Four hours and 80 h zebrafish embryos and larvae exposed to 1 mg mL−1 fruit-based CD during 2 h, were anesthetized with Tricaine 0.04% prior preservation following a sequenced protocol of fixation (with PFA), permeabilization (with MeOH), rehydration (with milliQ water), re-fixation, glycerol impregnation and analysis in 8-well glass bottom μ-slides (Ibidi, Planegg, Germany).
8 well glass bottom μ slides
Manufactured by Ibidi
Sourced in Germany
The 8-well glass bottom μ-slides are a laboratory equipment product designed for cell culture and microscopy applications. They provide a transparent and high-quality glass surface for optimal imaging and analysis.
Automatically generated - may contain errors
4 protocols using 8 well glass bottom μ slides
1
Zebrafish Embryo Preservation Protocol
2
HeLa-JVM Transfection Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa-JVM cells were plated in 8-well glass bottom μ-Slides (ibidi GmbH, Martinsried, Germany) in antibiotic-free DMEM with 10% FBS at a density to achieve approximately 60% confluency per well in 24 h. Wells with siRNA were reverse transfected as described in the following section. After a 24-h incubation, cells were transfected as described above with a pcDNA-EGFP expression plasmid (for siRNA wells) or cotransfected with pcDNA-EGFP and each pcDNA-p38δ expression plasmid. The ratio of DNA to surface area was identical to that used in the 6-well plates. After 24 h, cells were rinsed twice with PBS, then DMEM sans phenol red plus 10% FBS was added to each well. Cells were visualized with a Keyence BioRevo BZ-II 9000 digital microscope fitted with a Nikon PlanApo 4×/0.20 objective lens and 49002 ET-EGFP filter set from Chroma (Bellows Falls, VT). Tiled images covering approximately 70% of each well were stitched with Keyence BZ-II Analyzer software, and total fluorescence in each stitched image was quantified in Fiji software using the Integrated Density function.
3
Quantifying Platelet Adhesion Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Washed human platelets (9 × 106) were seeded on fibronectin-coated (25 μg/ml) 8-well glass bottom μ-slides (Ibidi) and fixed with 4 % formalin after 2, 5, 12, and 30 min incubation at room temperature. Cells were extensively washed with PBS, blocked/permeabilized with 5 % normal donkey serum in PBS containing 0.2 % Triton X-100, and incubated with 1:80 diluted primary Abs in 0.1 % BSA in PBS containing 0.2 % Triton X-100. After extensive washing, platelets were incubated with Alexa Fluor 488/546 conjugated donkey anti-mouse or donkey anti-rabbit Abs (Invitrogen), washed again, and mounted with 50 % glycerol in PBS containing 5 % DABCO (Sigma-Aldrich) as an anti-fading agent. Stained sections were investigated using a confocal laser scanning microscope (LSM 780, Zeiss) equipped with a 63-fold oil immersion objective. Images were acquired and prepared for presentation using the ZEN software (Carl Zeiss, version 2.1, black, 64 bit).
4
Live Cell Imaging of Nuclear Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were plated in 8-well glass bottom μ-slides (ibidi) at least 24 hr prior to imaging. For siRNA experiments, cells were transfected in the chamber slide and hydroxyurea was added at least 4 hr after last transfection, and most often 24 hr after. Live cell imaging was performed using a 20×/0.70 Plan Apo Leica objective or a 40x/1.30 Plan Apo Leica objective (where noted) on an automated Leica DMi8 microscope outfitted with an Andor CSU spinning disk unit equipped with Borealis illumination, an ASI automated stage with Piezo Z-axis top plate, and an Okolab controlled environment chamber (humidified at 37°C with 10% CO2). Long term automated imaging was driven by MetaMorph software (version 7.10.0.119). Images were captured with an Andor iXon Ultra 888 EMCCD camera. For most experiments, cells were imaged every 3 min for 24 hr. For analysis of GFP-BAF and RFP-NLS kinetics and the duration of very short nucleus ruptures, images were acquired every 30 sec for 4 hr (30 sec pass time). For micronucleus experiments cells were imaged every 3 min for 36 hr. These exceptions are noted in the figure legends. Post-processing of image stacks was performed in FIJI. Where noted, RFP-NLS or GFP-NLS signal was gamma adjusted to enhance visibility of cytoplasmic FP-NLS signal.
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!