3D Deconvolution

Out-of-focus intensities are present in all acquired images. These intensities can be accounted for by observing the behavior of light originating in a point source and passing through the microscope optics. This behavior is described by the Point Spread Function (PSF). It can be used to quantitatively compensate for the blurring of images due to out-of-focus information. This process is called deconvolution. MetaMorph's 3D Deconvolution module from AutoQuant helps improve images by reassigning out-of-focus intensities back to the spatial locations to where they originated. The results are images with sharper definition and lower background, better contrast, and improved signal-to-noise ratio.

3D Reconstruction

The 4D Viewer enables the visualization of multi dimensional data sets, time lapse or Z stacks. Users can simultaneously view multiple Z sections as a 3D reconstruction, with multiple , wavelengths, time points, and positions in a single intuitive viewing window. Users can interactively rotate the 3D view and obtain volumetric measurements. A 3D model consisting of rotated views from a stack of images can also be created. Using a stack of planes from a Z-series, users may configure the angle, orientation, Z-axis distance, and reconstruction type for the model.

Cell Migration/Cell Proliferation

The Count Nuclei Application Module is designed to automate accurate counting of nuclei for most types of cells. The module counts nuclei even when the background is uneven, providing superior segmentation compared to simple thresholding. This module can be used to count nuclei across large data sets and log the results to a spreadsheet.

Deconvolution

For 2D deconvolution, MetaMorph offers No Neighbors and Nearest Neighbors algorithms to remove out-of-focus information from individual planes or a Z-stack. Both modules use an estimated three-dimensional Point Spread Function (PSF). The 3D deconvolution module can be used for analysis of Z-stacks and may be configured to use either a Blind algorithm which estimates a Point Spread Function (PSF) or an algorithm where a measured PSF which characterizes the objective lens is provided.

Densitometry

MetaMorph can perform quantitative densitometry. The software will display the optical densities of a brightfield source image in a scaled 8-bit or 16-bit image.

Fluorescence Overlay

To better visualize events, up to six fluorescence images can be overlaid over a background Differential Interference Contrast (DIC) or phase contrast image.

Fluorescent in situ Hybridization (FISH)

FISH is used for the detection of target DNA or RNA molecules with a system of coupled fluorochromes. The detection of nucleotidic sequences on a combed DNA molecule is performed indirectly, by first hybridizing the sought nucleotidic sequences with the combed DNA (also called the matrix DNA or target). If the probes are synthesized with incorporated fluorescent molecules or antigenic sites which can be recognized with fluorescent antibodies, the direct visualization of the relative position of the probes is possible. MetaMorph easily automates the process of acquiring, color combining, and visualizing multiple wavelengths from FISH and immunocytochemistry experiments.

Fluorescence Immunocytochemistry

Immunocytochemistry is the in situ detection and demonstration of cellular constituents using specific antigen-antibody reactions. It has evolved to become an integral aid in modern histopathology. Crucial to its success has been the development of technologies that have allowed the highly sensitive and reliable detection of cellular markers within routinely fixed and processed samples. MetaMorph for Olympus can acquire, combine, and measure images of one or more wavelengths from immunocytochemistry and in situ hybridization experiments. The morphological measurement tools in MetaMorph are ideal for processing immunohistological tissue samples. MetaMorph for Olympus can acquire, combine, and measure images of one or more wavelengths from immunocytochemistry and in situ hybridization experiments.