FLIM Systems for Zeiss LSM 710 / 780 / 880 / 980

Description

The FLIM systems for the Zeiss LSM 710 /780 / 880 / 980 family laser scanning microscopes are based on bh’s Multi-Dimensional TCSPC technique and 64 bit Megapixel FLIM technology. The systems feature single-photon sensitivity, excellent spatial and temporal resolution, multi-exponential decay analysis, and short acquisition time. The systems are compatible both with multiphoton excitation and diode laser excitation. The system fully exploits the fast beam scanning capability of the Zeiss LSM. The functions include advanced techniques like multi-wavelength detection, Z stack FLIM, time-series FLIM, spatial and temporal mosaic FLIM, fluorescence lifetime-transient scanning (FLITS), and phosphorescence lifetime imaging (PLIM). Main applications are the measurement of molecular environment parameters, FRET measurements, autofluorescence FLIM of tissue, metabolic imaging, and plant physiology. For details please see 350-page user handbook, 'Modular FLIM systems for Zeiss LSM 710/780/880 Laser Scanning microscopes' and Addendum to Handbook for LSM 980 Microscopes.

Specifications

Selected Specifications

Lifetime measurement: time-domain
Excitation: High-frequency pulsed lasers
Principle: TCSPC FLIM by bh's Multi-dimensional TCSPC technique
Buildup of lifetime images: Distribution of photons over arrival times and scan coordinates
Scan rate: Works at any scan rate
Buildup of fluorescence correlation data: Correlation of absolute photon times
General operation modes: FLIM in two spectral or polarisation channels, Multi-wavelength FLIM
Time-series FLIM, Microscope-controlled time series, Z-Stack FLIM, Mosaic FLIM, x,y, z, temporal,
Excitation-wavelength multiplexed FLIM, FLITS (fluorescence lifetime-transient scanning), PLIM (phosphorescence lifetime imaging) simultaneously with FLIM, FCS, cross FCS, gated FCS, PCH,
single-point fluorescence decay recording, single-point phosphorescence decay recording

TCSPC System: bh Simple Tau  or Power Tau TCSPC system
Number of parallel TCSPC / FLIM channels: typ. 2, max 4
TCSPC / FLIM modules: SPC-150 NX or SPC-180 NX
Electrical time resolution: 1.5 ps rms / 3.5 ps fwhm
Minimum time channel width: 405 fs
Timing stability over 100 seconds: Better than 0.8 ps rms
Timing stability over 30 minutes: Better than 5 ps rms
Saturated count rate: 10 MHz per channel
Input from detector: constant-fraction discriminator
Reference (SYNC) input from Laser: constant-fraction discriminator
Synchronisation with scanning: via frame clock, line clock and pixel clock pulses
Scan rate: works with any scan rate
Synchronisation with laser multiplexing: via routing function of TCSPC modules
Recording of multi-wavelength data: simultaneous in 16 channels, via routing function
Experiment trigger function: TTL, used for Z stack FLIM and microscope-controlled time series
Operation modes of TCSPC system: Single f(t), oscilloscope, f(txy), f(t,T), f(t) continuous flow
FIFO (correlation / FCS / MCS) mode, Scan Sync In imaging, Scan Sync In with continuous flow
FIFO imaging, FIFO Imaging combined with with MCS imaging, mosaic imaging, time-series imaging
multi-detector operation, laser multiplexing operation, cycle and repeat function, autosave function
Display Functions (online): Intensity images, gated intensity images, lifetime images, decay curves in ROIs, decay curves, FCS curves, intensity traces.
No. of images displayed simultaneously: 8
Max. Image Format, pixelsX x pixels Y x time channels: 4096 x 4096 x 64, 2048 x 2048 x 256, 1024 x 1024 x 1024, 512 x 512 x 4096, 256 x 256 x 4096

Software:
Data Acquisition Software: bh SPCM
Operating System: Windows 10 64 bit
Data Analysis Software: bh SPCImage NG
Principle of Data Analysis: MLE with GPU processing
Model functions: Single, double, triple exponential decay, single, double, triple exponential decay with incomplete decay, shifted component model.
IRF modelling: Syntethic IRF function fit to decay data

Excitation Sources, for confocal FLIM: One to four ps diode lasers
Available wavelengths: 375 mm to 785 nm
Repetition rate: 20, 50, 80 MHz and CW
Pulse width: 40 ps to 100 ps
Excitation source, for multiphoton FLIM: Ti:Sa laser of Zeiss LSM NLO system

Detectors: HPM-100-40 hybrid detector with GaAsP cathode, 300 to 710 nm
Optional: HPM-100-06 detector with <20 ps fwhm IRF width, 300 to 600 nm
Optional: HPM-100-50 detector, 400 to 900 nm
Optional: MW-FLIM GaAsP multiwavelength detector

For more specifications please see FLIM Systems for Zeiss LSM 710 / 780 / 880 Family Microscopes, user handbook.

Documents

The realm of the bh FLIM systems are in molecular imaging. Typical applications are the imaging of ion concentrations, pH, or local viscosity, protein interaction experiments by FRET, and metabolic imaging by fluorescence decay of NADH an FAD in combination with oxygen measurement. In these applications, the bh FLIM systems benefit from their high sensitivity, high time resolution, high timing stability, and their capability to resolve multi-exponential-decay profiles into their components. Other advantages are the capability to record FLIM of fast physiological effects down to the millisecond range, and to record at several excitation and emission wavelengths simultaneously.

Gallery

• Multiphoton FLIM
• Confocal FLIM
• Parallel Recording in 1 to 4 Wavelength Channels
• Multiplexed Recording at two Excitation Wavelengths
• Multi-Wavelength FLIM in 16 Wavelength Channels
• Multiphoton Multi-Spectral NDD FLIM
• Fast Acquisition FLIM by Optional FASTAC FLIM System
• Near-Infrared FLIM
• Time Series FLIM: Fast time Series or Microscope-Controlled Time Series
• Z Stack FLIM
• Spatial Mosaic FLIM
• Temporal Mosaic FLIM of Physiological Effects, down to 40ms per Image
• FLITS: Recording Transient Lifetime Effects in a Line Scan, Resolution down to 1 ms
• PLIM (Phosphorescence Lifetime Imaging) Simultaneously with FLIM
• Fast Preview Mode
• Easy Switching Between Instrument Configurations and Operation Modes
• Runs at Any Scan Rate
• High Resolution Images, Up to 2048 x 2048 Pixels
• FCS and Cross-FCS, No afterpulsing Due to Use of Hybrid Detectors