SymPhoTime 64

Fluorescence Lifetime Imaging and Correlation Software

Software for comprehensive time-resolved fluorescence analysis and imaging.
SymphoTime 64 software interface for fluorescence lifetime imaging analysis

Key Benefits

Faster insights from complex data
Efficient imaging and analysis workflows
High accuracy in lifetime fitting
Reliable correlation across time scales
Customizable tools for unique experiments
Seamless control of microscopy hardware

Software for Advanced Fluorescence Experiments

Integrated Analysis Workflows

SymPhoTime 64 is an integrated software platform for advanced time-resolved fluorescence imaging and analysis. Designed to meet the demands of diverse research fields, it combines clear, structured workflows with powerful acquisition and processing tools. The software supports a broad range of fluorescence techniques and offers customizable analysis procedures that let users focus on scientific insight rather than data handling. Whether working with complex imaging, correlation spectroscopy, or single-molecule experiments, SymPhoTime 64 provides a flexible, future-ready environment for extracting accurate and meaningful results from time-resolved measurements.

SymPhoTime 64 combines imaging, correlation, and time-trace analysis in a single software environment, enabling flexible evaluation of time-resolved fluorescence data across diverse experimental workflows.

Advanced Tools for Imaging, Correlation, and Dynamics

SymPhoTime 64 unifies advanced imaging, correlation, and time-trace analysis within one flexible software environment. It supports FLIM, FRET, anisotropy, and multichannel imaging, complemented by accurate decay fitting and IRF reconvolution. Its high-performance correlation engine enables FCS, FCCS, FLCS, antibunching, and correlation workflows with precise parameter extraction. Dedicated interfaces further streamline single-molecule studies, including blinking, burst analysis, lifetime traces, and PIE-FRET. Together, these capabilities provide a powerful platform for resolving dynamic processes across diverse fluorescence experiments.

Example of the dark theme GUI for low light measurements.

Flexible Control and Custom Configuration Options

SymPhoTime 64 combines integrated data acquisition, flexible hardware control, and extensive customization in one adaptable platform. It supports PicoQuant microscope systems, LSM Upgrade Kit, time tagging & TCSPC Electronics, and custom setups, offering synchronized multi-mode imaging and real-time previews for immediate quality assessment. Through the STUPSLANG scripting language, users can create tailored analysis routines, fitting models, and hardware extensions. This seamless compatibility with photon counting instrumentation and modular fluorescence systems enables precise, application-specific configurations for advanced research.

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Matching Applications & Methods​

