Modern high-content screening microscopes enable fast, automated imaging of whole microtiter plates by imaging fixed positions inside different wells. This is perfect for in-vitro cell culture-based readouts and other tests with evenly distributed phenotypes. However, it becomes limited when large specimens or rare events are examined, as the limited field of view (FOV) of high-magnification objectives might not fully encompass the area of interest.
Researchers are frequently limited to lesser magnification acquisition, resulting in data with low resolution or the omission of features of interest in multiple wells. This article explains how Bruker's Acquifer Imaging Machine (IM) overcomes these issues with PlateViewer software and provides a semi-automated technique for experiments involving advanced supervised feedback microscopy.
Automated Experimental Approaches
For tissue-specific imaging in large specimens (e.g., zebrafish), a method that can automatically identify and zoom in on the tissue or organ of interest is required. Fully automated tissue detection and imaging frequently require complex image processing routines to be developed. Each project requires careful balancing between software development time and project size.
Semi-automated methods represent an optimal solution, requiring minimal user interaction and no custom algorithm development. This simpler alternative can easily handle even complicated or variable structures that would typically demand extensive development of image detection routines. This method is useful for screening projects as they start instantly, saving both time and resources.
Plate-Viewer Software
Figure 1. Illustration of Click-Tool functionality. (a) Zebrafish embryo imaged with a 2x objective and visualized in Plate-Viewer. The crosshair is centered on the heart region in a three-day-old embryo of the epi:GFP;myl7mR transgenic line. The red bounding box indicates the FOV of a 10x objective used for subsequent high-resolution imaging. (b) Single Z-plane of a high-resolution dataset is automatically acquired on Acquifer IM. Image Credit: Bruker Nano Surfaces and Metrology
Plate-Viewer software provides a semi-automated method for supervised feedback microscopy. Low-magnification pre-screen data of a full microtiter plate is visualized after the plate layout for a rapid and easy-to-understand overview. The integrated “click-tool” function permits assay experts to select areas of interest (ROIs) for different wells.
Furthermore, in-built and generic “template-matching” algorithms facilitate robust automatic localization of several target structures. These include complicated reporter expression patterns, morphological features, and rare events in different wells. Acquifer IM spontaneously gathers information at a high resolution from specific areas that correspond to predefined settings, and acquired high-resolution data is then easily and intuitively visualized in Plate-Viewer.
Plate-Viewer has multiple distinct properties that make it possible for users to:
- Select ROIs to be automatically imaged by the click of a mouse or template matching
- Preview FOVs of higher magnification objectives via an adjustable bounding box
- Intuitively visualize and search through large-scale screening datasets
- Easily search through sophisticated multidimensional datasets
- Apply lookup tables and overlay channels to enhance manual inspection
- Change basic image parameters to improve the visualization of details
- Plug-in interface for integration of external image processing instruments
Figure 2. Overview of the Plate-Viewer functionalities. Visualization of identical planes of all available wells within a microtiter plate (left). Preview of the currently selected well coordinate (right). Control panels in the top-right corner enable the generation of colored overlay images and the display of selected channels. Control panels in the lower left corner allow channel-specific adjustment of the histogram and the choice of navigation mode through multidimensional datasets (e.g., slice, loops, or subpositions). Image Credit: Bruker Nano Surfaces and Metrology
Intuitive Solution for High-Content Screening
Acquifer IM is a complete solution for experiments involving supervised feedback microscopy. Its Plate-Viewer software makes use of a semi-automated method to simplify complicated high-content screening experiments. This dynamic tool allows researchers to more easily and reliably carry out tissue-specific imaging of complex and variable structures in large specimens.
About Bruker Nano Surfaces and Metrology
Bruker Nano Surfaces and Metrology provides high-performance, specialized analysis, and testing technology for the widest range of research and production applications.
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Whatever your measurement and analysis needs, whatever your material or scale of investigation, Bruker has a specialized high-performance solution for you.
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