COCs were obtained from the mated females used for ejaculated sperm collection. Both oviducts were removed and placed in 500 μl medium in a petri dish right before dissecting the uterus of the mated female for sperm recovery. The large cumulus mass was recovered from each oviduct by tearing open the wall of oviduct with a 27g needle. The cumulus mass was gently separated into COCs containing 2–4 oocytes each. A COC was placed on the 14 mm diameter recessed glass insert of a 35 mm petri dish (MatTek Corporation, Ashland, MA; Part Number: P35G-0-14-C) and covered with a 3 mm x 3 mm coverslip supported by four dabs of silicon grease (Corning Inc., Corning, NY) (Fig 2A , arrow). As a negative control, four dabs of silicon grease supporting a coverslip were placed in the same recessed insert (Fig 2A , arrowhead). A total of 50 μl medium was added into the insert and then covered with mineral oil. The observation dish was incubated at 37°C under 5% CO2 in humidified air for at least 1.5 h before addition of sperm.
35 mm petri dish
Manufactured by MatTek
Sourced in United States
The 35 mm petri dish is a circular, shallow, and transparent laboratory vessel used for culturing cells and microorganisms. It has a diameter of 35 millimeters and a height of approximately 10 millimeters. The petri dish provides a sterile and controlled environment for the growth and observation of biological samples.
Automatically generated - may contain errors
6 protocols using 35 mm petri dish
1
Isolation and Preparation of COCs
2
Fission Yeast Strain Preparation and Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
S. pombe strains were grown overnight (14–16 h) on yeast extract media (YE5S) agar plates with supplements (adenine, leucine, uracil, histidine, and arginine) at 25°C. For manipulation and imaging, fresh cells were resuspended in liquid Edinburgh minimal medium with adenine, leucine, uracil, histidine, and arginine (EMM5S) and transferred to the base of a 35-mm Petri dish (MatTek Corporation) coated with 2 µl of 2-mg/ml lectin (Sigma-Aldrich). Free cells were removed by washing with EMM5S, and the Petri dish was filled with 300 µl EMM5S and covered with a coverslip sealed with silicone (GE Bayer Silicones). Cell manipulation and imaging was performed at 25°C in a Bachhoffer chamber (Tempcontrol 37–2 digital; ZEISS).
3
Confocal Imaging of U937 Monocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human U937 monocytes (ATCC Number: CRL-1593.2, Manassas, VA USA) were maintained in RPMI 1640 medium (GIBCO, Grand Island, NY, USA), containing 10% (v/v) heat-inactivated fetal calf serum. For the confocal experiment, all cells were cultured in a 35-mm petri dish (MatTek, Ashland, MA, USA), without any additional intervention. The images of the live cells were captured by confocal microscopy (Olympus FV300, Tokyo, Japan) from 1 to 10 min. The protein concentration was determined using the optical density at 280 and 260 nm (quantified by a GBC cintra 10e, Dandenong, VIC, Australia) and calculated using the following equation: protein concentration (mg ml−1)=1.45 × OD280−0.74 × OD260. CnB (0.25 μM ) was used for confocal experiment.
4
Dual-species biofilm architecture visualization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To comparatively visualize the architecture of single and dual-species biofilms, a cell suspension of 1 × 107 cells/mL of S. mutans was grown in RPMI alone or added to pre-adhered C. albicans in glass-bottom dishes (35 mm petri dish, MatTek Corporation). First, C. albicans (1 × 107 cells/mL) was incubated for 90 min at 37°C; and following incubation, dishes were washed with 10 mM PBS, and GFP-S. mutans was added, and the dishes were incubated statically at 37°C overnight. The following day, dishes were washed with 10 mM PBS and biofilms were stained with concanavalin-A-Alexa Fluor 647 (0.05 mg/mL) for 45 min at 37°C to stain for extracellular matrix. C. albicans hyphae were stained with 0.1% Calcofluor White, which stains cell wall chitin, for 10 min at room temperature. Samples were gently washed with 10 mM PBS and examined by confocal laser scanning microscopy at 40× and 60× magnifications (Nikon Ti2 Spinning Disk). Images were processed using Imaris and Photoshop CS6 software.
5
Transport Chamber for Live-Cell Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A transport chamber made of polyvinyl chloride acetate (PVCA) covered with a glass microscopical cover slip (Fisherbrand®) was built in a 35 mm Petri Dish (MatTek Corporation®). A schematic diagram of the transport chamber is shown in Fig. 4A .
6
Imaging Cytotoxic Effector-Target Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Equal volumes of GFP‐WT FasL‐expressing YT clones and 721.221 B cells were resuspended at a concentration of 0.75–.0.80 × 106 cells mL−1 in serum‐free medium. Prior to this final working concentration, all cells were washed in complete RPMI‐1640 medium. A total volume of 300 µL of target cells was plated on a 35 mm petri dish (MatTek Corporation) for 5 min at 37°C in RPMI‐1640. A 1.5 mL of Imaging Buffer which was prepared in RPMI‐1640 (no phenol red), supplemented with 10% FCS, 25 mM HEPES, Penicillin/Streptomycin, and L‐glutamine, was added to the plated cells. YT cells were spun down in Imaging Buffer for 2 min at RT, resuspended and then added on top of target cells in the petri dish. Cells were fixed at the indicated times and imaged by confocal miscroscopy as described above.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!