Approximately, 1 × 103 cells in 0.3% agarose were seeded in a 96-well plate onto a layer of 0.6% agarose. Cells were grown in the present of LB100, carboplatin, or LB100/carboplatin for 3 weeks to observe colony formation. The colonies were fixed in 4% formaldehyde and stained with crystal violet. Z-stacks of tiled bright-field (BF) images were taken using a 5× objective with a step size of 200 μm on a Zeiss Observer 7 inverted microscope (Carl Zeiss). Using Zen Blue v2.5 (Carl Zeiss Microimaging), stacks were processed by first stitching a reference slice, and then the Extended Depth of Focus module, with default settings, was used to compress the Z-stack information into a single image. Manual counting was conducted on the resulting tiled image using the points tool, and summary measurements generated, in QuPath 0.1.3 (31 (link)).
Observer 7 inverted microscope
Manufactured by Zeiss
The Zeiss Observer 7 is an inverted microscope designed for a variety of laboratory applications. It features a stable and ergonomic design, with an adjustable eyepiece and stage to accommodate different user needs. The microscope is equipped with high-quality optics for clear and detailed imaging. The core function of the Observer 7 is to provide a versatile and reliable platform for various microscopy techniques.
Automatically generated - may contain errors
Lab products found in correlation
Zen blue v2, by Zeiss (1 mentions)
Synergy h1 hybrid multi mode microplate reader, by Agilent Technologies (1 mentions)
Dcfda h2dcfda cellular ros assay kit, by Abcam (1 mentions)
Cck 8, by Dojindo Laboratories (1 mentions)
Live dead viability cytotoxicity kit, by Thermo Fisher Scientific (1 mentions)
2 protocols using observer 7 inverted microscope
1
3D Colony Formation Assay with Microscopy
2
Intracellular ROS Detection and Cellular Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The intracellular ROS was determined by using a DCFDA/H2DCFDA-Cellular ROS Assay Kit (Abcam#113851) (2′,7′-dichlorofluorescin diacetate) according to the manufactory’s protocol. ARPE cells were transfected with indicated plasmid and then treated with DMSO or NAC (1 mM) or MitoQ (1 μM) for 6 h. The viable cell amount was determined using CCK-8 (Dojindo Molecular Technologies, #CK04). The resulting signal was recorded using Synergy H1 Hybrid Multi-Mode Microplate Reader (BioTek Instruments, USA). Cell viability was determined using LIVE/DEAD™ Viability/Cytotoxicity Kit (Invitrogen, #L3224) and cells were observed under a ZEISS Observer7 inverted microscope (Carl Zeiss). Cells were transfected with FLAG or FLAG-STING plasmids. After 24 h, cells were treated with or without H2O2 (1.8 mM: Fig. 6G , 0.6 mM: Fig. 6H .) for 2 h and then recovered for 12 h before analysis. For quantification, dead cells were normalized to the fluorescence intensity of live cells.
NAC (1 mM) was added 2 h before H2O2 treatment and then maintained throughout the experiment.
NAC (1 mM) was added 2 h before H2O2 treatment and then maintained throughout the experiment.
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!