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Application modules are intended to simplify complex, but routine, processes:
The Extended Focal Imaging (EFI) module allows the user to create a single in-focus image from several images with different focal planes. By employing contrast algorithms, the software uses the sharpest pixels to create the final image. This is particularly useful for thick or unevenly mounted tissue specimens and whole or partial insects. The optional height map feature lends a three-dimensional view to the final image. EFI is available in the Basic Edition and higher.
Example of EFI
Intensity-based height map (filtered)
Extended Focal Imaging (EFI)
As one of our most popular modules, the Multiple Image Alignment (MIA) module allows the user to acquire and assemble a high-resolution image from a series of adjacent images using pattern matching algorithms. MicroSuite™ FIVE offers a new option: the ability to collect multiple Z-planes at each XY location, through a simple check box that enables the Extended Focal Imaging (EFI) module within the MIA module. This results in a stitched image that is completely in focus, regardless of variability in specimen thickness. MIA is available in the Basic Edition and higher.
For systems equipped with a motorized XY or XYZ stage, the interactive Stage Navigator can be used to collect overview images of entire slides, with or without overlap between the images. The module allows users to load individual images or areas of interest at full resolution, record positions and add comments, or point-and-click on the overview image to return to a particular XY location and view the area at a higher magnification. Areas of interest within the overview image can also be rescanned at a higher magnification. The Stage Navigator module is available in the Analytical, Biological, and Advanced Suites.
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The Stage Manager/Automater module can also be used with a motorized stage, to create and save stage positions for re-visitation and automated image acquisition and analysis. The user can define processing routines, such as particle analysis, and carry out these processes in an automated fashion. Results can be displayed in spreadsheets, tables, and histograms, and can easily be incorporated into reports using the Report Generator function of the software. The Stage Manager/Automater module is available in the Advanced Suite, but can be added on to other packages.
The Multiple Fluorescence Information Processing (MFIP) module is designed to facilitate the acquisition and processing of multi-colored fluorescence images. Applications include the labeling of cells and organelles with multiple fluorescent dyes, Fluorescence in situ Hybridization (FISH), and gene expression studies with fluorescent proteins. MFIP also supports the overlay of a transparent grayscale image (DIC, phase contrast, brightfield, or darkfield) with fluorescence images.
Example of MFIP
New visualization tools permit the user to adjust the brightness, contrast, and gamma of single color or composite images, allowing for the optimization of each channel relative to the others. The Auto Adjust Display button adjusts the display for all images in a stack, relative to the best image in the stack. The MFIP module is available in the Basic Edition and the Biological, and Advanced Suites.
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Spectral Unmixing is a unique tool that allows the separation of spectrally similar fluorochromes such as CFP and YFP. The software first calculates a ratio from reference images of single-labeled specimens captured with the same filters and imaging conditions as the dual-labeled specimens. After a background subtraction, the unmixing algorithm applies this ratio to images from the dual-labeled specimens. The Spectral Unmixing module is available in the Biological and Advanced Suites.
Under license from AutoQuant, the Rapid Image Deconvolution (RIDE) module includes three types of filters for the deconvolution of light microscope images: No Neighbor, Nearest Neighbor, and Inverse Filter. RIDE is included with the Advanced Suite. The 3D Widefield Deconvolution and 3D Confocal Deconvolution add-in modules incorporate AutoQuant's constrained iterative 3D blind deconvolution algorithms for widefield and confocal image stacks respectively. Image stacks can be imported or automatically collected on a microscope with a motorized Z drive. All Olympus FluoView image stacks can be opened with MicroSuite™ FIVE. Various operations on deconvolved image stacks are possible, such as applying pseudocolors, smoothing the images, and selecting regions of interest. Visualization tools include the Slice Viewer, for projecting stacks in the X, Y, and Z directions, and the Voxel Viewer, for stereo rendering, iso-surface projection and 3D animation.
The WellNavigator add-in module integrates a motorized microscope, stage, and camera to automatically acquire, process and archive images from various sized well plates, microtiter plates, or chamber slides. Acquisition options include autofocus, shading correction, EFI (Z-stack), and mFIP (multi-channel fluorescence). All the standard tools for particle analysis are also available. The Experiment Manager allows for easy customization of routine analytical procedures. Calibration of new well plates or slides can be done automatically or with user assistance.
The Image Sequence Processing add-in module for multichannel (XYZT, color) acquisition controls and coordinates the actions of Olympus motorized microscopes and third-party motorized stages, filter wheels, and shutters. The user can interactively define X, Y, and Z positions, autofocus parameters (select cameras), microscope parameters, and time parameters. Acquired sequences can be visualized in the embedded animation studio.
TrackIt! is a new add-in module that tracks multiple objects in 2D space. Trajectories, speed, and distances can be measured with this module.
As one of our most popular modules, the Multiple Image Alignment (MIA) module allows the user to acquire and assemble a high-resolution image from a series of adjacent images using pattern matching algorithms. MicroSuite™ FIVE offers a new option: the ability to collect multiple Z-planes at each XY location, through a simple check box that enables the Extended Focal Imaging (EFI) module within the MIA module. This results in a stitched image that is completely in focus, regardless of variability in specimen thickness. MIA is available in the Basic Edition and higher.
