In the ever-evolving world of microscopy, the Vutara VXL by Bruker is making waves with its innovative approach to single-molecule imaging. This cutting-edge technology, unveiled recently, promises to transform how researchers conduct high-throughput 3D super-resolution imaging, particularly in molecular biology settings.
The Vutara VXL stands out as a versatile platform, enabling precise localization of DNA, RNA, and proteins. Its unique combination of bi-plane detection and top-hat illumination, paired with a silicon-immersion objective, allows for streamlined DNA-PAINT multiplexing through the PlexFlo platform. This integration ensures that researchers can achieve robust imaging results with unparalleled ease and precision.
Advancements in Single-Molecule Localization Microscopy
Single-molecule localization microscopy (SMLM) has long been a cornerstone of super-resolution techniques, offering detailed insights into cellular structures. Unlike traditional methods bound by diffraction limits, SMLM achieves resolutions as fine as 10 nm by pinpointing individual fluorescent molecules. This leap in resolution is crucial for observing intricate subcellular structures, from mitochondria to viruses.
To put this into perspective, a high-end confocal microscope can image microtubules, approximately 25 nm in diameter. However, due to diffraction limits, these structures often appear blurred, measuring between 200 and 250 nm in final images. SMLM overcomes these limitations, providing clarity and detail previously unattainable.
Design Innovations for Enhanced Stability and Compactness
The Vutara VXL’s design is a testament to Bruker’s commitment to innovation. As a benchtop optical microscope optimized for single-molecule localization, it features a larger field of view essential for omics applications. The system includes five powerful laser lines for illumination, a flat illumination profile, and an integrated multiplexing platform, all housed within a compact design.
Key components such as a laser module, an electronics control e-box, chillers for thermal management, and a fluidics module for multiplexing are tightly integrated. This setup, along with active temperature control and an autofocus module, ensures remarkable system stability, with Z-drift typically staying around 200 nanometers across multiple days of acquisition.
Bi-Plane Detection System: Enabling 3D Localization
The bi-plane detection system is at the heart of the Vutara VXL’s ability to achieve 3D localization. By capturing two distinct planes on a sensor simultaneously, the system determines the emitter’s Z-position without the need for Z-scanning. This method, combined with experimental point spread function calibration, offers a powerful and robust approach to 3D localization.
In bypass imaging mode, mirrors adjust to provide a larger field of view, enhancing the versatility of the system. This capability is crucial for applications requiring detailed examination of tissue sections, organoids, or cell colonies.
Top-Hat Illumination: Ensuring Consistent Imaging Resolution
Top-hat illumination is critical for maintaining consistent imaging resolution across the entire field of view. The Vutara VXL employs five powerful laser lines to achieve bright, even illumination, supporting fast imaging speeds. This uniformity is essential for ensuring that resolution remains consistent, even at the edges of the field.
The system’s design contrasts with traditional Gaussian illumination, which can result in reduced resolution toward the field’s periphery. The Vutara VXL’s top-hat profile ensures that localization precision is maintained throughout the imaging process.
Silicon Immersion Objectives: Ideal for Deep Imaging
Silicon immersion objectives offer significant advantages for deep imaging, maintaining consistent quality throughout the sample’s depth. The Vutara VXL comes equipped with the Olympus 60X silicon immersion objective, which provides a higher numerical aperture and a more consistent point spread function (PSF) as imaging progresses deeper into the sample.
This consistency is vital for achieving bright, clear images, making silicon immersion objectives the preferred choice over traditional oil or water immersion objectives, which can suffer from PSF degradation or index mismatches.
Supporting Multiplexed Imaging Workflows
The Vutara VXL is particularly well-suited for multiplexed imaging workflows like DNA-PAINT. This technique relies on the transient binding of dye-labeled DNA strands to generate single-molecule signals, allowing researchers to conduct high-throughput experiments with ease.
DNA-PAINT’s inherent multiplexing capability is supported by the Vutara VXL’s integrated fluidics and software through the PlexFlo platform. With 15 fluidic reservoirs, PlexFlo allows for the simultaneous use of multiple probes, facilitating complex workflows and ensuring that metadata is accurately tracked throughout the imaging process.
Expert Insights and Future Implications
According to Winfried Wiegraebe, Bruker’s product manager for super-resolution microscopy, and Samuel Penwell, a technology development systems engineer, the Vutara VXL represents a significant advancement in microscopy technology. Their combined expertise highlights the system’s potential to transform research methodologies, offering new possibilities for detailed, multiplexed analysis.
As the field of microscopy continues to evolve, the Vutara VXL stands as a testament to the power of innovation. By addressing the limitations of traditional imaging techniques and offering a versatile, high-performance solution, Bruker is paving the way for new discoveries in molecular biology and beyond.
With its robust design, advanced capabilities, and support for complex workflows, the Vutara VXL is poised to become an indispensable tool for researchers worldwide, driving forward the frontiers of scientific exploration.
