Hi,
I was wondering if it is known if STORM microscopy could be used to obtain super-resolution images of sfGFP or mVenus tagged proteins in cells.
I know it is more common to use fluorescent probes.
Any thoughts on this?
Thanks,
Florian
Dear Florian Pruckner ,
STORM (Stochastic Optical Reconstruction Microscopy is a super-resolution microscopy technique that can provide images with a resolution beyond the diffraction limit of light. It relies on the stochastic blinking behavior of fluorescent probes to localize individual molecules with high precision, enabling the reconstruction of super-resolved images.
In principle, STORM microscopy can be used to obtain super-resolution images of any fluorescently labeled protein in cells, including those tagged with sfGFP or mVenus. However, the quality and suitability of the images may depend on several factors, such as the expression level and distribution of the tagged protein, the labeling density and photostability of the fluorescent protein, and the efficiency of the labeling and imaging protocols.
While fluorescent probes are often used in STORM microscopy because of their high photostability and brightness, fluorescent proteins such as sfGFP and mVenus can also be used as labels. However, these proteins may exhibit more complex photophysical behaviors than synthetic probes, which could affect the quality of the images. Additionally, it may be necessary to optimize the labeling and imaging conditions to obtain the best results.
Overall, while the use of sfGFP and mVenus as labels in STORM microscopy is less common than synthetic probes, it is feasible and could be a valuable tool for investigating the distribution and dynamics of these proteins in cells.
No, these won't work. You have to be able to manipulate the on/off states of the dye either through photoactivation or stochastic photoswitching in the appropriate buffer. These labels do not have the appropriate photophysics to "blink" on and off as required for STORM. Other methods such as SIM or SRRF could work with these labels.
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