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LIFA-X, for long lifetime phosphorescence imaging

 
 

the newest addition to the LIFA product line is the LIFA-X. LIFA stands for Lambert Instruments Fluorescence lifetime imaging Attachment. The LIFA-X is for long lifetime imaging microscopy of lifetimes from 0.1 ns up to 1 ms (phosphorescence). The ‘X represents the extended range of lifetimes that can be resolved when compared to the LIFA (0.1 ns ? 300 ns). The LIFA-X can be used for, e.g., measuring oxygen concentrations using optodes or reactive oxygen species in cells.

The LIFA-X is based on the LIFA (Lambert Instruments Fluorescence lifetime imaging Attachment) for FLIM -Fluorescence Lifetime Imaging Microscopy and is designed in such a way that it can be retro fitted onto an existing LIFA system. Therefore, it has the same basic features and advantages as the lambert Instruments

 
LIFA-X, for long lifetime
(phosphorescence) imaging
 


Fluorescence lifetime imaging?Attachment?:

  • it can be attached to any widefield fluorescence microscope
  • works in the homodyne frequency domain
  • is easy to install (within 1 hour) and to operate>
  • does not require an optical bench with laser.
The LIFA system has been judged "easy and highly quantitative". Please read our white paper for more technical information on the LIFA-X

Components

For excitation typically modulated LEDs are supplied, however, modulated laser diodes are optional when higher intensity of excitation is essential (ie.g., for spinning disk confocal or TIRF - Total Internal Reflection Fluorescence).

Image Intensifier Tube

As for the LIFA -Lambert Instruments Fluorescence Attachment, the LIFA-X can be supplied with different image intensifier tubes. For GenII (Gen 2) we normally offer either the S20 or the SuperS25 photocathode. For higher quantum efficiency (25% instead of 10%) the GenIII (Gen 3) image intensifier is offered, with the GaAs photocathode. See here for the photocathode quantum efficiency characteristics:
 
       
 


Digital CCD camera

The image intensifier (tube) is fiber-optically coupled to a CCD camera and integrated in the LI2CAM MDX camera that is made by Lambert Instruments. The intensified CCD camera LI2CAM MDX has larger dimensions than the LI2CAM, intensified CCD camera that is supplied with the LIFA because of the integrated pulse generator. The pulse generator is activated for low frequency modulation. Additionally, a separate pulse amplifier is delivered for converting the LI2CAM MDX generated pulses to signals suitable for the modulated LED light source.

Integrated Control unit / Signal generator / Power supply

This unit supplies the RF modulation signals at controlled frequency (up to 120MHz), amplitude and phase difference to the modulated LED or laser light source and to the image intensifier. The control unit also supplies the DC power and high voltage to the modulated LED and image intensifier. An extra output to drive an external modulator is available, as well as monitor outputs to check the modulation signals.

LI-FLIM software

The user-friendly LI-FLIM software package is developed by Lambert Instruments. With this package you can acquire single- and multifrequency lifetime images, process them and do analyses by statistics or polar plot. Time series can be done as well, for which the frequency domain FLIM –Fluorescence Lifetime Imaging Microscopy is highly appropriate because of its fast acquisition possibilities. The LI-FLIM software automatically recognizes connected devices. Among others different light sources are integrated in LI-FLIM like the modulated LEDs, directly modulated laser diodes, and the Cobolt DPSS laser (not modulated). Moreover the nanopositioning system NanoF100 (z-stepper) is integrated in the LI-FLIM software which is ideal when using the LIFA with the Yokogawa spinning disk. Customers receive the password for the newest LI-FLIM software release which can be downloaded from the LI-FLIM software page directly.
 
         
  Patents
Patents owned by Photonic Research Systems Ltd.