Microscopy > Software > VistaVision

VistaVision

VistaVision for Alba
VistaVision for Alba is a complete software package for instrument control, data acquisition and data processing. VistaVision enables control of the automated devices on all Alba instruments including shutters, filterwheels, XY stages, and light detectors. A convenient signal monitor displays the signal intensity from each channel in real time, and it is utilized during instrument alignment.

It has been developed for the Windows XP environment in modular components that can be installed independently when constructing a custom-built instrument that uses ISS modular components.

Instrument Control Module
The VistaVision configuration file allows the user to select the proper driver for a variety of components.

Data Acquisition Cards
  • ISS FCS card
  • ISS FastFLIM card
  • B&H SPC-830, SPC-150 cards
Scanning Mirrors Modules
  • ISS scanning mirrors module
  • Cambridge Technology Models
Piezo-controlled Stages
  • ISS XYZ piezo-controlled stage
  • MadCity Models Nanoposition
  • PI Models
Automated Microscopes
  • Nikon Models TE2000-E, Ti-E
Microscopes Stages
  • ASI Model 2000
Laser Launchers
  • ISS Laser Launchers Series

Imaging Module
The imaging module of VistaVision includes the following components:

  • Imaging (steady-state)
  • FLIM digital frequency-domain (FastFLIM)
  • FLIM analog frequency-domain
  • FLIM TCSPC
  • RICS (raster imaging correlation spectroscopy)
  • Polarization

Data can be acquired on one channel or up to four channels simultaneously (depending upon the acquisition modality).

The imaging modules allow for the user to select the image parameters (image sizes, dwell time, z-stack and time series). The software includes operations between images (addition, subtraction, multiplication and division), smoothing, filtering and automatic threshold settings for image contrast enhancement.

FLIM images are analyzed in two modalities:
• Polar plot
• Fitting algorithm (Levenberg-Marquardt minimization routine)

Image series can be exported to the AVI format for movie rendering and to ImageJ and MetaMorph for 3D rendering. Plots are exported to such popular formats as png, jpeg, gif, tiff, bitmap and metafile.

FFS Data Acquisition and Analysis Software
FS data are acquired in either time mode or photon mode. VistaVision features a real-time display of the auto correlation function, G(τ) - apart from a nominal delay (less than one second) required for the computation of the function. A sequence of multiple data acquisition files can be acquired (for instance, when using a microwell plate on a computer-controlled XY stage) and displayed and stored automatically.

VistaVision acquires and stores raw data files, which can be opened at a later time and re-correlated; sections of each raw data file can be purged from the presence of impurities (for instance, when an impurity traverses the observation volume) and re-analyzed. The software also stores the autocorrelation function, the cross-correlation function and the photon-counting histogram data files. All of these files can be exported to several formats.

The acquisition components include the following modules:
• FFS, Fluorescence Fluctuations Spectroscopy
• Scanning FCS (requires the imaging module)

VistaVision includes data analysis routines for using the autocorrelation function, the cross-correlation function, or the photon-counting histogram with different laser-beam point spread functions (one-photon and two-photon). The analysis can be carried out for a single data file, or, using global analysis, for up to 384 data files simultaneously.

The following table summarizes the various analysis options presently implemented in VistaVision, along with the parameters that can be determined using each analysis approach.

Statistical function utilized for FFS data analysis Single set and Global
fitting models available
in the FFS software		 Parameters determined by
the FCS software
Autocorrelation
  • One or two species using:
    • 2D- or 3D-Gaussian PSF
    • 3D-Gaussian-Lorentzian PSF
    • one-photon excitation
    • two-photon excitation
    • presence of flow
  • Up to 384 data sets global fitted.
  • One or two species using:
    • Diffusion coefficient
    • Concentration
    • Triplet state decay time constant
    • Triplet function
    • Flow rate
    • Size of excitation volume
Cross-correlation
Photon Counting Histogram (PCH)
  • One or two species using:
    • 2D- or 3D-Uniform
    • 2D- or 3D-Gaussian PSF
    • 3D-Gaussian-Lorentzian PSF
    • one-photon excitation
    • two-photon excitation
  • Up to 384 data sets global fitted.
  • Number of molecules
  • Molecular brightness
User Defined Equation
  • Up to 50 different user defined equations kept in the panel list for user to choose.
  • Equation could include Sin, Cos, Exponential, etc… Except the integral unclosed form equation.
  • Up to 384 data sets global fitted.
  • Up to 30 parameters allowed in the equation.

Additionally, VistaVision allows for the following analysis modules:
• Fluorescence Lifetime Correlation Spectroscopy (FLCS)
• BIFL

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