STEDYCON:

A sleek, black-and-orange box transforms your widefield microscope into a confocal and a superresolution STED instrument and your exploration of subcellular structures into a seamless, discovery-rich experience. Carefully designed with masterly engineering, STEDYCON breaks the stereotype of the finicky, hard-to-use scope. It opens new possibilities at the press of a button for any user and almost any location. How does it do it? The secret’s in the box.

ease-of-use in a shoebox

Human ingenuity

Human ingenuity commands admiration. We coax living entities – from elephants to viruses – into doing our bidding. We source out physical and mental work to machines. We sling people across the globe in thin-walled, stratospheric aircrafts. We receive pictures of the universe from a telescope 1.5 million kilometers from the Earth. Although it is the blink of an eye in the grand scheme of things, human history is a maelstrom of innovation.

All kinds of human motives drive that ingenuity. Need. Pleasure. Curiosity. Sharing. Greed. Fueling the adoption of its products, however, is one quality: ease-of-use. The press of a button that unleashes transactions, knowledge, and emotions is the name of the game.

Engineering ease-of-use into cutting-edge microscopy

The forces that drive technology adoption are no different in the science lab. Highly specialized instrumentation is limited to core facilities or niche research groups. Specific techniques may require an occasional unorthodox device in some labs, but a staple set of equipment and methods appears on nearly every benchtop, and its acquisition is driven by ease-of-use. Ease of installation. Ease of proficient use. Ease of results. By consequence, if you want to transform new technology into an everyday tool, you need to design it for one thing: ease-of-use.

That thought sparked the engineering feat that brought superresolution microscopy into the average science lab. With stimulated emission depletion (STED) microscopy and other techniques already enabling sub-diffraction resolution, it seemed unreasonable that researchers curtail their ability to visualize sub-cellular structures by using diffraction-limited microscopes. Clearly, adding STED to the standard constellation of lab equipment would require more than outstanding resolution. The problem, developers reasoned, was that Nobel-prize technologies are like Formula 1 race cars that only high-performance drivers can maneuver. So, their strategy was to distill that Nobel-winning “race car” into a shoebox that everyone could “drive.” STED would become a powerful upgrade to any widefield microscope with plug-and-play operation, intuitive handling, and rapid outcomes. The product was STEDYCON.

STEDYCON: a confocal and STED microscope

The STEDYCON upgrades your existing widefield system to a confocal and a superresolution STED microscope with a resolution down to 30 nm.

So, what does it look like?

At first glance, STEDYCON is a black-and-orange casing about the size of a shoebox that sits on the camera port of a microscope. It is compact, unobtrusive, and sturdy. The plain exterior presages its ease-of-use but belies what lies within. Carefully planned engineering and design transform your microscope into a multicolor confocal and 2D STED system. It’s like plugging your scope into an amplifier and being rewarded immediately with power.

To begin with, the engineers tackled the big nuisances that make superresolution microscopy finicky. STEDYCON requires no alignment of the excitation and depletion lasers. They are aligned by design. With a novel optical arrangement that sends all laser beams through the same fiber, the system is more stable than other STED microscopes where beams travel separately. That inherent stability shortens the time to initiate imaging and simplifies installation, which takes only minutes. Furthermore, STEDYCON includes a scanner technology that allows it to operate with any widefield microscope, equalizing performance across platforms and precluding the need for dedicated equipment.

The engineers also revamped the autofocus. Unlike conventional optical instruments that use a dichroic mirror to couple the autofocus laser with the imaging beam path, STEDYCON runs the two beams parallel but segregated. Thus, autofocus in STEDYCON never interferes with imaging, which can produce fluorescence loss or distortions. With the simple press of a button, STEDYCON guarantees drift-free focus for extended imaging sessions up to several days. Also, as STEDYCON is equipped to control motorized stages, it automates the recording of multiple positions in a sample or a tile scan. The user is thus released from sitting at the microscope and can easily capture the whole sample or navigate across it to find objects of interest.

Secondly, STEDYCON makes superresolution microscopy a research technique for everyone. With minimal training, even novices to microscopy can produce high-quality STED images with a few intuitive manipulations of a browser-based control system with a user-friendly interface. Whichever the chosen procedure, users commonly visualize structures at a resolution of 30 nm.

Finally, STEDYCON makes no compromises on sensitivity or dynamic range of detection. The avalanche photo detectors (APDs) in STEDYCON have superior quantum efficiency. Thus, they reliably collect photons even under low-signal conditions, like when labeling densities are minimized to preserve physiological conditions in samples. This feature ensures clear images and exceptional signal-to-noise ratio, even under high-signal conditions.

The sway of ease-of-use

The greatest testament to the performance and value of STEDYCON is its growing user base. Being frame-agnostic, STEDYCON complements microscopes of various makes and models. Exceptionally stable, it breaks with convention by operating flawlessly in dedicated labs, trade fair floors, living rooms, and even campsites. And STEDYCON users are a new generation of innovators leveraging a level of microscopy previously reserved for “Formula 1 drivers”. Swayed by easy access to unprecedented detail, they add diversity to the where, when, and how superresolution is used and uncover a myriad of insights hidden behind the low resolution of standard microscopes. There, at the frontiers of science, those discoveries unlock new lines of research as the boundaries to how we study structure, movement, and interactions at the smallest scale are lifted. STEDYCON’s ease-of-use in a shoebox is a portal to breakthroughs in an infinite world.

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How does STED work?

A donut-shaped STED beam confines the fluorescence to a sub-diffraction sized area

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

Nobel laureate Stefan W. Hell shows a donut, the symbol for his groundbreaking idea of a donut-shaped laser beam.

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 >

PALM and STORM are often used as synonyms, and in fact they have a lot in common. But there are slight differences that can be important for your application. And then there are other superresolution techniques, too – like STED and MINFLUX. 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

How to correct for aberrations in light microscopy. Deformable mirror vs correction collar!

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 >

Why do superresolution microscopists love alpacas?

Immunofluorescence staining with alpaca nanobodies

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 >

Superresolution for biology: when size, time, and context matter

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 >

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 >

Dr. Joachim Fischer. Caption: Physicist, knows how to fit a STED and a confocal microscope in a shoe box

How did you manage to fit both, a STED and a confocal microscope, in a shoe box?

When designing STEDYCON, abberior aimed for a super compact, super stable and super easy to use STED microscope, without compromising on performance.

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STEDYCON upgrades existing widefield systems to confocal and superresolution STED microscopes

STEDYCON

Just upgrade your existing widefield system to a confocal and STED microscope with a resolution down to 30nm. Enjoy fine imaging at the push of a button!

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Dr. Janina Hanne. Caption: Biologist, maybe the only person with a STEDYCON in her living room

Can everyone have a STEDYCON in their living room?

Yes, of course! Our STEDYCON is robust, reliable and easy to use. In the smallest of spaces you can run confocal and superresolution STED experiments.

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abberior LIVE

abberior LIVE dyes offer exceptional labeling options. All the advantages of organic dyes, but in living cells. Unraveling biological functions is now a breeze!

Details >