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 >
Since the 1990s, confocal microscopes have been a staple in labs visualizing biological or material specimens. The development of STED microscopy prompted the question: how does the established confocal microscope compare to the (now not so) “new kid on the block”? Details >
Structured illumination microscopy offers some advantages over confocal, most notably increased resolution. Comparing it to STED, however, reveals its limitations. Details >
Which microscope has the best resolution?
The elctron microscope achieves the highest magnification and resolution. But does "highest" always equal "best"? Well, that depends on what you want to do with the resolution. Details >
Confocal microscopy offers superior optical sectioning. But what is that exactly? And what about other ways to get rid of the background, such as array-based detectors like the MATRIX? Details >
For centuries, conventional light microscopy was and continues to be the workhorse of labs to visualize cells and cellular details. But the advent of electron microscopy brought about a new level of detail. Let's take a closer look at the two techniques. Details >
Deep and clear: where confocal beats out wide-field microscopy
Confocal microscopes were designed to get rid of background signal. How do they work? And when do you know it’s time to use one? The answer is in the pinhole. Details >
Fluorescent labeling strategies have become more and more sophisticated and offer ever-new options to improve microscopic imaging. Among the latest are exchangeable HaloTag ligands that put an end to photobleaching for good. 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 >
Superresolution for biology: when size, time, and context matter
The spatial resolution achievable with today’s light microscopes has unveiled life at the scale of individual molecules. Size is no longer a barrier to seeing biology at the most fundamental level. But life is not static. It emerges from movement and change. How do superresolution technologies hold up to the challenges of documenting dynamic biological mechanisms? 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 >