EB formation was performed as described previously (Takahashi et al., 2007 (link)). Briefly, PSCs were harvested by disassociating the cells with Accutase (STEMCELL Technology, Canada, 05946) and transferred into Nunc Non‐Treated Multidishes (ThermoFisher Scientific, USA, 150239). The cells were incubated in ncEpic medium with Blebbistatin (Nuwacell Biotechnologies Co., Ltd, China, RP01008) for 6 h. Then the medium was replaced with E6 medium (STEMCELL Technology, Canada, 05946), and the E6 medium was changed every other day. After 8 days, the EBs were transferred to a 0.1% Gelatin (Sigma‐Aldrich, USA, G7041)‐coated plate and cultured with DMEM/F12 medium (Corning, USA) containing 10% FBS for 6 days. The medium was changed every other day.
Nunc non treated multidishes
Manufactured by Thermo Fisher Scientific
Sourced in United States, Czechia, United Kingdom
Nunc Non-Treated Multidishes are a line of cell culture vessels designed for various cell-based applications. These multidishes provide a reliable and consistent platform for cell growth and experimental studies.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using nunc non treated multidishes
1
Embryoid Body Formation from PSCs
2
Cryosectioning and Preparation of Lung Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A six-well plate (Nunc Non-Treated Multidishes, Thermo Fisher Scientific, Brno, Czech Republic) was filled with 2 of fixative solution (3% glutaraldehyde, 0.1 M cacodylate buffer, pH 7.4) and frozen in the freezer at
. Before cryosectioning, the six-well plate and deep-frozen lung lobes were conditioned in a cryo-chamber at
. Then, 25 –100 -thick frozen sections of the lung tissue were carefully placed onto the solid surface of the fixative solution with one section per well (Figure 1 , green path). The lid-covered plate was transferred into the refrigerator and maintained at 4 until all the fixative solution completely melted, usually overnight. During the thawing process, the sections continued to float on the fixative solution and were gently fixed by glutaraldehyde diffusion. Subsequently, the sections were transferred onto the cooled cacodylate buffer level and allowed to float for 20 min three times. Washed sections were mounted onto wet coverslips and wrapped in fine copper wire mesh for easier handling. Then, the whole mounts were dehydrated in a graded ethanol series (25, 50, 75, 90, 96, 100, 100%, 20 min each) and dried in a critical point dryer. Dried sections were mounted onto aluminum specimen stubs using ultrasmooth carbon discs (SPI Supplies, Structure Probe, Inc., West Chester, PA, USA).
. Before cryosectioning, the six-well plate and deep-frozen lung lobes were conditioned in a cryo-chamber at
. Then, 25 –100 -thick frozen sections of the lung tissue were carefully placed onto the solid surface of the fixative solution with one section per well (
3
Comprehensive B-cell Niche Modeling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Isolated CD19+ B-cells or sorted subsets were cultured in a 1:5 ratio with supporting cells (CD19+ B-cells:CD40LCs and CD19+ B-cells:LSCs) or 1:2.5:2.5 ratio (CD19+ B-cells:CD40LCs:LSCs) in the presence of an indicated cytokine and molecule mix (Table 1 ). In 2D, supporting cells formed a confluent layer at the bottom of a tissue culture treated 48-well plate (Cellstar Cell Culture Multiwell Plates, Greiner Bio-One), on top of which the CD19+ B-cells were cultured. In 3D, CD19+ B-cells, CD40LCs and/or LSCs were homogenously incorporated throughout an indicated hydrogel composition (Table 2 ) by resuspension, 20 μL gels were casted in the center of a non-tissue culture treated 48-well plate (Nunc Non-Treated Multidishes, ThermoFisher) to prevent cellular outgrowth from the 3D hydrogel. All co-cultures were cultured for 4, 7, 11 and/or 14 days in an humidified incubator at 37 °C at 5% CO2. The number of 30.000 cells per well and added medium volumes were kept equal in both 2D versus 3D. On each time point, all cells from each individual culture well were collected and measured in its entirety, resulting in an absolute total cells count that could be compared over time per condition. Medium was refreshed twice a week, adding 250 μL per well, the supernatant of each time point was stored at −20°C for further analysis.
4
Biofilm Formation on p-PVC Coupons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A 1.8 mL volume of sterile broth was added to each well of a sterile 12-well plate (Nunc Non-Treated Multidishes, Thermo Fisher Scientific, UK) along with a sterilised p-PVC coupon, 11 mm in diameter, test side of the surface face up. The reverse end of a sterile swab was used to push the coupon to the bottom of the well. The 12 well plates were left at room temperature overnight to check for any contaminant growth and to check that all coupons were fully sterilised. An overnight culture was prepared and diluted to obtain 1.0 x 10 8 CFU mL -1 . Two hundred microliters of the bacterial suspension was added to the appropriate wells to give a working concentration of 1.0 x 10 7 CFU mL -1 . The 12 well plates were parafilmed to prevent the media from evaporating from the plates. The plates were incubated at 22 °C for either 5 or 7 days with gentle shaking (68 rpm). Following incubation, all the media in each well was removed by pipetting, leaving just the coupon behind and the wells containing the coupons were washed by adding 2 mL of sterile dH2O to each well; carefully against the side of the well so as not to disturb the biofilm formed on each coupon. The liquid was gently swirled around the wells manually for 5 s before pipetting out the liquid to remove any unbound planktonic cells. The 12 well plates and coupons were air dried in a class II cabinet for 1 h.
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!