Four strains (two strong biofilm producers, one biofilm-negative producer, and ATCC19606) were selected for biofilm visualization by CLSM (Leica, Germany). In brief, strains were cultured in LB broth for 18 h and diluted to 0.5 McFarland standard. One hundred microlitre suspension and 2 mL fresh LB broth were added in the petri dishes (FengRun, Beijing, China). After incubation at 37 °C for 24 h, the petri dishes were washed three times with PBS and fixed with 2 mL of 2.5% glutaraldehyde for 1.5 h. Then, the petri dishes were washed three times with PBS again and 200 μL fluorescein isothiocyanate-concanavalin A (FITC-ConA) (Sigma, Germany) were added and stored at 4 °C for 30 min. Two hundred microlitre propidium iodide (PI) (Sigma, Germany) were added and stored at 4 °C for 15 min after washing three times with PBS. Finally, the dishes were washed with PBS and sealed with 40% glycerol. FITC-conA binds to EPS of biofilm and reflects green fluorescence at the laser wavelength of 488 nm.19 (link) PI binds to bacterial DNA and emits red fluorescence at 543 nm.20 (link) If both lasers appear at the same time, an orange fluorescence will be shown.21 (link)
Petri dishes
Manufactured by Merck Group
Sourced in United States, United Kingdom, Germany
Petri dishes are circular, shallow containers made of glass or plastic, commonly used in microbiology laboratories. They serve as a controlled environment for culturing microorganisms, allowing for the observation and study of their growth and behavior.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Hank s balanced salt solution, by Thermo Fisher Scientific (3 mentions)
Tryple express, by Thermo Fisher Scientific (3 mentions)
96 well plate, by Merck Group (2 mentions)
1640 medium, by Thermo Fisher Scientific (2 mentions)
Conical tube, by Merck Group (2 mentions)
Polystyrene co maleic anhydride, by Thermo Fisher Scientific (2 mentions)
N 3 dimethylaminopropyl n ethylcarbodiimide hydrochloride edac, by Thermo Fisher Scientific (2 mentions)
Antibiotic solution of penicillin streptomycin, by Merck Group (2 mentions)
L glutamine, by Merck Group (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Low serum growth supplement, by Thermo Fisher Scientific (1 mentions)
Circular glass coverslips, by Thermo Fisher Scientific (1 mentions)
Rat tail collagen type 1, by BD (1 mentions)
Hepes, by Merck Group (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
96 well microtiter plate, by Merck Group (1 mentions)
Static incubator, by Memmert (1 mentions)
Nutrient broth, by Merck Group (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
20 protocols using petri dishes
1
Biofilm Visualization by CLSM
2
Functionalization of Endothelial and Tumor Cells
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Endothelial cell (EC) and HT29 tumour cell monolayers were grown on circular glass coverslips before functionalization with particles55 56 (link). Then, 40 mm diameter circular glass coverslips (Thermo Scientific) were plasma treated (Harrick Plasma Cleaner) for 2 min and subsequently incubated in 1% polyethylenemine at room temperature (RT) for 10 min. Coverslips were then washed in deionized water three times and treated with 0.1% glutaraldehyde (Sigma-Aldrich) in PBS at RT for 30 min. Coverslips were washed in deionized water three times, dried and treated with 0.1 mg ml−1 type I rat-tail collagen (Becton Dickinson) in HEPES (pH 8.0; Sigma-Aldrich) for 2 h at 4 °C. Coverslips were placed in Petri dishes (60 mm × 15 mm; Sigma-Aldrich), washed three times in PBS and briefly sterilized via ultraviolet exposure for 15 min. HUVECs were plated on coverslips, at a density of 500,000 cells per coverslip, in Medium 200 supplemented with low-serum growth supplement (Cascade Biologics), 5% FBS (Invitrogen) and 100 U ml−1 PenStrep. HT29 cells were plated at a density of 500,00 cells per coverslip in McCoy's 5A modified medium supplemented with 10% (vol/vol) FBS and 1% (vol/vol) Penstrep. Both cell types were cultured for ∼4 days on coverslips before functionalization with particles.
