The device was rinsed for a few minutes with a 0.25 M aqueous NaOH solution and water after which the three fluids were pumped through the microfluidic device using syringe pumps (Harvard Apparatus) forming double emulsions. The PTFE tubing and connector were purchased from Achrom, and Dolomite Microfluidics, respectively. The formation of the double emulsion template was observed by an inverted optical microscope (Nikon Eclipse Ti) equipped with a high-speed camera (Fastcam Mini UX50, Photron). The emulsion droplets were collected in a glass vial and pipetted onto a glass coverslip, or directly on a glass coverslip (Menzel-Gläser, 24 × 60 mm, #1) or Petridish (MatTek Corporation, 35 mm Petridish, 20 mm Microwell, #1.5) to be able to directly observe under an optical or confocal microscope. Prior to each experiment, the chip was flushed with NaOH (1 M) in the case of oil-in-water or with silanization solution I (5% in Heptane, Sigma-Aldrich) in the case of water-in-oil emulsions.
Petri dish
Manufactured by MatTek
Sourced in United States
A Petri dish is a shallow, circular dish with a loose-fitting lid, commonly used in laboratories for culturing microorganisms or tissue cells. It provides a controlled environment for the growth and observation of these biological samples.
Automatically generated - may contain errors
4 protocols using petri dish
1
Generating Double Emulsion Templates
2
Cellular Uptake and Localization of RNase A-Based Conjugates
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HeLa cells were planted on a 35 mm × 35 mm Petri dish (MatTek, U.S.A.) with a 10 mm well at a density of 2 × 104 cells per well and cultured overnight at 37 °C under a 5% CO2 environment. After washing with PBS three times, the cells were incubated with 1 mL of fresh culture medium containing AF488-RNase A, sgc8-HDF, AF488-RNase A@HDF or AF488-RNase A@sgc8-HDF each at a concentration of 100 nM, respectively, for 6 h for cellular uptake studies. The cells were then washed with PBS, and the cell nuclei were stained using the Hoechst 33342 fluorescent DNA dye (1 μg mL−1) at 37 °C for 20 min. After washing with PBS, CLSM imaging was performed. For the subcellular localization study, the cells were incubated with 1 mL of culture medium containing AF488-RNase A@sgc8-HDF (100 nM) for 3 and 6 h, respectively. Then, cellular lysosomes and cell nuclei were respectively stained with LysoTracker Red DND-99 (75 nM) and Hoechst 33342 (75 nM), followed by three times of washing with PBS before CLSM imaging.
3
Antibacterial Efficacy of HTC-Based Mats
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The antibacterial activities of the developed HTCs, HTCs-capsaicin, HTCs-AuNP, and HTCs-capsaicin/AuNP mats were estimated and compared with those of difloxacin and chloramphenicol antibiotics, which served as the positive control. Blend suspensions were evaluated to determine their antibacterial activities against Pasteurella multocida, Klebsiella rhinoscleromatis, Staphylococcus pyogenes, and Vibrio vulnificus. Each tested bacterial specie was cultured on a separate Petri dish (MatTek, Ashland, MA, USA) containing Muller–Hinton agar (Qiagen, Mississauga, ON, Canada). Three wells were made in each dish, into which 0.5 mL of each blend suspension was injected. Afterward, the Petri dishes were incubated for 48 h at 37 °C. After the incubation, the inhibition zone was measured and compared with that of the antibiotic materials.
4
Correlative CLEM of HeLa Cells
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Two days prior to infection HeLa LAMP1-GFP cells (1 x 105) were seeded onto a gridded coverslip in a petri dish (MatTek, Ashland, MA). 14 h p.i. 100 ng x ml-1 AHT was added to the cells for induction of reporter plasmid. 16 h p.i. cells were pre-fixed with pre-warmed 2.5% GA in 100 mM phosphate buffer for 15 min at 37°C. After washing the cells thrice with PBS, ROIs were registered and images acquired. Subsequently, further fixation was performed using 2.5% GA in 100 mM phosphate buffer o/n at 4°C. Quenching, osmification and contrasting was performed as described above. Then, the gridded coverslip was removed from the petri dish and was transferred to a glass dish. Afterwards, cells were dehydrated in a cold graded ethanol series, finally rinsing once in anhydrous ethanol and twice in anhydrous acetone at room temperature. Infiltration and flat-embedding were performed in mixes of acetone and EPON812 (Serva). During the removal of the gridded coverslip from the polymerized EPON the engraved coordinates were transferred to the EPON surface and allowed easy relocation by microscopy. ROIs were cut using a scalpel and transferred to an EPON block. Serial 200 nm sections were generated by an ultramicrotome (Leica EM UC7) and collected on formvar-coated copper EM slot grids.
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