A custom-built PALM/STORM super-resolution microscopy platform under S2 containment enables imaging of fluorescent proteins mEos (PALM) and organic fluorophores Alexa 532 and Alexa 647 (STORM) at a spatial resolution of 20-30 nm (in 2D).
The equipment consists of a set of laser modules that are custom-coupled into a Zeiss Observer Z1 microscope. The microscope is equipped with 100X NA 1.46 oil immersion objective (planachromat). It contains (i) a fully-automated control of filters and objectives, (ii) a motorized x-y stage together with a z-piezo, (iii) active focus stabilization and (iv) both TIRF and widefield illumination.
The PALM/STORM setup is available to other institutes within the campus and can be used by S2-biosafty level authorized people.
Training to allow researchers to work independently with the equipment will be provided by Walter Muranyi from the Department of Infectious diseases, Virology. Due to the complexity of the equipment, extensive trainings are necessary.
However, we recommend using the equipment in the framework of a collaboration, which will optimize research results. Additionally, protocols will be provided from the institute. User of the equipment will not be charged.
Merge of a PALM image for Gag.mEosFP (green) and the corresponding dSTORM image of Env stained with Alexa Fluor 647 (red). Scale bar, 1 µm.
Individual assembly sites from the boxed regions indicated in (A). Merged super-resolution images (left panels), the dSTORM Env Alexa Fluor 647 image (middle panels) and individual Alexa Fluor 647 localizations from all images recorded in the defined area as black dots. The black circle represents the Gag cluster (right panels). Scale bar, 100 nm.