Fertilized chicken eggs were ordered from commercial source (EARL Morizeau) and memGFP transgenic chicken eggs were generously provided by Dr. Feifei Song, Dr Adrian Sherman and Dr. Helen Sang (from the Roslin Institute in Edinburgh, Scotland). Eggs were collected at stage X and cultured using a modified version of the EC culture system (Chapman et al., 2001 (link)) until stage 3+ and transferred into bottom glass Petri dish (Mattek inc.) with semi solid albumen/agarose (0.2%) for imaging, with or without drugs: Aphidicolin (30-50μM), Aminopterin (100μM), Jasplakinolide (10μM) and Calyculin A (0.1μM). Embryos were then imaged at 38 degrees using an inverted confocal microscope (Zeiss LSM 700 and LSM880) or a 2-photon Microscope (Zeiss, NLO LSM 7MP) coupled to a Chameleon Ti/Saph laser (Coherent inc.) at 840nm wavelength using 10x, 40X or 63X long distance objectives. The tiling/stitching feature of the Zen software (Zeiss) was used to acquire large embryonic regions (about 1mm2/5000-10000 cells) with a 40x objective. Laser microdissections were performed using a 355nm pulsed laser (35%-50% power), a UGA-42 module from Rapp Optoelectronic coupled to a Zeiss LSM 880 and a 10x objective.
Glass bottom petri dish
Manufactured by MatTek
Sourced in United States
The Glass-bottom Petri dish is a laboratory equipment used for cell culture and microscopy applications. It consists of a standard Petri dish with a transparent glass bottom, allowing for easy observation and imaging of cells or other biological samples under a microscope.
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Lab products found in correlation
Lsm 710, by Zeiss (3 mentions)
Lsm 510 meta, by Zeiss (3 mentions)
Tricaine, by Merck Group (2 mentions)
1 phenyl 2 thiourea, by Merck Group (2 mentions)
Lsm 800 confocal laser scanning microscope, by Zeiss (2 mentions)
Fluoview fv1000 confocal laser scanning microscope, by Olympus (2 mentions)
Epi fluorescence eclipse ti microscope, by Nikon (2 mentions)
Plan fluor 40 1.3 oil objective, by Nikon (2 mentions)
Metafluor software, by Universal Imaging (2 mentions)
Sylgard 184 silicone elastomer kit, by Dow (2 mentions)
Leibovitz s 15 media, by Thermo Fisher Scientific (2 mentions)
Ld lci plan apochromat 25x 0.8 imm corr dic m27, by Zeiss (2 mentions)
Lsm 700 confocal scanning microscope, by Zeiss (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Tcs sp8 confocal laser scanning microscope, by Leica (2 mentions)
Lsm 700, by Zeiss (1 mentions)
Lsm 880, by Zeiss (1 mentions)
Zen software, by Zeiss (1 mentions)
Apotome, by Zeiss (1 mentions)
Vibratome, by Leica (1 mentions)
37 protocols using glass bottom petri dish
1
Imaging Avian Embryogenesis with Confocal Microscopy
2
Chick Embryo Culture and Imaging
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Chick embryos were collected at stage XI and cultured using a modified version of the EC culture system for 5–6 h prior to immunofluorescence processing or up to 15 h for live imaging experiments. Briefly, embryos were collected using paper filter rings and cultured on a semi-solid albumin/agarose nutrient substrate (mixture of albumin, agarose (0.2%), glucose and NaCl) at 37 °C in a humid chamber with or without drugs: DMSO for control embryos and Ritanserin at 20 µM, 50 µM, 100 µM, and 200 µM for treated embryos. For live imaging, embryos collected on filter paper rings were transferred to a bottom glass Petri dish (Mattek inc.) on the same media described above and imaged at 37 °C using an inverted microscope (Zeiss Apotome) with a ×5 objective.
3
Time-lapse Imaging of Cortical Slices
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Cortical slice cultures were prepared and time-lapse imaging was acquired as previously described8 (link)51 (link). About 1 day after in-utero electroporation, embryos were removed and the brain was extracted into ice-cold artificial cerebrospinal fluid containing the following: 125 mM NaCl, 5 mM KCl, 1.25 mM NaH2PO4, 1 mM MgSO4, 2 mM CaCl2, 25 mM NaHCO3, 20 mM glucose pH 7.4 and 310 mOsm l−1. Brains were embedded in 3% low-melting agarose in artificial cerebrospinal fluid and sectioned at 400 mm using a vibratome (Leica Microsystems). Brain slices were transferred on to a slice culture insert (Millicell) in a glass-bottom Petri dish (MatTek Corporation) with culture medium containing (by volume): 66% Basal Medium Eagle (BME), 25% Hanks, 5% fetal bovine serum, 1% N2, 1% penicillin/streptomycin/glutamine (Invitrogen) and 0.66% D -(1)-glucose (Invitrogen). Cultures were maintained in a humidified incubator at 37 °C with constant 5% CO2 supply. Twenty-four hours later, Petri dishes with slice cultures were transferred to an inverted microscope FV1000MPE-IX81ZDC (Olympus) and imaged every 20 min for about 2–3 days. Images were analysed by FluoView (Olympus), Imaris (Bitplane) and Photoshop (Adobe Systems).