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Specifications

Supported TCSPC modulesHydraHarp 500, PicoHarp 330, TimeHarp 260, MultiHarp 150, MultiHarp 160
Supported configurationsMicroTime 200 with 100 x 100 (x 100) µm or 10x7.5 cm piezo scanner or FLIMbee galvo scanner MicroTime 100 Laser Scanning Microscopes (LSM)from Nikon, Olympus or Zeiss Stand-alone TCSPC modules Remote control via TCP/IP interface (software handshake with ZEN and NIS Elements)
Routing1 to 8 detectors
Measurement modesSingle point, multi-point, 2D imaging (XY, XZ, YZ), 3D imaging (XYZ), time lapse (XYT), oscilloscope mode for alignment purposes
Measurement previewsFLIM, FCS, FLCS and FCCS, time trace, TCSPC histogram, parallel calculation and display of up to 4 different previews
Automated measurementsZ–stacks, time ttacks, image stitching, multi-point measurements
Hardware controlPDL 828 "Sepia II" laser driver E-710, E-725, E-727 and wide range scanner controller (Physik Instrumente) Shutter of MicroTime 200 Wide-field fluorescence camera IDS uEye USB3 in MicroTime 200
General featuresTime gating Binning TCSPC binning TCSPC fitting (Multi–Exponential Decay (1 to 5 Exponentials), least–squares fitting, MLE fitting, IRF reconvolution, tailfit, bootstrap error analysis) Global analysis of TCSPC curve fitting GUI themes
ImagingFLIM, FLIM-FRET, intensity FRET, anisotropy imaging, (time-gated) fluorescence intensity imaging Pattern matching, fast pattern matching Adjustable color scale Region-of-Interest (ROI) Bin export for phasor analysis (via third party software globals developed by the Laboratory for Fluorescence Dynamics)
CorrelationFCS, FCCS, FLCS, PIE–FCS STED-FCS, STED-FLCS FCS fitting (models: diffusion constants, triplet state, conformational, protonation, gaussian PSF, user-defined models via scripting, bootstrap error analysis) Global analysis FCS calibration Antibunching/coincidence correlation, total correlation
FRETPIE (pulsed interleaved excitation) Bleedthrough correction FLIM-FRET
STEDSTED, gatedSTED, STED-FLIM, pulsed interleaved STED and confocal, resolution estimation
Fluorescence intensity tracesBlinking (on/off histogramming), count rate histogram (PCH), burst size histogram, intensity–gated TCSPC, fluorescence lifetime traces, lifetime histogram, BIFL (burst integrated analysis)
Steady–state anisotropyObjective correction factors included
Export data formatsBMP, ASCII, TIFF, BIN
User Scripting (STUPSLANG)User–defined analysis procedures, fitting functions, multi–parameter filtering control of external hardware via suited interface
Required PC CPUwith SSE2 and EMT64 or AMD64 extension; recommended: quad–core or better
CPU clock2.2 GHz (or better) quad-core CPU
RAMminimum 4 GB (suggested 16/32 GB)
Operating systemWindows 10 x64
Disk space>= 100 MB (except data storage)
Display(s)Full HD Display
Protection Module (HASP)USB

All Information given here is reliable to our best knowledge. However, no responsibility is assumed for possible inaccuracies or omissions. Specifications and external appearances are subject to change without notice.

Version Features

SPT 64 1A

SPT 64 1

SPT 64 2

SPT 64 1+2

SPT 64 1+2+3

Direct data acquisition in TTTR mode using MultiHarp 150/160, TimeHarp 260, PicoHarp 300, PicoHarp 330 or HydraHarp 400

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Control of Physik Instrumente scanner

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Support for Laser Scanning Microscopes (LSM) and generic scanners via external trigger signals

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STED imaging, STED-FLIM, STED-FCS

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Router support (separation of up to eight detector signals)

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Multi point measurements

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SPT 64 1A

SPT 64 1

SPT 64 2

SPT 64 1+2

SPT 64 1+2+3

Time gating for all methods

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TCSPC-lifetime histogramming and fitting to exponential decay functions up to 5th order incl. tail fitting, reconvolution analysis, maximum likelihood estimation and bootstrap error analysis

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Fluorescence lifetime imaging (FLIM) incl. online-visualisation of FLIM data during the measurement, arbitrary regions of interest for analysis, maximum likelihood estimation and bitmap export

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Pattern matching, fast pattern matching

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Fluorescence correlation spectroscopy (FCS) incl. pulsed-interleaved excitation (PIE) treatment, (auto- or cross correlation) & FCS fitting with bootstrap error analysis

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Fluorescence lifetime correlation spectroscopy (FLCS)

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FCS calculation and time trace display during the measurement ("online time traces", "online-FCS")

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Total correlation

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Förster resonance energy transfer (FRET) incl. pulsed-interleaved excitation (PIE) treatment, corrections for direct excitation and bleedthough

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STED imaging, STED FLIM, gatedSTED, pulsed interleaved STED and confocal, resolution estimation

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Multi-channel scaling analysis – lifetime and intensity time traces

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Burst integrated fluorescence lifetime (BIFL) analysis

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On/off time histogramming and analysis

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Burst size histogramming

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Calculation of photon counting histogramm usable for PCH

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Generic ASCII export filter

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Scripting ("STUPSLANG")

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