For time-lapse imaging, dechorionated embryos were embedded in 1% low-melting agarose within glass-bottom Petri dishes (MatTek Corporation) and submerged in 0.3X Ringer’s solution. For all imaging, the microscope stage was enclosed in a temperature-controlled case, and samples were kept at 28.5°C. For fluorescent images in Fig. 1A and Video 1 , Z-stacks of 4-μm intervals were acquired every 5 s with a 20x/0.75 NA objective on a microscope (Ti-E; Nikon) equipped with a spinning-disk confocal unit (CSU-22; Yokogawa Corporation of America), a charge-coupled device camera (Evolve; Photometrics), and MicroManager software. All other fluorescent images were acquired with a 30x/1.05 NA objective on a microscope (IX83; Olympus) equipped with a spinning-disk confocal unit (CSU-W1; Andor), a scientific complementary metal–oxide–semiconductor (sCMOS) camera (Prime 95b; Teledyne Photometrics), and MicroManager software. For time lapse imaging (Fig. 1B , Fig. 2B , Video 2 , and Video 3 ), Z-stacks of 4 μm intervals were acquired every 10–20 s. For still images in Fig. 4 , 10 hpf embryos were fixed in 4% paraformaldehyde overnight at 4°C, dechorionated, embedded in 1% low-melting agarose within glass-bottom Petri dishes (MatTek Corporation), and submerged in 1X phosphate-buffered saline (PBS); Z-stacks intervals were acquired at 2 μm intervals.
Glass bottom petri dishes
Manufactured by MatTek
Sourced in United States, Morocco
Glass-bottom Petri dishes are a type of laboratory equipment designed for cell culture applications. They feature a transparent glass bottom that allows for easy visualization and imaging of cells under a microscope. The dishes provide a controlled environment for culturing cells and facilitate microscopic observation and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Tricaine, by Merck Group (5 mentions)
Low melting point agarose, by Merck Group (4 mentions)
Lsm 780, by Zeiss (4 mentions)
Lsm 880, by Zeiss (4 mentions)
Dab freebase, by Merck Group (3 mentions)
A1sir confocal microscope, by Nikon (2 mentions)
Clara dr 2199, by Nikon (2 mentions)
Mmessage mmachine sp6 kit, by Thermo Fisher Scientific (2 mentions)
Eclipse ti microscope, by Nikon (2 mentions)
Ultramicrotome, by Leica (2 mentions)
Em grade, by Agar Scientific (2 mentions)
Csu w1, by Oxford Instruments (2 mentions)
Prime 95b, by Teledyne (2 mentions)
Neurobasal a, by Thermo Fisher Scientific (2 mentions)
Sprague dawley strain, by Charles River Laboratories (2 mentions)
Glutamax 1, by Thermo Fisher Scientific (2 mentions)
Papain, by Merck Group (2 mentions)
Lsm 780 nlo confocal microscope, by Zeiss (2 mentions)
Airyscan, by Zeiss (2 mentions)
1 phenyl 2 thiourea ptu, by Merck Group (2 mentions)
61 protocols using glass bottom petri dishes
1
Time-Lapse Imaging of Zebrafish Embryonic Development
2
Tracing Cardiac Progenitors in Zebrafish
3
Tracing Cardiac Progenitors in Zebrafish
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Wild-type and tbx1 mutant embryos carrying the nkx2.5:Kaede transgene were photoconverted using a Nikon A1SiR Confocal Microscope (Nikon Instruments Inc.) and a 20X objective. 16 somite stage embryos were mounted in 0.9% low melting point agarose (Sigma-Aldrich) on 35 mm MatTek glass bottom Petri dishes (MatTek Corporation). Prior to photoconversion, embryos were imaged using a 488 laser to detect green fluorescence. Embryos were then exposed continually to UV light using the 405 mm laser for 3 min. Photoconverted animals were kept in a 28 C incubator until 32 hr post fertilization, anesthetized with 0.04% tricaine (Sigma-Aldrich), and imaged by confocal microscopy. The resulting z stack images were analyzed using FiJi/ImageJ software.
