3D Super Resolution Imaging (STORM/PALM)
15 Oct 2012





​Zeiss Elyra PS1: STORM/PALM in the same microscope as structured illumination imaging, with an EMCCD detector. 3D imaging possible using the double-helical point-spread function method. Activation/excitation wavelengths are 405, 488, 561 and 640 nm.

For more details please contact Dr. Lin Wang.
​​Technique and applications

Single-molecule localisation super-resolution imaging. Down to 20 nm resolution in xy and 50 nm in z over 1.5 µm z range. z-resolution using phase ramp or biplane methods.
TIRF, HILO and epi-illumination.​

Biological and medical research, material science, chemistry

Lasers: Multicolour imaging with 488, 561, 642, 750 nm excitation and 405 nm activation

Detectors: EMCCD and sCMOS cameras

Objectives: Elyra- 63x oil and water immersion,100x oil and silicone immersion

Single molecule localisation microscopy is a family of super-resolution microscopy techniques, including stochastic optical reconstruction optical microscopy (STORM) and photoactivated localisation microscopy (PALM). A series of images (typically 5-15,000) are acquired, each of which contains a number of diffraction-limited spots. Each spot is assumed to be symmetrical and Gaussian in profile, allowing its centre to be localised down to ~10 nm. The resulting co-ordinates are aggregated and can be rendered as a super-resolved image. The challenge is to switch the fluorescent molecules on and off such that each appears in as few frames as possible, usually using a combination of a 405 nm activation laser and an appropriate buffer solution.

InstrumentZeiss Elyra PS1
Laser wavelengths (nm)405 (for activation only), 488, 561, 642
Excitation power densityModerate, depends on field of view
Compatible objective lensesx100 oil immersion, x100 silicone oil immersion, x63 oil immersion, x63 water immersion
Compatible dyesRestricted, depends on blinking characteristics and photon flux
Compatible sample mounts#1.5 cover slip on microscope slide or glass bottom Petri dish
ConfigurationInverted microscope
Axial resolution methodPhase ramp
Multi-wavelength imagingUp to three in 2D or 3D, always sequential
Temperature control21-37°C
Lateral resolution
~20 nm (highly dependent on signal to noise/background ratios and labelling density)
Axial resolution~50 nm (highly dependent on signal to noise/background ratios and labelling density)
Axial range1-1.5 microns
Temporal resolution
4 minutes (max)

Microscope can be used with epi or TIRF illumination. 

Contact: Wang, Lin (STFC,RAL,CLF)