FLIMfit

FLIMfit is a software package for the analysis and visualisation of time-resolved data from FLIM (Fluorescence Lifetime Imaging) measurements. It uses highly efficient algorithms that can globally fit FLIM data with modest photon numbers to complex decay models, across hundreds or thousands of fields of view, requiring only tens of seconds of processing time on a reasonable desktop computer. This makes it useful to analyse time series or multiwell plate FLIM data. The algorithms used in FLIMfit are discussed in detail in Warren et al, 2013.

Contents

• FLIMfit
• GUI Documentation
  • Menus
  • Fitting options
  • Instrument and preprocessing options
• Loading Plate Data in FLIMfit
  • Setup calibration data
  • Loading data
• Advanced Topics
  • Creating a library IRF
  • Determining a reference lifetime
• Experimental design
  • General Considerations
  • Multi-well plate experiment design
  • Choosing a reference dye
• Multiphoton TCSPC FLIM Checklist
  • Before imaging
  • During imaging
  • After imaging
• Time Gated FLIM Checklist
  • Setting up imaging
  • Before imaging
  • During imaging
  • After imaging
• Handling time varying backgrounds
• Frequently asked questions
  • Fitting
  • Exporting Data
  • New Users
  • Estimating IRF shift
  • Estimating the G factor

Common Abbreviations

FLIM	Fluorescence Lifetime Imaging Microscopy
TCSPC	Time Correlated Single Photon Counting
IRF	Instrument Response Function
TVB	Time Varying Background
FOV	Field of View (synonymous to “image”)

Indices and tables

• Index
• Module Index
• Search Page