Briefly, cells (BxPC-3, 3×104 cells/well and KP-4, 2×104 cells/well) were seeded into CELLview 35-mm glass-bottom cell culture dishes with four compartments for 24 h. These cells were treated with a combination of lapatinib (10 µM) and FTY720 (5 µM) at 37°C for 1 or 4 h. JC-1 dye (5 µM; FUJIFILM Wako Pure Chemical Corporation) was added during the last 30 min at 37°C. Next, the cells were visualized using an LSM 700 confocal laser scanning fluorescence microscope (Zeiss GmbH) equipped with a Plan-Apochromat 63×/1.4 oil DIC (Zeiss GmbH). All images were acquired and processed using ZEN 2012. The object-based fluorescence intensity was measured using ImageJ software.
Lsm 700 confocal laser scanning fluorescence microscope
Manufactured by Zeiss
The Zeiss LSM 700 is a confocal laser scanning fluorescence microscope. It is designed to capture high-resolution, three-dimensional images of fluorescently labeled samples.
Automatically generated - may contain errors
3 protocols using lsm 700 confocal laser scanning fluorescence microscope
1
Evaluating Mitochondrial Dysfunction in Cancer Cells
2
Calcium Imaging of Cell Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Briefly, cells (BxPC-3, 3×104 cells/well and KP-4, 2×104 cells/well) were seeded into CELLview 35-mm glass-bottom cell culture dishes with four compartments. Then, 2 days after seeding, the culture medium was replaced with fresh medium containing 5 µM Fluo-8-AM (AAT Bioquest, Inc.) and cultured at 37°C for 1 h. After washing, the cells were treated with a combination of lapatinib (10 µM) and FTY720 (5 µM) at 37°C for 1 h. Cells were then visualized using an LSM 700 confocal laser scanning fluorescence microscope (Zeiss GmbH) equipped with a Plan-Apochromat 40×/1.4 oil DIC (Zeiss GmbH). All images were acquired and processed using ZEN 2012. The object-based fluorescence intensity was measured using ImageJ software.
3
Microvilli Formation by Overexpression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To grow microvilli on the apical surface, 5 × 105 MDCKmCherry‐ESPIN1 cells were grown on 0.45‐μm polyester filter insert (12‐mm‐diameter Transwell, Corning) for 48 h to obtain complete confluency. Then, plasmids were transfected into cells using Lipofectamine LTX (Invitrogen). Twenty‐four hours after transfection, cells were fixed with 4% paraformaldehyde (PFA) in 0.1 M PBS (pH 7.4). HeLa cells were seeded in 35‐mm glass‐bottomed dishes (MatTek), 48 h before transfection, and transfected using FuGENE 6. Thirty‐two hours after transfection of plasmids, the cells were fixed using 4% PFA solution. After permeabilization with PBS containing 0.3% Triton X‐100 (PBS‐0.3T), fixed cells were stained with Alexa568‐conjugated phalloidin with/without DAPI for 1 h at 23°C. Fluorescent images (2D and 3D) were obtained using a LSM700 confocal laser‐scanning fluorescence microscope (Zeiss). For statistical analysis of the effect of DIA1 proteins on microvilli, the longest microvilli of HeLa cell from 3D reconstructed image were measured (n = 10 from five independent experiments).
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!