Super Resolution RCM1 / RCM2  Promo: Learn More!

How super-resolved fluorescence microscopy led to a Nobel Prize

In 2014, Eric Betzig, Stefan Hell and William Moerner got the Nobel Prize in Chemistry, for their work in developing super-resolution fluorescence microscopy. The three scientists developed different techniques for achieving super-resolution imaging. This includes stimulated emission depletion (STED) microscopy (developed by Hell), photo-activated localization microscopy (PALM, developed by Betzig) and single-molecule spectroscopy (developed by Moerner). But what does this mean and where does their theory stand in 2023? Let’s find out.

The beginning of super-resolution fluorescence microscopy

Until the end of the nineteenth century, fluorescence microscopy was, as Ernst Abbe (Abbe, 1873) and Lord Rayleigh (Rayleigh, 1896) defined it: “Diffraction limited”. It was impossible to distinguish between two objects that were laterally closer than about half of the used wavelength and even further apart in depth (around 500nm). Taking into account that the commonly used wavelengths are on the visible spectrum, lateral resolution could only resolve structures above 200nm (far blue) and 350nm (far red).

In the 1990s and early 2000s, new approaches to surpass Abbe’s limit started appearing. A timeline of these inventions can be found right here. In those first years, Eric Betzig contributed with his research in near-field scanning optical microscopy (NSOM), which can be found in this study. But the NSOM approach had limited applicability in the field of biology.

Early far-field advances started appearing in this period as well. Lateral resolution improvements started with Confocal microscopy, improving sectioning through light rejection. It was followed by the theoretical foundations of stimulated emission depletion (STED), single-molecule localization microscopy (SMLM) and structured illumination microscopy (SIM). The inferior axial (z) resolution in epifluorescence microscopy was initially addressed by using opposing objectives in confocal (4Pi) and widefield microscopy (I5 M).

The impact of super-resolution fluorescence microscopy techniques in biology

At the same time, the theoretical foundations of super-resolution microscopy (SRM) got a push in advances on fluorophores, lasers, computers and detectors technology. This helped the field to move forward. New techniques were perfected and became the pillars of super-resolution. These include: STED, SIM, PALM, and dSTORM.

These techniques continued to be developed and improved upon, with new methods such as DNA-PAINT and super-resolution fluorescence lifetime imaging microscopy (FLIM-SR) emerging in recent years. Additionally, the use of machine learning and image processing techniques, such as deep learning, may be used to improve the accuracy and efficiency of super-resolution microscopy. These techniques have revolutionized the field of microscopy and have led to many new discoveries in areas like cell biology and neuroscience.

The future of Super-resolution microscopy

There are several potential ways to improve super-resolution microscopy in the future. Long term super-resolution live cell imaging can be greatly improved upon. One of the major drawbacks used to be the intensity of the laser power, which needed to excite the sample to be high enough to cause phototoxicity. This used to hamper long term live imaging, but that’s luckily no longer the case.

The speed of SRM can also be a venue to explore. Nowadays, technology commonly needs long processing times and sample preparation. Finally, the integration of super-resolution microscopy with other imaging modalities, such as electron microscopy, may allow for even more detailed and accurate imaging of biological samples.

What Confocal.nl technologies can offer

Confocal.nl solutions focus on providing the highest resolution possible in a setting that allows long term live cell imaging. To do this, we use a combination of low laser power (nanoWatts at the sample plane) together with an increase in the signal to noise ratio (SNR) of four times higher compared to conventional confocal systems.

Our RCM technology can convert your existing widefield fluorescence microscope into a super resolution confocal system, with high sensitivity.

To get more information about our RCM products, you can download our brochure to explore how we improve your imaging experience.

Confocal demo form image

See for yourself how we improve your experience

Request your
personalized demo

  • Hidden

This website uses cookies

With these cookies, we and third parties can collect information about you and your online behavior, both within and outside our website. Based on this, we and third parties tailor the website, our communications, and advertisements to your interests and profile. You can find more information in our cookie statement.

Accept Reject Preferences

This website uses cookies

With these cookies, we and third parties can collect information about you and your online behavior, both within and outside our website. Based on this, we and third parties tailor the website, our communications, and advertisements to your interests and profile. You can find more information in our cookie statement.

Functional cookies
Arrow down

Functional cookies are essential for the proper functioning of our website. They allow us to enable basic functions such as page navigation and access to secure areas. These cookies do not collect personal information and cannot be disabled.

Analytical cookies
Arrow down

Analytical cookies help us gain insight into how visitors use our website. We collect anonymized data about page interactions and navigation, enabling us to continuously improve our site.

Marketing cookies
Arrow down

Marketing cookies are used to track visitors when they visit different websites. The goal is to display relevant advertisements to the individual user. By allowing these cookies, you help us show you relevant content and offers.

Accept all Save
Confocal demo form image

See for yourself how we improve your experience

Request your
personalized demo

  • Hidden

Contact us

See for yourself how we improve your experience

Request your personalized demo

In about 45 minutes to an hour, our experts show you how our products perform live.

Become a distributor

Contact a distributor

Contact us