In a previous blog post, we discussed probe selection for single molecule localization microscopy (SMLM) techniques such as PALM and STORM. This blog post continues earlier discussions to give some practical guidance toward both probe selection and imaging considerations when performing SMLM across multiple channels.
The last 10 years has seen an explosion of techniques aimed at creating biological fluorescence images with unsurpassed clarity via super-resolution microscopy. Among these various techniques that circumvent the diffraction barrier, there is a group of methods that can be termed single-molecule localization microscopy (SMLM). This blog introduces the conceptual basis, and gives pointers for selecting these fluorescent tags for use in SMLM.
For many researchers, the prospects afforded by super-resolution (SR) imaging are both vast and exciting. Unfortunately, the dizzying array of acronyms reported in the literature, including PALM, STED, SIM, GSDIM, RESOLOFT, iPALM, dSTORM, among many others, can leave researchers unsure as to the optimal technique for their specific application(s). This article will provide a brief overview of the major classes of SR techniques, with suggested reading for further details. The second part of the article will outline a practical comparison of the two SR systems currently available at the Advanced Imaging Center (AIC).