Confocal Imaging and Cellular Stimulation with Minimal Phototoxicity

Olympus has released the new FluoView FV1200 confocal laser scanning microscope, optimized for live cell imaging. The new system uniquely combines the accuracy of the newly engineered IX83 frame with enhanced fluorescence sensitivity and simultaneous laser stimulation of cells, making it ideal for advanced life science applications such as FRAP, FLIP and photo-activation. In particular, the new highly-reflective, silver-coated galvanometer scanning mirrors and dual channel GaAsP FluoView PMT module both act to maximize light transfer and detection. This allows for reduced laser power, protecting against the effects of photobleaching and phototoxicity.

The generation of insightful confocal imaging data from living cells depends on optimally balancing laser power and sensitivity to minimize the effects of phototoxicity and photobleaching. To meet this challenge, the FV1200 is designed to maximize sensitivity via a new dual channel GaAsP FluoView high sensitivity PMT module, which delivers up to 45% quantum efficiency. In addition, Peltier cooling reduces electrical noise by 80%, increasing signal to noise ratio.

The new high reflection silver coating on the galvanometer scanning mirrors of the FV1200 ensures durability along with improved reflection efficiency for excitation and emission paths, increasing light efficiency in the visible range by 5 - 15% and IR reflectance up to 22%. This allows for a decrease in laser power, while providing the same level of fluorescence sensitivity. In combination with the unique Olympus SIM scanner, which allows simultaneous illumination and stimulation, it is possible carry out advanced localization experiments such as FRAP and FLIP, without fear of damaging cells.

Accurate confocal imaging also requires the upmost thermal and mechanical stability, as is provided by Olympus’ new fully automated IX83 frame. To further improve accuracy, the Olympus Z-drift Compensation Unit maintains automatic precision focus without damaging cells, and uniquely offers both a ‘one-shot’ and a ‘continuous’ focus mode. In this way, even the longest time-lapse experiments will generate reliable and accurate results.

To maximize application flexibility, the FV1200 combines the GaAsP PMT with the high dynamic range of three standard multi-alkali PMTs. The new GaAsP module is also compatible with the FluoView FV1000, providing an opportunity to easily enhance the performance of the previous system. System operation is controlled via the new FluoView ASW Version 4.0 software, improving direct file access and analysis, and providing a common platform for both the FV1000 and FV1200.

Dr Buelent Peker, Product Manager at Olympus Microscopy Europa, commented: “Given the trend toward longer observation periods using living cells, the new FV1200 system meets the needs of researchers requiring reliable time lapse imaging in combination with highly accurate and sensitive fluorescence observations and simultaneous cellular stimulation. In this way, it enables researchers to obtain reliable data from living cells, while minimizing the risk of causing death or triggering abnormal cellular behavior.”