LSM Upgrade Kit

The excitation module provides flexible, multi-color picosecond laser illumination from 405 to 900 nm. Multiple laser heads are combined in a compact Laser Combining Unit (LCU) for single- or multicolor operation and fiber-coupled into the LSM for low-background excitation. Repetition rate and laser power can be adapted to match fluorescence or phosphorescence lifetimes while minimizing bleaching. Advanced excitation schemes, including pulsed interleaving, pulse trains for long-lived probes, and automated wavelength switching, are fully supported. Multi-photon excitation can be integrated when required.

The TCSPC module provides precise timing down to the picosecond range while covering long-lived emission processes up to the millisecond regime. This broad dynamic range enables accurate FLIM, PLIM, FRET, and FCS measurements within a single acquisition workflow. All photons are recorded in a Time-Tagged Time-Resolved (TTTR) format, capturing arrival time, detection channel, and scanner synchronization signals. This universal data format supports high-throughput imaging, correlation analysis, and advanced time-resolved experiments with minimal acquisition time.

The detection module offers single-photon sensitivity and supports up to four parallel channels for multicolor, polarization-resolved, and deep-tissue measurements. Configurable for confocal and non-descanned detection, it enables flexible setups for FLIM, FRET, FCS, anisotropy, and correlation analysis. Confocal configurations use fiber coupling for precise spatial detection, while non-descanned pathways support multiphoton excitation and high-efficiency collection in scattering samples. This versatility allows accurate lifetime separation, improved signal quality, and reliable measurements across life science and materials research.

SymPhoTime 64 provides intuitive data acquisition and online analysis for FLIM, FRET, FCS, FLCS, anisotropy, and related time-resolved methods. A dedicated interface to the LSM enables fast previews, large-format FLIM imaging, time series, z-stacks, and multipoint measurements within a transparent, TTTR-based workflow.

For advanced offline analysis, NovaFLIM delivers high-performance, GPU-accelerated processing of FLIM, FLIM-FRET, and anisotropy data. Powerful batch analysis of z-stacks, time-lapse series, and stitched images, combined with flexible and reproducible ROI handling, ensures efficient, publication-ready quantitative imaging.

Using the LSM Upgrade Kit on an Olympus FluoView FV1000, FLIM enables quantitative mapping of resting Ca²⁺ concentrations in neurons and astroglia. The lifetime of Oregon Green BAPTA-1 correlates directly with nanomolar Ca²⁺ levels and remains unaffected by intensity fluctuations, photobleaching, pH, viscosity, or ion concentration changes.

Fluorescence anisotropy imaging of C6-NBD-PC–labeled CHO cells reveals differences between liquid-ordered and liquid-disordered membrane phases. Higher anisotropy in horizontal membranes indicates restricted probe rotation and ordered lipid organization. Measurements were performed on an Olympus FluoView FV1000 equipped with the LSM Upgrade Kit and polarization extension.

The LSM Upgrade Kit enables precise Fluorescence Correlation Spectroscopy (FCS) measurements within defined cellular subcompartments. Diffusion analysis of GFP-Ago2 in nucleus and cytoplasm of ER293 cells revealed compartment-specific mobility differences, reflecting variations in RISC complex size and assembly state.