3
Cell Proliferation Kinetics Monitoring
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The proliferation kinetics and size of the cells were monitored by use of a CASY® Cell Counter and Analyzer System (TTC-2EA-1087, Schärfe System GmbH, Reutlingen, Germany) which was used as described by the manufacturer (Schärfe-System GmbH, Reutlingen, Germany). Cell lines were grown in Petri dishes (Ø 6 cm, Sigma-Aldrich, St Lois, Missouri, USA) in 5.0 ml medium; ca. 5 × 105 cells were seeded in each dish at the start of the experiments. After 24 h intervals, the cells were detached with trypsin–EDTA (Szabo-Scandic, Vienna, Austria) and 50 µl of these suspensions were transferred to CASY-cups (OLS OMNI Life Science GmbH & Co. KG, Bremen, Germany). For each experimental point, three plates were evaluated. On the basis of the results, means and standard deviations (SDs) were calculated. Nonlinear fits with exponential growth equations (least square fits) were calculated to determine the doubling times (http://www.graphpad.com/guides/prism/5/user-guide/prism5help.html?reg_exponential_growth.htm ).
4
Bacterial Growth Media Preparation
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All chemicals and dehydrated bacteriological media were provided by (Sigma-Aldrich, United Kingdom). Petri dishes, 96-well microtiter plates and different reagents were purchased from (Sigma-Aldrich, United Kingdom). Tips, microcentrifuge tubes, solutions and growth media were sterilized in an autoclave at 121°C for 30 min.
5
Trichoderma Inhibition of Plant Fungi
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Nucleation dynamic assay (single or dual) was used to study the ability of the Trichoderma strains to inhibit the growth of plant-associated and plant-pathogenic fungi in 90 mm × 15 mm Petri dishes (Sigma-Aldrich). PDA plates were inoculated with 7 mm diameter mycelial plugs of the active growth stage Trichoderma strains, placed 10 mm from the edge of the plate. Mycelial plugs, 7 mm of each tested fungus, were placed separately, opposite to Trichoderma stains, 25 mm away from each of Trichoderma discs. Control plates were single-inoculated with the Trichoderma strains, while Trichoderma strains were also tested against each other in a dual assay. The plates were incubated in a static incubator (Memmert GmbH + Co. KGat) at 27 °C for 6 days (i.e., 144 h). All cultures were performed in duplicate.
6
Rearing Anopheles and Aedes Mosquitoes
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Anopheles gambiae mosquitoes were the ‘G3’ strain (MRA-112) obtained from the Malaria Research and Reference Resource Center (MR4, BEI Resources, Manassas VA USA). Aedes aegypti were the ‘New Orleans’ strain (NR-49160), also obtained from the MR4 and were in the F16-F18 generations during experiments. A standard rearing water was made of 0.3 g of pond salts (API, McLean, VA USA) per liter of type II purified water. Anopheles gambiae eggs were collected, held overnight on damp filter paper and placed in trays on the day of hatching. Aedes aegypti eggs were dried for embryonation for 4 days after collection before storage in plastic bags in a plastic box. Within 1 month, they were hatched by placing egg papers in water under vacuum for 30 min. Hatching embryos of both species were placed in 500 ml of water containing 5 intact Koi pellets for one day before counting 80 larvae into 150 ml polystyrene Petri dishes (Item no. Z717231, Sigma-Aldrich, St. Louis, MO USA).
