Common Applications
STORM: |
Stochastic Optical Reconstruction Microscopy is a super-resolution microscopy technique utilising a TIRF architecture to illuminate the first 100-200 nanometers of a sample and image single fluorescent molecules as they are first converted from a dark to active state by an appropriate buffer solution, before bleaching a short time later. At any time only a very small portion of the fluorophores are active and with each dark-active-bleach cycle more and more of the underlying sample is revealed at up to 10x the resolution of conventional techniques. |
PALM: | Photo-activated localization microscopy is a super-resolution microscopy technique where the principle of detection and analysis is the same as STORM, but in this case it is not anti-body based labelling and does not require special buffers, but rather requires use of photo-activable genetically encoded proteins. |
SIM: |
This method involves illuminating a fluorescent sample with an illumination pattern, here a series of light and dark bands. After acquiring a series of images with the illumination pattern in different orientations, spatial information about the sample that normally would be lost, can be yield. It requires at least 3 rotations (optional 5) and 5 phase images for a total of 15 (or 25) images in order to reconstruct one super-resolution image. As such it makes this technique compatible with live cells imaging but for intermediate to slow dynamical processes. |