Multispectral High-Resolution Fluorescence Imaging System (Fast Spinning Disk Two Cameras Nikon System)

Medical University of Graz

Graz | Website

Open for Collaboration

Short Description

Application Areas:
The Multi-Spectral High-Resolution Fluorescence Imaging System (Fast Spinning Disk Two Cameras Nikon System) is particularly well-suited for visualizing rapid processes such as ion changes in individual living cells and their subcellular structures, including cell organelles like mitochondria, through fluorescence microscopy. Furthermore, the system can effectively be utilized for multiplexing applications, simultaneously capturing multiple fluorophores with different spectral properties (> 4). It is also suitable for measuring Förster Resonance Energy Transfer (FRET)-based biosensors, such as CYF-YFP and GFP-RFP FRET pair-based biosensors, as well as single fluorescence proteins (FP)-based biosensors with various colors (blue, green, yellow, orange, red).

Technical System Description:
The system is built upon a Nikon Ti2 inverted microscope with specialized optics for a large field of view of 35 mm. It comprises a wide-field epi-fluorescence section with an LED light source (CoolLED pE800 MB) emitting wavelengths of 400 nm, 435 nm, 470 nm, 500 nm, 550 nm, 580 nm, 625 nm, and 740 nm. Additionally, it features a confocal section with a rapid spinning disk array unit (Crest X-Light V3) and 7 laser lines (Lumencor Celesta) emitting wavelengths of 405 nm, 446 nm, 477 nm, 520 nm, 546 nm, 638 nm, and 749 nm. At present, the system is equipped with a 100x oil objective, a 40x water objective, and a 20x air objective. The system is equipped with two fast sCMOS cameras (Photometrics Kinetix; 3200 x 3200, 6.5 µm pixel size; QE > 95%, F-Mount; 498 fps at 8-bit, 83 fps at 16-bit),

Additional Equipment:
The system also includes a gravity-driven 9-channel perfusion system (NGFI GmbH), a vacuum pump, and a stage incubator for temperature and CO2 control, enabling the creation of incubator conditions directly at the microscope.

Contact Person

Roland Malli

Research Services

Access via core facility or scientific unit.

Methods & Expertise for Research Infrastructure

Our research infrastructure integrates biomedical research expertise with the Multi-Spectral High-Resolution Fluorescence Imaging System (Fast Spinning Disk Two Cameras Nikon System). This system enables the investigation of rapid processes such as ion changes in individual living cells and their subcellular structures, including organelles like mitochondria. Leveraging our longstanding experience in developing and applying biosensors, we effectively utilize the capabilities of this system. Specifically, we specialize in multiplexing applications, allowing for the simultaneous capture of multiple fluorophores with diverse spectral properties.

Additionally, we successfully employ Förster Resonance Energy Transfer (FRET)-based biosensors and single fluorescence protein (FP)-based biosensors with various colors. Our utilization of this advanced research infrastructure and our robust expertise have contributed to significant insights into biomedical and natural sciences research and hold the potential for developing innovative approaches to disease diagnosis and treatment.