7
Isolation and Characterization of Copepod-Associated Bacteria
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Copepod eggs were washed three times with sterile distilled water and then mixed for 3 min using a homogenizer Ultra-turrax Ika-T25D model (Imlab, Lille, France) in 9 mL of SSB (1 g tryptone, 8.5 g sodium chloride dissolved in 1 L of distilled water), serially diluted in 0.9% (v/v) SSB from 10-1 to 10-6, plated on nutrient agar in Petri dishes (Sigma–Aldrich, St. Quentin Fallavier, France) using the spread plate method and incubated at 25°C for 48 h. Morphological characteristics of isolates were determined using visual assessment of bacterial colonies on nutrient agar plates along with microscopic examination according to the standard procedures described in Bergey’s manual of systematic bacteriology. Gram staining and the catalase test (catalase production) were carried out. Morphologically different colonies were selected from the agar plates, streaked on fresh nutrient agar plates and incubated at 25°C for 48 h. Finally, bacterial isolates were inoculated into nutrient broth (Merck, Germany) and stored in 30% glycerol at -80°C for further analysis.
8
Zebrafish Embryo Toxicity Screening
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The zebrafish (Danio rerio) individuals came from the Experimental Medicine Centre, Medical University in Lublin, Poland. All the experiments on embryos and non-feeding larvae were carried out at this Experimental Medicine Centre.
As soon as possible after fertilisation, the eggs were transferred into Petri dishes (Sigma-Aldrich, Saint Louis, MO, USA) containing a fresh E3 medium. After washing and removing debris, only viable fertilised eggs with a round chorion were placed into 24-well sterile cell culture plates (Costar, Corning Inc., USA) (five embryos per well). Each well contained 2 ml of the test solution at a respective concentration or the E3 medium alone. Twenty embryos were used for each concentration of compound/pemetrexed. An equal number of embryos untreated with any compound/pemetrexed was employed as the control group. Throughout the experiment, all the covered plates with embryos/larvae were kept in the incubator at static conditions at 26 ± 1 °C up to 120 h after fertilisation.
As soon as possible after fertilisation, the eggs were transferred into Petri dishes (Sigma-Aldrich, Saint Louis, MO, USA) containing a fresh E3 medium. After washing and removing debris, only viable fertilised eggs with a round chorion were placed into 24-well sterile cell culture plates (Costar, Corning Inc., USA) (five embryos per well). Each well contained 2 ml of the test solution at a respective concentration or the E3 medium alone. Twenty embryos were used for each concentration of compound/pemetrexed. An equal number of embryos untreated with any compound/pemetrexed was employed as the control group. Throughout the experiment, all the covered plates with embryos/larvae were kept in the incubator at static conditions at 26 ± 1 °C up to 120 h after fertilisation.
9
Cell Proliferation Measurement via CASY Analyzer
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The proliferation of the cells was studied by use of a CASY® Cell Counter and Analyzer System (TTC‐2EA‐1087, Schärfe System GmbH, Reutlingen, Germany) which was used as described by the manufacturer (Schärfe‐System GmbH). Cell lines were grown in Petri dishes (Ø 6 cm, Sigma‐Aldrich) in 5.0 mL medium; ca. 5 × 105 cells were seeded in each dish at the start of the experiments. After 24 hr intervals, the cells were detached with trypsin–EDTA (Szabo‐Scandic, Vienna, Austria) and 50 μL of the suspensions were transferred to CASY‐cups (OLS OMNI Life Science GmbH & Co. KG, Bremen, Germany). For each experimental point, three plates were evaluated. On the basis of the results, means and standard deviations (SDs) were calculated. Nonlinear fits with exponential growth equations (least square fits) were calculated to determine the doubling time.
10
Antibacterial Activity of PCL Polymer
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PCL with molecular weight (M.W.) 80,000 g·mol−1, methanol, chloroform, GEN, Mueller Hinton Agar, 12-well plates, 96-well plates, and Petri dishes were acquired from Sigma-Aldrich. Dimethyl sulfoxide (DMSO) was obtained from Honeywell, Fluka, Seelze, Germany. The bacterial strains E. coli ATCC 25922, S. aureus ATCC, B. cereus ATCC 11778, and P. aeruginosa ATCC 10,145 were obtained from the American Type Culture Collection (ATCC). The culture medium DMEM/F12 and the culture medium supplemented with 10% fetal calf serum, streptomycin, and penicillin were acquired from Gibco LifeTechnologies, Paisley, UK. All commercial materials were used without further purification.
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