HEK293 cells were seeded on 6 well plates and grown to 60% confluency in normal growth medium. Cells were transfected with 2.5 μg of the fully Cre-assembled A-D1-D2 composite plasmid containing mCherry as the GOI (containing siRNA scramble) using lipofectamine 2000 (Invitrogen). 24 hours after transfection, cells were supplemented with indicated amounts of doxycycline (Sigma-Aldrich, Carlsbad, CA USA). 24 h later YFP and mCherry fluorescence was imaged on an inverted DMI-6000 Leica wide-field fluorescent microscope equipped with a Leica DFC 350FX camera with a 20× objective.
Dmi 6000 wide field fluorescent microscope
Manufactured by Leica
The DMI-6000 is a Leica wide-field fluorescent microscope designed for advanced imaging applications. It provides a high-performance optical system for capturing detailed fluorescent images. The microscope is equipped with a range of features and components to enable efficient fluorescence-based visualization and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Doxycycline, by Merck Group (1 mentions)
Dfc350 fx camera, by Leica (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Application suite software, by Leica (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Imagej, by Adobe (1 mentions)
2 protocols using dmi 6000 wide field fluorescent microscope
1
Inducible Expression Assay in HEK293
2
Fluorescence Microscopy of Fixed Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were grown on coverslips and were fixed in −20°C methanol for 10 min. Blocking and antibody dilution buffer was 0.5% BSA (Sigma), 0.1% TritonX100 (Sigma). Fixed cells were visualized with an X63 objective on an inverted DMI-6000 Leica wide-field fluorescent microscope. Sample preparation for TIRF imaging on egg extract was performed as described previously (Petry et al., 2013 (link)). Imaging was performed on a Leica DMI-6000 GSD with a X100 TIRF objective. Pictures were acquired with the Leica Application Suite software. Images were processed with Image J or Photoshop (Adobe) and mounted in figures using Illustrator (Adobe).
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!