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The closest focusing distance i.e. 9cm/ 3.54” makes create stronger background bokeh effect. It is also useful when getting a close up on your subjects.

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 >

so alpha = 2*arctan(d/2f). Now you can simply calculate it for every lens and format. All of that of course when focused to infinity. For ...

When it comes to superiority in resolution, the simplest question to answer is which type of microscope achieves the highest magnification and resolution. Hands down, that is the electron microscope. In fact, the Guinness World Record for the highest resolution is held by an innovative, algorithm-driven version of the electron microscope that visualized single atoms of oxygen, scandium, and praseodymium. But does “highest” always equal “best”?

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Laowa 6mm T2.1 Zero-D MFT Cine is a tiny and light lens suitable for everyday filming. The lens weighs around 180g/ 6.35oz and is measured to be ~Φ61 x 53mm/ 2.40 x 2.08”.

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 >

The constant maximum t-stop T2.1 is also favorable and flexible to shoot under low-light conditions. It can render blurred background and creamy bokeh as well.

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 >

Biological specimens require a lot of sample preparation for electron microscopy. Why? Because you’re doing the equivalent of sending them into space and then blasting them with high energy. First, the specimen must be fixed. Otherwise, the energy of the electron beam will destroy it. The specimen must also be dehydrated to survive the intense vacuum inside the microscope. Then, many biological specimens are not conductive. As a result, electrons can’t pass through the sample, and you don’t get an image. Making biological specimens conductive involves coating them with a thin layer of metal.

Electron microscopes shoot a concentrated beam of electrons at a target object. An image is produced as the electrons pass through the specimen and are detected. Because the electron wavelength is several thousand times shorter than that of light, the resolving power of an electron microscope is a thousand times greater than that of a light microscope.

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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 >

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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 >

Circularly polarized electromagnetic waves are composed of photons with only one type of spin, either right- or left-hand. Linearly polarized waves consist of ...

However, it's impossible to find a lens without chromatic aberration, since it's a physiological problem of all lenses when exposed to light. Dispersion also ...

Ideal imaging conditions are often compromised by imperfections in the optical path. These can severely compromise a microscope’s performance, unless they are eliminated by RAYSHAPE's deformable mirror. 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 >

In the realm of light microscopy, MINFLUX has repeatedly demonstrated single-digit nanometer resolution and below. The characterization of nuclear pore architecture and mitochondrial protein patterns are just two examples of the technology’s spatial resolving power. This capacity grants a completely new perspective on molecular structure in biological context, revealing the architecture of biomolecules and their interactions.

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 >

With a 121.9° angle of view, Laowa 6mm T2.1 Zero-D MFT Cine becomes the widest rectilinear lens in the Micro Four Thirds market. Filmmakers are able to capture more details in the shot and make it unrestricted in a tight space. It is suitable to be used on drone, FPV drone, vlogging, extreme wide shots etc.. Zcam E2-M4 , BMPCC 4K , GH5 and other MFT camera would fit this lens perfectly.

Structured illumination microscopy offers some advantages over confocal, most notably increased resolution. Comparing it to STED, however, reveals its limitations. Details >

Research that does not require characterizing individual molecules but rather their spatial relation to others has a broader range of microscopy technologies at its disposal. Another step down on the resolution scales makes widefield, confocal, STED, and PALM/STORM microscopy all options (see figure). As super-resolution technologies, STED and PALM/STORM outperform the spatial resolution of diffraction-limited confocal and widefield microscopy by a factor of 10. Commonly discriminating objects at 20 nm, STED is also fast, which has enabled, for example, visualizing the fission and fusion of mitochondria with exceptional clarity.

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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 >

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For STED microscopy, similar sample preparation techniques may be utilized as for conventional microscopy. However, the increase in special resolution requires additional precautions to ensure the structural preservation of the specimen. Details >

MINFLUX reaches unprecedented spatio-temporal resolution in light microscopy and provides 2D and 3D localization precisions in the single-digit nanometer range. 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 >

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 >

A little insight into the advances in virus research made possible by STED microscopy and a hint to were the journey might go. Details >

Rectilinearlens

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 >

As you consider the advantages of resolution for your research, you should have a clear idea of what resolution means. It is distinct from magnification. In light microscopy, it is limited by an immutable property of light. And it is achieved and measured in different ways. Here are two articles to get you up to speed: “What is resolution?” and “How to measure resolution?“

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 >

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Every scientific instrument comes with tradeoffs. Augmented power in one dimension, invariably curtails another. In the case of microscopes, boosting resolution complicates sample preparation and narrows the application spectrum. The best resolution is not always the highest resolution. When you’re looking for the best resolution, consider what you want to see. Most of the time, not only size but also time and context matter.

