3D Photoactivated localisation microscopy (PALM) & Stochastic Optical Reconstruction Microscopy (STORM).
Fluorescence single molecule localisation microscopy of fluorescently tagged protein (PALM) or conjugated antibody (STORM) for 20nm precision in fixed samples.
STORM employs the photoswitchable, aka ‘blinking’, properties exhibited by some fluorophores. Allowing a different subset to be ‘switched on’ at any one time allows for temporal separation of the fluorescent probes that would otherwise spatially overlap and render localisation of their respective centres difficult. Many imaging cycles are conducted, in which fluorophores are activated, imaged and deactivated, with each cycle capturing a different sub-population of the fluorescent probes. The centres of all of these fluorescent ‘blinks’ can be determined and then reconstructed to form a super resolution image.
- 3D super resolution imaging of fixed samples
- Cells expressing fluorescently labelled proteins (PALM)
- Cells stained with labelled antibodies (STORM)
- Water 60x objective, NA 1.20 (UPlanSApo)
- 405 nm
- 488 nm
- 532 nm
- 561 nm
- 647 nm
L.C.Miall 5.07, Faculty of Biological Sciences