LA Software

FRAP

_{From left to right: Yeast cells expressing GFP before bleaching, during bleaching and after recovery; images obtained with an iMIC, fitted with Yanus Scan Head, Polytrope Imaging-Mode Switch, Dichrotome Dual Emission Extension, 60x Objective, Sensicam QE;
Images courtesy of Dr. Roland Wedlich and Felix Spira, MPI Martinsried;}

FRAP recording

TILL's Live Acquisition enables photo bleaching of multiple, freely definable regions in a specimen. A FRAP experiment can be recorded in widefield or TIRF illumination mode and either can be combined with the FRET mode in an iMIC. This makes it possible to record two emission wavelengths simultaneously, saving time and treating the cells with care.
Switching from FRAP to widefield or TIRF is done within milliseconds thanks to the innovative Polytrope Image Mode switch. This ensures that the first image after the end of the photo bleaching is immediately acquired.

Features

• Cost effective solution in TIRF and Widefield applications
• Multiple user defined regions (spot, line, rectangle, ellipse, polygon)
• ultra fast switching between FRAP and image acquisition
• Multi-Channel image acquisition (TIRF or Widefield)

FRAP evaluation

TILL's LA FRAP module provides evaluation possibilities of FRAP experiments. This is done by calculating the mean intensities of several ROIs. For sophisticated evaluation and calculation of mobile and immobile fractions, the LA FRAP module uses ROIs for Whole cell, Background and Bleaching correction in addition to the actual FRAP ROI. All ROI mean intensities in a FRAP experiment are displayed in a graph. An additional graph provides normalized values of the FRAP experiment after the FRAP time data point. With LA FRAP it is also possible to calculate the corresponding function, single and double exponential. For further evaluation the calculated data can be exported and the graphs can be saved to a file.

Features

• ROIs for: Whole chell, Refernce and Bleaching correction
• Normalized graph with two fitting functions (single and double exponential)