WM266 and HeLa cells were seeded on a 35 mm diameter Fluorodish Cell Culture Dish (World Precision Instruments, Inc., Florida) at the concentration of about 5×104 cells/mL. 24 h after seeding, cells were incubated with FITC-RGDechi-hCit and FITC-RGD scrambled at the final concentration of 50 µM in HBSS buffer for 30 min at 37°C. Cells were rinsed twice with PBS to remove peptide excess and fixed with 4% paraformaldehyde at room temperature. Samples were then observed with a confocal microscope (Leica SP5) with a 63× oil immersion objective with a 488 nm wavelength laser line and transmitted light.
Fluorodish cell culture dish
Manufactured by World Precision Instruments
Sourced in Uruguay, Germany
The Fluorodish Cell Culture Dish is a specialized laboratory equipment designed for cell culture applications. It features a clear bottom that allows for optical imaging and analysis of cells. The dish is made of high-quality materials to provide a sterile and controlled environment for cell growth and experimentation.
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Lab products found in correlation
Mem non essential amino acids solution, by Thermo Fisher Scientific (1 mentions)
Gentamycin, by Thermo Fisher Scientific (1 mentions)
Nunc lab tek 2 chambered coverglass, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Sp5 confocal microscope, by Leica (1 mentions)
Volocity 3d image analysis software, by PerkinElmer (1 mentions)
Wavefx spinning disk confocal system, by Quorum Technologies (1 mentions)
Dm6000b inverted microscope, by Leica (1 mentions)
F9423, by Merck Group (1 mentions)
7 protocols using fluorodish cell culture dish
1
Cellular Uptake of FITC-RGD Peptides
2
Cell Culture Protocols for C2C12 and HeLa Cells
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C2C12 cells were cultured at 37 °C with 5% CO2 using DMEM high glucose (4.5 g l−1 of glucose), containing 10% fetal bovine serum, 2% HEPES 1 M, 1% MEM Non-Essential Amino Acids Solution (100 × ) and 40 μg ml−1 of Gentamycin (all products were purchased from Life Technologies Corporation). Cells were plated in 35 mm Fluorodish Cell Culture Dish (World Precision Instruments) at roughly 50,000 cells per plate on the day before fixation.
HeLa cells were cultured at 37 °C with 5% CO2 using Minimum Essential Medium Eagle with Earle’s salts,L -glutamine, sodium bicarbonate complemented with 10% fetal bovine serum, 1 × penicillin-streptomycin, 1 × GlutaMAX, 1 × MEM Non-Essential Amino Acids Solution (all products were purchased from Life Technologies). Cells were plated in 4-well Nunc Lab-Tek II Chambered Coverglass (Thermo Fisher Scientific).
HeLa cells were cultured at 37 °C with 5% CO2 using Minimum Essential Medium Eagle with Earle’s salts,
3
Aplysia Bag Cell Neuron Culture
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Aplysia bag cell neurons were collected from the abdominal ganglion as described previously (Lee et al., 2008 ), and cultured on a glass bottom dish (Fluorodish Cell Culture Dish, World Precision Instruments, Sarasota, FL) coated with 20 ug/ml PLL and immersed in L15 medium supplemented with artificial sea water (ASW; 400 mM NaCl; 9 mM CaCl2; 27 mM MgSO4; 28 mM MgCl2; 4 mM l -glutamine; 50 mg/ml gentamicin; 5 mM HEPES, pH 7.9). After plating, the cells were kept at 14°C and typically used for experiments 1 d after plating.
4
Enamel Surface Preparation for SCFS
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Enamel samples (3 × 4 × 1 mm) were prepared from the vestibular surfaces of bovine incisor teeth. The surfaces were progressively polished by wet grinding with up to 4000 grit (Buehler, Düsseldorf, Germany) and purified from impurities by incubation in 3% NaOCl for 3 min, ultrasonication with distilled water for 10 min, incubation in 70% isopropyl alcohol for 15 min and final incubation in distilled water for at least 12 h. For SCFS measurement, the enamel sample was fixed on a FluoroDish Cell Culture Dish (World Precision Instruments GmbH, Friedberg, Germany) with instant glue (Pattex, Düsseldorf, Germany) and immediately covered with 20 µL PBS.
5
Probing Charge-Dependent Particle Interactions
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Carboxyl-terminated fluorescent Nile red polystyrene particles (1% w/v, 0.84 μm, Spherotech) were diluted by 1:50 in DI water. Amino-terminated fluorescent yellow particles (1% w/v, 0.81 μm, Spherotech) were, firstly, diluted by 1:50 in 0.1% poly-l -lysine (PLL), followed by incubation for 1 hour. Then, the solution was centrifuged at 5000 rpm for 5 min and resuspended with the same volume of DI water. Zeta potential measurements on a Zetasizer (Malvern Nano ZS) confirm that the carboxyl fluorescent Nile red particles and PLL-treated amino fluorescent yellow particles have negative and positive charges on the surfaces, respectively. The particles were then mixed with intrinsic nanowires, and 10 μl of the mixture was added on a FluoroDish Cell Culture Dish (World Precision Instruments) and covered with a coverslip. The single-particle manipulation was performed using the same Leica SP5 confocal microscope. In a typical experiment, a laser pulse (1 ms, 592 nm) was delivered to spots of interest in the middle of an imaging time series.
6
Imaging Zebrafish Embryo Development
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Embryos were collected for 30 min, then dechorionated using 5% bleach for 1 min and rinsed with water. Embryos were then aligned in a Fluorodish cell culture dish (World Precision Instruments) and covered with Halocarbon oil 400. Live imaging was performed using a WaveFX spinning disk confocal system (Quorum) and a DM6000B inverted microscope (Leica). Images were captured and processed using Volocity 3D image analysis software (PerkinElmer).
7
Holo-tomographic Imaging of Live Cells
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Holo-tomographic video imaging was performed on a NanoLive (Switzerland) 3D Cell Explorer fluo (AXT Pty Ltd., Warriewood, NSW) equipped with a NanoLive live cell incubator (AXT Pty Ltd). 1 × 104 cells were seeded into a FluoroDish cell culture dish 35 mm, 23 mm well (World Precision Instruments, FD35) and maintained in phenol red free DMEM medium (Sigma-Aldrich, D1145) supplemented with 10% fetal bovine calf serum (Sigma-Aldrich, F9423), 2 mM glutamine (Sigma-Aldrich, G7513) and 1% penicillin-streptomycin for 48 h. Immediately prior to imaging the medium was removed and replaced with 400 μL of the same medium, followed by transfer to the live cell incubator chamber of the 3D Cell Explorer. Cells were incubated at 37 °C, 5% CO2 and 100% humidity for the duration of the time-lapse. Three dimensional holo-tomographic images were captured every 20 s for the duration of the time-lapse using the Nanolive STEVE software. For File S7 the center plane of each 96 slice stack was exported after capture using the built in STEVE export wizard as an .avi movie file. These files were exported at 5 frames per second (100x actual speed) to visualize cellular dynamics.
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