For live-cell TIRF microscopy, transfected cells were visualized on a TIRF microscope (Roper Technologies) equipped with an ILas2 double laser illuminator (Roper Technologies), a CFI Apo TIRF 100× (1.49-NA) objective (Nikon), and an Evolve512 delta EMCCD camera (Photometrics). Image acquisition was performed using Metamorph software (version 7.7.8; Molecular Devices). Cells were bathed in buffer A.
Tirf microscope
Manufactured by Roper Technologies
The TIRF microscope is a specialized imaging tool used for high-resolution observation of biological samples. It employs the principle of total internal reflection fluorescence (TIRF) to selectively excite fluorophores within a narrow region near the sample surface, effectively reducing background fluorescence and improving image contrast.
Automatically generated - may contain errors
2 protocols using tirf microscope
1
Live-cell TIRF Microscopy Protocol
2
Single Particle Tracking PALM of Cav1-mEos2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single particle tracking photoactivated localization microscopy (sptPALM) of MCF and MDCK cells transfected with Cav1-mEos2 was carried out on the Roper Scientific TIRF microscope equipped with an iLas 2 double laser illuminator (Roper technologies), a CFI Apo TRIF 100×
(1.49-NA) objective (Nikon) and an Evolve512 delta EMCCD camera (Photometrics). Images were acquired using Metamorph software (version 7.78; Molecular Devices) at 50 Hz and 16000 frames were acquired per cell. A 405 nm laser was used to photoconvert mEos2, with simultaneous 561 nm exposure to excite the photoconverted mEos2. For stochastic photoconversion of mEos2 molecules, low amount (3-5%) of 405 nm laser and 75-80% of 561 nm laser was used. Data analysis was carried out as previously described 47,48 using PALM-Tracer, a plugin in Metamorph software (Molecular Devices).
(1.49-NA) objective (Nikon) and an Evolve512 delta EMCCD camera (Photometrics). Images were acquired using Metamorph software (version 7.78; Molecular Devices) at 50 Hz and 16000 frames were acquired per cell. A 405 nm laser was used to photoconvert mEos2, with simultaneous 561 nm exposure to excite the photoconverted mEos2. For stochastic photoconversion of mEos2 molecules, low amount (3-5%) of 405 nm laser and 75-80% of 561 nm laser was used. Data analysis was carried out as previously described 47,48 using PALM-Tracer, a plugin in Metamorph software (Molecular Devices).
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!