Dual-color SRRF imaging was performed on a spinning disc confocal microscope (14 (link)). HMRef and Alexa Fluor 647 phalloidin excitation was conducted with a 488-nm/150-mW diode laser (LM-488-150, Andor) and a 637-nm/140-mW diode laser (LM-637-140, Andor), respectively. The two lasers were fiber coupled (seven-line laser combiner, multimode ×2, single mode ×1; LC-ILE-700-M2-S1, Andor) to a spinning disk confocal unit (CR-DFLY505, Andor) equipped with a multiband dichroic mirror (DFly laser dichroic for 405/488/561/640). The fluorescence was processed with appropriate filter sets for HMRef (TR-DFLY-F525-050, Andor) and Alexa Fluor 647 (TR-DFLY-F700-075, Andor) to capture fluorescence images with a charge-coupled device (CCD) camera (iXion Life 888, Andor), driven by Fusion software (version 2.0 for Fig. 2 and version 2.2 for fig. S19; Andor). Images were taken using a 60× objective (APON60XOTIRF, numerical aperture (NA) 1.49, Olympus], mounted on an inverted microscope (IX83, Olympus), and equipped with Z-drift compensator (IX3-ZDC2, Olympus).
Protocol detail
Multi-color Super-Resolution Imaging Protocol
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : Japan Agency for Medical Research and Development
Other organizations : Kyoto University, Japan Science and Technology Agency, Keio University, Yamaguchi University
Top 5 similar protocols
Variable analysis
independent variables
- Excitation with a 488-nm/150-mW diode laser (LM-488-150, Andor)
- Excitation with a 637-nm/140-mW diode laser (LM-637-140, Andor)
- Fluorescence images captured with a charge-coupled device (CCD) camera (iXion Life 888, Andor)
- Spinning disc confocal microscope (
14 (link)) - Fiber coupled (seven-line laser combiner, multimode ×2, single mode ×1; LC-ILE-700-M2-S1, Andor) to a spinning disk confocal unit (CR-DFLY505, Andor) equipped with a multiband dichroic mirror (DFly laser dichroic for 405/488/561/640)
- Fluorescence processed with appropriate filter sets for HMRef (TR-DFLY-F525-050, Andor) and Alexa Fluor 647 (TR-DFLY-F700-075, Andor)
- Images taken using a 60× objective (APON60XOTIRF, numerical aperture (NA) 1.49, Olympus], mounted on an inverted microscope (IX83, Olympus), and equipped with Z-drift compensator (IX3-ZDC2, Olympus)
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!