SPCImage

Analysis Software for FLIM, PLIM
  • Analysis of Single-Decay, FLIM and PLIM Data
  • Analysis by Iterative Convolution and Fit Procedure or by First Moment of Photon Distribution
  • Calculations of Decay Parameters and Lifetimes by Various Decay Models
  • Calculation of FRET Efficiencies
  • Display of Lifetime and FRET Images
  • Display of Lifetimes, Amplitudes, Intensities or Ratios
  • Export of Lifetime Data and Images

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The ‘SPCImage’ software is used for fluorescence decay analysis. SPCImage can be used both for TCSPC Imaging (FLIM and PLIM) data, and for single-curve fluorescence decay analysis. The SPCImage software is integrated in the bh TCSPC software package. There is a direct link from the SPCM software to SPCImage.

SPCImage uses an iterative convolution procedure. It fits the decay data by single-, double- and triple exponential decay models. An ‘incomplete-decay’ model is available to account for fluorescence left from previous excitation pulses. In FLIM and PLIM data, SPCImage runs the fit procedure in the individual pixels of the image. The lifetime of a single-exponential decay function, amplitude or intensity-weighted lifetimes of multi-exponential decay functions, lifetime components and amplitudes of the decay components, fractional intensity contributions, or ratios of amplitudes, lifetimes and intensity contributions are displayed as colour of the image. Thus, images of local environment parameters such as pH, oxygen or ion concentrations, of the FRET efficiency, of the distance and fraction of interacting proteins, or bound and unbound fluorophores can be produced.

Since version 6.0 SPCImage FLIM analysis software combines time-domain multi-exponential decay analysis with the phasor plot. In the phasor plot, the decay data in the individual pixels are expressed as phase and amplitude values in a polar diagram. Independently of their location in the image, pixels with similar decay signature form clusters in the phasor plot. Different phasor clusters can be selected, and the corresponding pixels back-annotated in the time-domain FLIM images. The decay functions of the pixels within the selected phasor range can be combined into a single decay curve of high photon number. This curve can be analysed at high accuracy, revealing decay components that are not visible by normal pixel-by pixel analysis. SPCImage can also be used to analyse single decay curves, such as data from cuvette experi-ments.

  • Deconvolution of Single-Curve, FLIM, PLIM, FLITS, Multi-Wavelength FLIM and Mosaic FLIM Data
  • Pseudo-Global Analysis FLIM Analysis
  • Batch Processing
  • Single-, Double- and Triple Exponential Models
  • Incomplete-Decay Model
  • Shifted-Component Model
  • First-Moment Analysis
  • Automatic IRF or Measured IRF
  • Binning, Overlapping Rectangular or Circular
  • Display of Single- and Multi-Exponential Decay Data
  • Amplitude- or Intensity-Weighted Lifetimes
  • Lifetimes and Amplitudes of Components
  • Intensity Contributions of Components
  • Quotients of Lifetimes or Amplitudes
  • FRET Efficiencies
  • Fluorescence Lifetime Redox ratio (FLIRR)
  • ROI Definition
  • One- or Two-Dimensional Parameter Histogramms
  • Phasor Plot, Combined with Time-Domain Analysis
  • Image Segmentation by Phasor Plot
  • Parameter Histogramms of Segmented Areas
  • Combination of Pixels Selected by Phasor Plot
  • Precision Time-Domain Analysis of Combined Pixel Data

For more details please see:

  • bh TCSPC handbook, chapter SPCImage Data Analysis Software
  • DCS handbook, chapter SPCImage Data Analysis Software
  • Zeiss handbook, chapter SPCImage Data Analysis Software

The bh TCSPC Handbook
7th edition, corrected and updated September 2017

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Current Version: SPCImage 7.4

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SPCImage Requires a Valid Licence Key to Run. Enquire Now!

 

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