Openlab. Modular imaging software for cell biologists


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Openlab Application – Ratio Imaging

The Openlab Ratioing module provides rapid ratioing of image sequences of extended dynamic range, up to 16 bit monochrome data. Maximum and minimum ratio display values and threshold settings can be set for each source image. Look Up Tables and Intensity Modulated Display can be applied and ratioed images can be created, measured and graphed on- or off-line.

An unlimited number of ROIs can be selected for measurement and the Automator can be set to automatically add event markers to the graph trace for greater accuracy. Ratioed image sequences can be viewed as movies using the Presentation palette and exported as QuickTime movies.

The Automator confers the power to design and perform experiments of unlimited versatility, and allows essential procedures such as background subtraction to be performed automatically.

ratio images

Openlab Application – FRET imaging

The Openlab FRET module supports three image FRET applications. An Openlab imaging system configured for FRET allows you to acquire, create, display, measure and graph FRET result images on- or off-line. It also provides the tools required for background subtraction and bleedthrough calculation.

The FRET module includes documented expressions that can be applied to the input images. The application of the expressions takes place on a pixel by pixel basis to generate the output image. The FRET module dialog provides a preview of the result image and gives you choices for the visual display of the results including linear or logarithmic scale and Intensity Modulation.


FRET donor image	FRET acceptor image	FRET result

Ratio Imaging case study

Professor Andreas Guse and Svenja Kunerth of the University Hospital Hamburg-Eppendorf are studying intracellular Ca^²⁺signaling. In many cell types, intercellular Ca^²⁺ signaling starts with highly localized subcellular events, which are often followed by global Ca^²⁺ signals. Professor Guse is using an Openlab system to establish a technique to allow visualization of subcellular Ca²⁺ signals in living cells using a light microscope acquisition system.

To produce the T-lymphocyte images shown here, a Z series of three planes, 0.5µ apart, was acquired by excitation at 340 and 380nm at each time point. The images were deconvolved using Openlab Nearest Neighbor Deconvolution module, then ratioed and a Rainbow cLUT was applied for display. Using the Openlab Profiling module, it is possible to visualize both the position and intensity of the sucellular Ca²⁺ signals across the cell at each time point.

FRET Imaging case study

Dr Art Alberts at the Van Andel Research Institute is interested in the intracellular signaling networks that regulate cell proliferation, shape and motility.