Fluorescence microscopy has become a widely used tool for the study of medically relevant intra- and intercellular
processes. Extracting meaningful information out of a bulk of acquired images is usually performed during
a separate post-processing task. Thus capturing raw data results in an unnecessary huge number of images,
whereas usually only a few images really show the particular information that is searched for. Here we propose
a novel automated high-content microscope system, which enables experiments to be carried out with only a
minimum of human interaction. It facilitates a huge speed-increase for cell biology research and its applications
compared to the widely performed workflows. Our fluorescence microscopy system can automatically execute
application-dependent data processing algorithms during the actual experiment. They are used for image contrast
enhancement, cell segmentation and/or cell property evaluation. On-the-fly retrieved information is used
to reduce data and concomitantly control the experiment process in real-time. Resulting in a closed loop of
perception and action the system can greatly decrease the amount of stored data on one hand and increases the
relative valuable data content on the other hand. We demonstrate our approach by addressing the problem of
automatically finding cells with a particular combination of labeled receptors and then selectively stimulate them
with antagonists or agonists. The results are then compared against the results of traditional, static systems.
In this paper we present a high-throughput sample screening system that enables real-time data analysis and
reduction for live cell analysis using fluorescence microscopy. We propose a novel system architecture capable of
analyzing a large amount of samples during the experiment and thus greatly minimizing the post-analysis phase
that is the common practice today. By utilizing data reduction algorithms, relevant information of the target
cells is extracted from the online collected data stream, and then used to adjust the experiment parameters in
real-time, allowing the system to dynamically react on changing sample properties and to control the microscope
setup accordingly. The proposed system consists of an integrated DSP-FPGA hybrid solution to ensure the
required real-time constraints, to execute efficiently the underlying computer vision algorithms and to close
the perception-action loop. We demonstrate our approach by addressing the selective imaging of cells with a
particular combination of markers. With this novel closed-loop system the amount of superfluous collected data
is minimized, while at the same time the information entropy increases.
We have developed a novel platform-concept, which allows the construction of an automated upright light-microscope-based
slide-scanning device. In order to achieve maximal speed at maximal image quality we have paid special attention
to a highly rigid, highly stiff construction. By using novel materials we have been able to achieve vibration-damping
characteristics many times better than conventional approaches. We have combined these new concepts with a voice-coil
based digitally controlled focus drive, which achieves the speed and stability of a piezo-element, yet allows travels of up
to 10 mm. Using scan-modes which avoid stop & go but instead keep the slide moving at a constant speed we have been
able to cut scan-times to levels approaching that of multi-objective approaches, yet with much higher flexibility and
better image quality.
The goal was to develop a light-microscope platform concept, which allows characterization of live cells in microtiter
plates or in live-cell slide chambers with a speed, sensitivity and versatility unattainable so far.
The goal was achieved by combining several novel technological concepts: a model-based digital control for a voice coil
focus drive; scanner technology to follow a continuously moving sample during image acquisition, thus avoiding the
usual stop-and-go; fast sectioning capabilities by using slit-scan confocal concepts; motorized dual emission image
registration; and integrated environmental control.
The goal was to develop an upright microscope platform for the screening of slides employing one- and two-photon laser
scanning techniques. A highly compact, vibration damping unit was created, which combines novel concepts for moving
a slide in three dimensions, keeping it focused while doing so, scanning a laser-focus over the sample using novel
galvanometer-control concepts, combining and separating excitation and emission beam and spectrally dispersing the
emitted light by a linearized prism-spectrograph. Spectral detection is achieved by turning a 128 x 128 back-thinned EMCCD
detector in a continuously reading spectral point-detector. To make the unit even ore versatile, it can be turned into
a conventional wide field fluorescence microscope, enabling rapid routine observation to select regions of the sample for
a subsequent, more detailed confocal analysis.
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