Breaking through traditional optical diffraction limits: the new generation of Nanoimager makes it easy to achieve super-resolution fluorescence imaging

In recent years, with the development of single-molecule fluorescence imaging technology of living cell system, the study of membrane protein single molecule, especially receptor kinetics, has become one of the most active research directions in the field of single molecule research. The super-resolution imaging technology developed in recent years has higher resolution and higher positioning accuracy than traditional optical microscopes because it can break through the optical diffraction limit.

The Nanoimager, the latest super-resolution fluorescence microscope from Oxford Nanoimaging, developed by the team of professors of the University of Oxford, Achillefs Kapanidis, is the world's first large-field single-molecule FRET microscope with super-resolution at a single molecule. Research fields such as tracing, live cell imaging, protein interaction, and 3D imaging play an important role.

Nanoimager main technical features

♦ Horizontal resolution <20nm; vertical resolution <50nm
♦ Stability: <1 μm/K drift; <1 nm (1 Hz to 500 Hz) amplitude ♦ Supports simultaneous two-color imaging and sequential four-color imaging ♦ Class 1 laser, safe to use
Figure 1 Nanoimager super-resolution imaging
The Nanoimager uses PALM/dSTORM technology and photoactivated localization microscopy (PALM) to obtain the center position of the fluorescent molecule with ultra-high precision (nanoscale) using a single-molecule localization algorithm combined with the shape of the optical system Airy disk, and then using CCD The signal is collected and transformed to finally obtain a super-resolution image with a resolution of 20 nm.
Nanoimager main application case

1, single molecule FRET
FRET is a non-radiative energy transfer between two fluorescent molecules, reflecting the molecular spacing of the two (generally occurring at a 2 - 10 nm spacing). Nanoimager is the world's first commercial instrument for large-field single-molecule fluorescence resonance energy transfer (smFRET). Its key functions for smFRET include: simultaneous two-color imaging; single-molecule scattered light intensity and overall average real-time analysis; High-throughput imaging of thousands of single molecules and the function of alternating fluorescence excitation (ALEX) smFRET to quantify stoichiometry and FRET efficiency. Figure 2 is a smFRET used to study the kinetics of a single DNA Holliday crossover.
Figure 2 Detecting the real-time conformational changes of Holliday crossings (HJs) using smFRET
2, single molecule tracer
The Nanoimager can simultaneously trace single molecules in a cell or purified sample in both channels (Figure 3) and calculate the diffusion coefficient. The diffusion coefficient of molecules in cells can be traced, such as enzymes or proteins can be traced by the reaction of drugs and antibiotics. A low diffusivity can mean an interaction or combination of a labeled molecule with another molecule or structure.
The Nanoimager directly reflects the diffusivity and estimated size of the fluorescent particles in the purified sample, with sensitivity (single fluorescent molecular level) and specificity (two-color labeling can significantly reduce the possibility of detecting impurities).
Figure 3 Nanoimager two-color tracking single molecule / particle
3, larger field of view imaging

Each imaging channel of the Nanoimager has a large field of view of 50 μm x 80 μm and is evenly illuminated for high-throughput imaging of single molecules or cells and rapid data collection. Figure 4 shows imaging of different phenotypes of mutant E. coli cells at a rate 10 times faster than other techniques. In order to obtain reliable results for different phenotypes, a large number of cells need to be compared. Using a Nanoimager with a large field of view that autofocuses and automatically acquires data can dramatically speed up the entire experiment with speed and throughput. Combining large field of view with super-resolution imaging is a unique advantage of the Nanoimager.
Figure 4 Nanoimager's large field of view enables high-throughput imaging at high resolution
Super-resolution fluorescence microscopy has become an important tool for biomedical research with its unique advantages. If you want to know more about the technology and application of Nanoimager , please call 010-85120280 for consultation, we will give you a satisfactory answer as soon as possible!
Related products and links
1. A new generation of super-resolution fluorescence microscope (NEW): http://?id=443
2. LaVision BioTec Light Film Illumination Microscope: http://?id=434
3. Two-photon fluorescence microscope: http://?id=47
4, LVEM5 desktop transmission electron microscope: http://?id=244

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