4
Hydrogel Scaffold for Tissue Engineering
5
MDCK Cell Culture Protocol
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Experiments were carried using the MDCK cell line (American Type Culture Collection CCL-34), free from contamination and from young stock. Cells were maintained on several different substrates: collagen coated 35mm diameter glass bottom petri dishes (MatTek Corporation, Ashland MA, product # P35G-1.5-14-C), 60mm plastic petri dishes (Celltreat products, product # 229660, tissue culture treated), and collagen-coated Millicell-CM inserts (inner diameter 25 mm, permeable support area 0.6 cm2; Millipore Corp, Billerica, MA). Cells were grown and maintained at 37°C, 5% CO2. Growth medium consisted of the following (final concentrations): Dulbecco's Modified Eagle Medium w/o glucose and Ham’s F12 at a 1:1 ratio, 5 mM glucose, 15 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), 0.06% NaHCO3, 2 mM L-glutamine, 10% fetal bovine serum (FBS). For differentiation, FBS was reduced to 1%. The amount of medium was restricted so that the apical fluid was thin enough to allow sufficient O2 to diffuse to the monolayer.
6
Live-cell Imaging of Paxillin Dynamics
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Control or Agrin shRNA-tranduced MHCC-LM3 cells were cultured in glass bottom Petri dishes (MatTek Corporation) for 12–16 h before imaging live at 37 °C using the differential interference contrast filter in a confocal microscope (Olympus). For paxillin-GFP imaging, cells were trypsinized and plated as in adhesion assay and imaged 3 h post plating. Each image shown in panels corresponds to a 1-min interval. Data were analysed using the Fluoview software.
7
Visualizing Quorum Sensing Regulation
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Overnight cultures of ΔlasI expressing PlasI-mCherry::attB or PpqsA-mCherry::attB fluorescent fusions were diluted 1:100 in PS:DB and incubated at 37 °C with shaking (250 rpm). Various amounts of 3OC12 HSL was added when cultures reached OD600nm = 0.2. When cultures reached OD600nm = 0.5–0.6, cultures were transferred to glass-bottom petri dishes (Mattek Corporation, Ashland, MA) and incubated for 1 h at 37 °C on an orbital shaker (80 rpm). Planktonic cells were isolated by aspirating the culture media. Surface-attached cells were washed twice in PBS. Samples were covered with 1% agar pad and imaged using fluorescent microscopy.
8
Mitochondrial Imaging in PBMCs
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Resting and activated PBMCs without ALA were seeded on glass bottom Petri dishes (MatTek Corporation, Ashland, MA, USA). Rhodamine 123 (Rh123) (1 µM) was used for mitochondria staining. A Zeiss Axiovert 40CFL microscope (City, Country) with a 100×/1.25 objective was used to image Rh123 using the filter combination of an excitation BP filter 450–490 nm (peak at 475–485 nm) and an emission LP filter >515 nm.
9
Imaging Zebrafish Embryos with Confocal Microscopy
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Zebrafish embryos were treated with phenylthiourea (PTU) at 24hpf, prior to being anesthetized with tricaine and mounted dorsally in 0.9% low melting point agarose on glass-bottom Petri dishes (Mattek) at the desired time. Images were acquired with a Leica SP8 system using PMT or HyD3 detectors and a 20×glycerol immersion objective, HCX PL APO Lambda blue 20×/0.7 argon laser 30%, Diode 405 nm (DPSS 561; Ready Lasers). z-stacks were recorded with 0.57×0.57 × 1.19 μm voxel size. Gain was adjusted to each signal in order to display as little burnt signal as possible.
Stained fixed samples were imaged on a Leica SP8 inverted confocal microscope with 20×glycerol immersion objective and hybrid detectors. For xyz confocal cross-sections, z-stacks were acquired with a 1.194 μm z distance.
Stained fixed samples were imaged on a Leica SP8 inverted confocal microscope with 20×glycerol immersion objective and hybrid detectors. For xyz confocal cross-sections, z-stacks were acquired with a 1.194 μm z distance.
10
CTC Staining of P. aeruginosa Biofilms
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For microscopic analyses, overnight cultures of P. aeruginosa PA14 lasB::Tn were diluted 1:100 in LB, HodC was added at a final concentration of 70 U/ml, and the suspensions were used to inoculate glass bottom petri dishes (MatTek, Ashland, USA). After 24 h of incubation at 37°C under static conditions, planktonic cells were removed from the medium by gently washing with LB medium and the attached viable biofilm cells were stained using 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) for 3 h in the dark as described previously (Li et al., 2013 (link)). Fluorescence microscopy was carried out using an Axioplan 2 imaging system (Carl Zeiss, Oberkochem, Germany) with appropriate filter sets.
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