 h before fixation.
was obtained from the American Type Culture Collection (ATCC). Cells
were cultured in Dulbecco’s Modified Eagle’s Medium
(DMEM, Corning) containing 4.5 g/L glucose, 2 mM
10% (v/v) fetal bovine serum (Life Technologies), penicillin (100
U/mL), and streptomycin (100 μg/mL, Gibco). Cells were incubated
at 37 °C in a humidified atmosphere with 5% CO2. Cells
were serially passaged using 0.25% Trypsin–EDTA (Gibco). Cells
were seeded onto two 8-well glass slides (Ibidi) at a density of 1
× 105 viable cells/well and incubated for 48 h. Then,
the growth medium was replaced with fresh medium supplemented with
either sodium nitrate, sodium nitrite, or vehicle. Cells were incubated
for another 24 h before fixation.
3
Proximity Ligation Assay for Protein Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proximity ligation assay (PLA) was carried out using the Duolink PLA in situ kit (Merck). Only target proteins in close proximity (< 40 nm) generate uorescent signals indicating a successful PLA reaction. PLA was performed according to the manufacturer's instructions with the exception that blocking and dilution of primary antibodies were performed with Sea Block in a step similar to that performed for immunostaining. For PLA, the COV318 cells were xed as described above on 8-well glass slides (Ibidi). Samples were incubated with a combination of anti-CDR1 and primary antibodies against mitochondrial and cytoskeletal proteins over night at 4 ˚C. This was followed by incubation with species-speci c secondary antibodies conjugated to PLA MINUS and PLUS probes. The samples were mounted using Prolong Diamond antifade with DAPI. Imaging was performed with the Leica TCS SP8 STED 3X microscope.
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!