4
Immunostaining of Muscle Tissue
5
Zebrafish Fluorescent Vesicle Imaging
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Zebrafish were maintained at standard conditions (Westerfield, 1993 ). Fertilized eggs of wild type embryos (ABC) were injected at the 1-cell stage with 25 ng/µl of respective plasmid DNA. Plasmid h2afx:EGFP-Rab5c (Clark et al., 2011 (link)) was used to label the Rab5c vesicles. At ∼36 hpf embryos with transient expression of fluorescent markers were chosen and mounted in a 35 mm glass bottom Petri dish (0.17 mm, MatTek), using 0.7% low melting agarose (Sigma) containing 0.08% tricaine and 0.003% PTU. A Leica TCS SP5 confocal microscope with a 40× water objective was used for imaging.
6
Oocyte Extraction and Imaging Protocol
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To generate extracts of maturing oocytes, approximately 50 to 100 Tg(actb1:Utr-GFP) oocytes labelling F-actin were incubated in 3.5 ml L15 medium containing 1 μM Lysotracker and 1 μg/ml DHP for 60 to 90 min until GV broke down (oocytes were screened for GVBD under the stereomicroscope). Oocyte extracts were prepared by puncturing oocytes using a spike-micropipette with an inner diameter of 50 to 100 μm (BioMedical Instruments) attached to a syringe allowing for the aspiration of ooplasm-Ygs mixture. The pressure was manually controlled to prevent the aspiration of the medium into the pipette and thus the dilution of the oocyte extract. The ooplasm-Yg mixture was then promptly released into mineral oil (Sigma) inside a glass bottom Petri dish (MatTek) and imaged as described above 15 to 30 min after the extraction procedure had started.
7
Singlet Oxygen Detection in 9L Glioma Cells
8
Imaging Embryonic Pigmentation Inhibition
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Pigmentation of embryos and
larvae was inhibited by 1-phenyl-2-thiourea
(Sigma). The embryos were treated with 100 mg/mL tricaine (Sigma),
mounted in a drop of 1.0–1.5% low melting agarose in egg water
and placed onto a glass-bottom Petri dish (MatTek Corporation, Ashland,
Ma). Fluorescence images were obtained using an LSM800 confocal laser
scanning microscope (Zeiss), an Olympus Fluoview FV1000 confocal laser
scanning microscope (Olympus) or high-end stereoscopic microscopes
(Nikon SMZ25). Three-dimensional-rendered z-stack
images and three-dimensional surface-rendered images and movies were
analyzed and assembled using the IMARIS software (BITPLANE).
larvae was inhibited by 1-phenyl-2-thiourea
(Sigma). The embryos were treated with 100 mg/mL tricaine (Sigma),
mounted in a drop of 1.0–1.5% low melting agarose in egg water
and placed onto a glass-bottom Petri dish (MatTek Corporation, Ashland,
Ma). Fluorescence images were obtained using an LSM800 confocal laser
scanning microscope (Zeiss), an Olympus Fluoview FV1000 confocal laser
scanning microscope (Olympus) or high-end stereoscopic microscopes
(Nikon SMZ25). Three-dimensional-rendered z-stack
images and three-dimensional surface-rendered images and movies were
analyzed and assembled using the IMARIS software (BITPLANE).
9
Confocal Microscopy Imaging of Fish Embryos
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An LSM 700 confocal laser scanning microscope (Zeiss) was used for live and immunofluorescence imaging. Fish embryos and larvae were anaesthetized with a low dose of tricaine, placed in a glass-bottom Petri dish (MatTek) with a layer of 1.2% low melt agarose, and imaged using W N-ACP 20X/0.5 and W Plan-Apochromat 40×/1.0 objective lenses. Immunostained fish sections were imaged using a C Apo 40X/1.1 objective lens. For FITC-Dextran microangiography, fluorescein isothiocyanate (FITC)-Dextran, 2000 kDa (Sigma) was injected into the common cardinal vein and imaged after 10 min. An SMZ 25 stereomicroscope (Nikon) was used for brightfield images of anaesthetized fish.
10
Real-Time Analysis of CD101 Effects on Candida Biofilms
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We used time-lapse microscopy (TLM) to determine the temporal effect of CD101 on the ability of C. albicans to form biofilms. This approach involves the capture of single-frame images at specific intervals in real time, enabling the temporal monitoring of the interactions occurring between the drug and Candida as it forms biofilms. Briefly, following adherence of C. albicans blastospores (yeast forms) to SE for 90 min, the discs were placed in a 35-mm-diameter glass-bottom petri dish (MatTek Corp., Ashland, MA). Next, CD101 (0.25 μg/ml) was added to the petri dish, which was incubated at 37°C to allow for biofilm formation. Phase-contrast images for this interaction were captured in real time over a 16-h period using a Leica DMI 6000 B inverted microscope connected to a Retiga EXi Aqua camera (Q-imaging, Vancouver, British Columbia). To determine the structural changes in the maturing biofilm, the acquisition and analysis of a series of horizontal (x-y) optical sections of the biofilm were performed using Metamorph Imaging software (Molecular Devices, Downingtown, PA). A disc with C. albicans blastospores incubated in growth medium alone with no antifungal was used as a control.
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