Expansion microscopy turns the attention to the specimen. It achieves high-resolution images via a chemical rather than optical approach. Preserved specimens are physically enlarged within a swellable hydrogel to allow 3D nano-imaging using conventional microscopes. Tuning the sample may sound tempting, but it comes with some relevant drawbacks. Details >
Knowledge Base
Have you ever wondered how superresolution microscopy works? What’s the difference between STED, STORM, and MINFLUX? What is “resolution” and what is a “PSF”? What is so special about the STEDYCON? Read on to find out.
If you have any suggestions, questions or ideas for our knowledge base, we would be very happy to hear from you.
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How does STED work?
You have heard of STED but don’t have a clear idea how it overcomes the diffraction-limited resolution of confocal microscopes? You maybe even think it to be somewhat complicated? In fact, it isn’t. It’s just physics, smartly applied. Details >
The donut-shaped de-excitation beam is one of the most important practical ingredients for superresolution STED microscopy. But how do you put a hole into a beam of light? Surprisingly, it’s not that difficult if you know how to do it, but it’s very difficult to get it right in practice. Details >
What has to be inside a STED microscope to achieve superresolution? How does its hardware differ from a confocal setup? (Hint: Not very much.) And what does that mean for the user? (Many good things.) Is handling a STED system any more complicated than using a confocal? (Not really.) Important questions – here are some in-depth answers. Details >
How the donut changed the world
For over a century, we stood at the edge of microscope resolution and cursed the inexorable blur of diffracted light. Instruments improved, but the fog never lifted. Then, one man stopped trying to control how light behaves. Armed with a donut-shaped laser beam, he instead commanded where it shines and untethered resolution forever. Details >
How to correct for aberrations in light microscopy
Aberrations can give microscopists a hard time. They belong to microscopy like pathogens belong to life. There are ways to bring diverted rays back on track, but some are better than others. The question is: deformable mirror or correction collar? Details >
Every technique that allows to observe cells is more or less invasive and fluorescence microscopy is no exception. Many imaging situations profit from a reduction in light dose as provided by FLEXPOSURE adaptive illumination. Details >
MATRIX STED is the next level of STED microscopy – combining superior resolution with outstanding signal quality and clarity. Details >
MINFLUX reaches unprecedented spatio-temporal resolution in light microscopy and provides 2D and 3D localization precisions in the single-digit nanometer range. Details >
Why do superresolution microscopists love alpacas?
It is a very simple yet very important fact: the localization precision of any superresolution microscope can only be as good as the size of the fluorescent staining allows. In other words, when your fluorescent dye is too big or too far away from the protein you want to label, you will never be able to reach a resolution that is higher than this offset. The good news is: there are ways to reduce the offset between target protein and fluorescent label. And one of these are nanobodies. Details >
Let the cells shine with immunofluorescence labeling
The most versatile and therefore most common strategy to bring the dye to the sample is immunofluorescence. In case you always wanted to know how immunofluorescence works and which properties of antibodies make it so powerful and at the same time define its limits! Details >
The combination of STED microscopy and PAINT circumvents the physical limitations of current labeling technology. Details >
Photon numbers from the emitting fluorophore. Width of the PSF. How do they impact the resolution of a microscope? Here’s a simple graphic that lays out those effects. Details >
For all the talk about criteria and definitions, measuring the resolution of a microscope is more nuanced than you’d think. The scales at which microscopes operate today are subject to noise and background that obscure and distort signals. What you use for the measurement can make a big difference. The second article in our "Resolution" series. Details >
Are you surprised that the very nature of light caps the resolution that we can achieve in microscope images? Luckily, there are workarounds to this limit. These workarounds push the amount of detail in an image by manipulating precisely where and when fluorophores are allowed to emit. As such, they provide us with a completely new set of tools to shrink the distance between two points while still being able to resolve them. Details >