FLIM was recorded on a Zeiss LSM-780 NLO confocal/multiphoton microscopy system comprising an inverted Axio Observer Z1 microscope, an X-Cite 120PC Q mercury arc light source (Excelitas Technologies) for cell selection, a motorized stage for automated scanning, an IR Chameleon Vision-II ultrafast Ti:sapphire laser for multiphoton excitation (Coherent), a Zeiss 40 × 1.3 NA oil immersion Planapo objective, an environmental chamber (PeCon GmbH, Germany) that envelops the microscope stage to control temperature and CO2 level, and a 3-channel FLIM system based on three HPM-100–40 GaAsP-based hybrid detectors and 3 SPC-150 TCSPC boards (Becker & Hickl). The SPC-150 boards are synchronized with the 2-photon excitation laser and the Zeiss LSM-780 NLO scan head signal. Ex 740 nm; Em450/50 nm.
Spc 150 tcspc boards
Manufactured by Becker & Hickl
Sourced in Germany
The SPC-150 TCSPC boards by Becker & Hickl are time-correlated single-photon counting (TCSPC) modules designed for time-resolved fluorescence and luminescence measurements. The boards provide high-resolution timing capabilities and can be used in a wide range of applications.
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Lab products found in correlation
3 protocols using spc 150 tcspc boards
1
Multiphoton Microscopy Imaging Setup
2
Fluorescence Lifetime Imaging of FMR-1 and FMR-2
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Cells were imaged in 8-well plates on a inverted TCS-SP2 microscope (Leica Microsystems, Germany) and SPC-150 TCSPC boards (Becker & Hickl, Germany). FMR-1 and FMR-2 were excited using a picosecond-pulsed (90 ps optical pulse width, 20 MHz repetition rate) 467 nm diode laser (Hamamatsu, Japan) through a 63X 1.2 NA water objective, with a 485 nm dichroic mirror and a 500 long pass filter. All imaging was done at room temperature. Each FLIM image was acquired over 900 s to ensure as little noise as possible (much shorter acquisition rates are possible). Lifetime histograms and FLIM images were analysed using SPCImage software (Becker & Hickl, Germany).
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Multiphoton Microscopy Imaging Setup
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