All FRET applications are now integrated into the physiology module of the
AxioVision 4.7 microscopy software from Carl
Zeiss. This enables cell and development biologists to use all AxioVision
functionalities such as the enhancement of fluorescence signals and the suppression
of image noise. Operation is very easy as only a few clicks are required.
The FRET software is used to determine the energy transfer portion between
two adjacent protein molecules through fluorescence energy transfer (FRET),
measure the distance between adjacent protein molecules below the microscope
resolution and obtain quantitative temporal and spatial information about the
bonding and interaction between proteins, lipids, enzymes, DNA and RNA in vivo.
The software offers numerous additions to existing and new methods of FRET
measurement by using different corrective techniques or acceptor bleaching.
Users can define their own formulas for the tasks to be performed or use pre-programmed
FRET formulas to make FRET experiments even more simple. Furthermore, it is
possible to modify existing FRET formulas and easily create new evaluation techniques
via the formula field. AxioVision enables all microscope components and software
functions to be used for processing. The dual camera module more than doubles
the speed of capture. Up to 140 frames per second are available with the AxioCam
HS. In addition, all required time series images (donor, acceptor and FRET)
can be captured using the physiology module. However, the system can also be
utilized for all standard tasks in image processing and analysis.
The benefits of the FRET software can be perfectly utilized with the motorized
Axio Imager or Cell Observer microscope systems. However, suitable filter sets
for FRET measurements also allow the software to be used with manual fluorescence