Understanding research compliance at Janelia when you’re not at Janelia is not always easy. But this blog post will help walk you through what to expect prior to arrival at the AIC and how we will help you prepare.
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.
Needless to say that, just like in any type of microscopy, low signal samples in general do not work well for SIM. Besides that, there are other factors unrelated to the general signal level that will determine if a sample, either fixed or living, is suitable for SIM. The following is by no means a complete list of those factors.
As mentioned in an earlier post, the lattice light-sheet microscope (LLSM) requires special consideration when preparing and mounting samples for imaging. Even after properly mounting a sample for imaging, the raw data produced by the microscope can be hard to intuitively understand due to the angle between the detection objective lens and the sample stage.
One of the most common questions we receive at the AIC is: what exactly constitutes a strong proposal – what does the committee look for? In this blog post, we will explore what makes a proposal stand out favorably amongst its competitors in each review process, and what most of the successful investigators have done to get it right.
The lattice light-sheet microscope (LLSM) is a powerful optical tool for observing rapidly-occurring processes in live samples. This technique offers multiple advantages over more common optical microscopy methods, like spinning-disk confocal, with improved optical sectioning, reduced photobleaching, and minimized phototoxicity. This blog post will cover the most common model systems and the steps we take to mount and adapt the sample for lattice light-sheet microscopy.
The inherent tendency of fluorescent molecules to photobleach has been turned into a very powerful tool for the study of molecular trafficking. By targeting a region of interest for laser-mediated photobleaching, biologists can study how fast the molecules go in and out of the bleached zone and deduce molecular turnover rate from the fluorescence recovery. If a certain type of molecules turns over very fast within the bleached zone, the fluorescence recovery will be rapid.
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).