Becker & Hickl have upgraded their FLIM systems for the Zeiss LSM 980 laser scanning microscopes. Standard systems have two parallel ultra-fast recording channels, additional channels can be added on demand. Using the new SPC-180 NX TCSPC FLIM modules and ultra-fast hybrid detectors, an IRF width of less than 20 ps FWHM (8 ps RMS) is obtained. Systems are available both for multiphoton and for confocal microscopes. Multiphoton versions use non-descanned detection to obtain clear images from deep layers of biological tissue. Confocal systems have the detectors attached to an optical output from the scanner, using the full range of filter- and pinhole-selection features of the LSM 980. Detectors for both versions are compatible and can be swapped between the output ports of the microscope. Alternatively, pairs of detectors can be attached to both the confocal port and the NDD port, and one or the other be activated by software command. The new bh / Zeiss FLIM systems are available with bh’s new integrated SPCConnect software. It combines bh SPCM software and Zeiss ZEN software by TCP (Transmission Control Protocol). That means SPCM is running together with ZEN, with both software components freely exchanging commands, system parameters, and data. Compared to an integration on the DLL level TCP integration has the advantage that, in addition to basic FLIM, functions like Z-stack recording, time-series recording, fast triggered accumulation, FLITS, and simultaneous FLIM / PLIM are available. Upgrades on both sides become immediately available to the user. Data analysis is performed by bh’s legendary SPCImage data analysis software. It performs single, double and triple decay analysis and directly delivers lifetimes, component amplitudes, and biologically relevant parameters such as metabolic ratios, FLIRR, FRET efficiencies, and FRET distances. SPCImage runs on a GPU, delivering FLIM results within seconds even for image sizes in the megapixel range. With these features, the new FLIM systems are excellently suitable for the whole range from entry-level FLIM to high-end multidimensional molecular imaging applications.