The combination of STED microscopy and PAINT circumvents the physical limitations of current labeling technology. Details >

Jun 7, 2021 — A technical but important element of photography is depth of focus, which is easily confused with depth of field.

But is it the best resolution? Highest, yes. Best? Well, that depends on what you want to do with the resolution. In biological research, time and context often matter at least as much as size, which is why superresolved light microscopy techniques like STED and MINFLUX can play to their full strength here.

"What are you?" You know what I mean – your eyes look Chinese, or something. No offense. A Newfoundlander allergic to FISH?! I am. Inuk. Irish. Freckled.

Confocal and multi-photon microscopy are used for deep tissue imaging, but misconceptions about their utility have led to their misuse. We’ll plunge into tissue depths to reveal a gap in obtaining sharp images that RAYSHAPE – a solution for dynamic aberration correction – fills with clarity and brightness. Details >

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. Details >

Laowa 6mm T2.1 Zero-D MFT is the widest rectilinear lens you can find in the Micro Four Thirds market. With a 121.9° angle of view and maximum T2.1 t-stop, filmmakers can obtain a unique-to-your-eye perspective even under low-light environment. This lens is also designed as “Zero-D” to deliver excellent image quality. Moreover, its tiny size and 58mm filter thread design make it more convenience and handy to use. It can be used for establishing shots, FPV drone, vlogging and more. MFT cameras like BMPCC4K, GH6, Zcam E2 M4 etc. would fit perfectly.

In fact, the most distinctive feature of MINFLUX is its temporal resolution, which gives it the most advanced tracking capabilities of currently established microscopy technologies – by a long shot. That means that you can watch changes and movement happening inside living cells. The ability to differentiate events that are just a hundred microseconds apart expands the applications of MINFLUX from structural biology and slow processes like gene expression, to diffusion phenomena and even conformational changes of biomolecules. An example of this unprecedented power was the recent tracking of a kinesin-1 molecule walking along microtubules, including the corresponding configurational changes occurring at each step.The movement of kinesin-1 has never been tracked in a living cell before.

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 >

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 >

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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 >

Chris Dadge, David Laing, and Scott Munro (along with Kevin Lee) formed the Bent Spoon Ensemble in 2002, and after a few years ...

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 >

Today’s high-end fluorescence microscopy is unthinkable without lasers. Reason enough to take a closer look at these sophisticated light sources. Details >

Clearly, the hostile environment of an electron microscope precludes working with live or unfixed samples. So, if you’re interested in the movement, changes, and context that constitute life, a slight downgrade in resolution is likely your best bet. Enter the world of super-resolution microscopy.

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Perhaps the most important differentiators of STED are its economic photon budget and easier sample preparation and data analysis compared to PALM/STORM. In fact, as a mature technology, STED microscopes are as easy to use as standard confocal microscopes. Furthermore, STEDYCON uniquely merges all three parameters – strong resolution, intuitive usability, and broad flexibility – into one exceptional instrument: a sleek box with a favorable price tag that transforms your existing widefield microscope into a confocal and a full-fledged STED instrument.

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 >

This lens is named as “Zero-D”. The newly devised optical design contains 13 elements in 3 groups with two aspherical glasses, delivering outstanding image quality with minimal distortion. The close-to-zero distortion (Zero-D) design is extremely useful when shooting architectural, aerial and indoor footage.

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 >

Today’s research microscopes are increasingly powerful, modular, and combinatorial. There’s a lot of options out there. While the price is unquestionably a deal-breaker for purchase, a more helpful criterion is value. Details >

MATRIX STED is the next level of STED microscopy – combining superior resolution with outstanding signal quality and clarity. Details >