PicoQuant: SymPhoTime64

The SymPhoTime 64 software package is an integrated solution for data acquisition and analysis using PicoQuant's time-resolved confocal microscope MicroTime 200, LSM upgrade kits or TCSPC electronics. Its clearly structured layout and powerful analysis routines allows the user to focus on the results rather than on the data processing. The software is designed for a 64 bit operation system and features a graphical user interface (GUI), which guides the user through all necessary steps for an individual analysis or measurement process. Data dependencies are directly visible in the underlying workspace concept. An integrated scripting language („STUPSLANG“) even puts the user in a position to freely add new analysis procedures or customize existing ones.

The SymPhoTime 64 is the dedicated data acquisition software for PicoQuant's time-resolved confocal microscope MicroTime 200 and LSM upgrade kits. It can of course also be used with custom setups based on PicoQuant TCSPC electronics. All data acquisition and analysis features of the SymPhoTime 64 are based on the unique time-tagging modes of the PicoHarp 300 or HydraHarp 400. In these modes, photons on each detection channel are tagged with the absolute arrival time since the beginning of the measurement and, in certain cases, also along with the time difference to the last laser pulse. This scheme preserves all photon timing information and allows a large range of data interpretation ranging from simple TCSPC histograms to complex imaging and correlation analysis.

With the SymPhoTime 64 analysis of (time-resolved) imaging measurements will be easier than ever before. The software provides special adapted interfaces for many standard analysis procedures ranging from Fluorescence Lifetime Imaging (FLIM) to Fluorescence Resonance Energy Transfer (FRET) and Anisotropy. Each interface only makes those procedures available that are directly required for the individual analysis. This ensures a steep learning curve as well as quick and correct analysis results.

The SymPhoTime 64 also sets a new standard for analysis of fluorescence correlation spectroscopy measurements. The software provides a wide range of specially adapted correlation analysis procedures, which range from classical auto-correlation (FCS) and cross-correlation (FCCS) to lifetime based correlation analysis (FLCS) and total correlation. Even coincidence-correlation analysis is possible based on the unique time-tagging modes of the PicoQuant TCSPC modules. By exploiting the full power of a multi-core computer system, the SymPhoTime 64 is one of the fastest software correlators on the market.

The analysis of fluorescence intensity time traces is another core feature of the SymPhoTime 64. Fluorescence intensity time traces display the measured fluorescence dynamics and can be analysed in a variety of ways. Prominent examples are classical single molecule methods such as on/off histograms, burst size histograms or fluorescence lifetime traces.

The software is available in three different packages in order to meet the different needs of the individual users:

	SPT64 1A	SPT64 1	SPT64 2	SPT64 1+2
Data acquisition features
Direct data acquisition in TTTR mode using PicoHarp 300 or HydraHarp 400
Control of Physik Instrumente scanner (E-710) with scan table of 100 x 100 x 100 µm scan range
Support for Laser Scanning Microscopes (LSM) and generic scanners via external trigger signals
Router support (separation of up to eight detector signals)
Multi point measurements
Data analysis features
Time gating for all methods
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
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
Fluorescence Correlation Spectroscopy (FCS) incl. Pulsed-Interleaved Excitation (PIE) treatment, (auto- or cross correlation) & FCS fitting with bootstrap error analysis
Fluorescence Lifetime Correlation Spectroscopy (FLCS)
FCS calculation and time trace display during the measurement ("online time traces", "online-FCS")
Total correlation
Förster Resonance Energy Transfer (FRET) incl. Pulsed-Interleaved Excitation (PIE) treatment, corrections for direct excitation and bleedthough
Multi-channel scaling analysis - Lifetime and intensity time traces
Burst integrated fluorescence lifetime (BIFL) analysis
On / Off time histogramming and analysis
Burst size histogramming
Calculation of Photon Counting Histogramm usable for PCH
Generic ASCII export filter
Scripting ("STUPSLANG")