Developed in conjunction with researchers, MetaMorph Software offers a range of tools for a variety of specific applications. These MetaMorph Software Applications Modules are tailored to make it easy to perform the common, but sometimes complex, specialized analysis.
- Micronuclei Application Module
-
Micronuclei Application Module classifies cells based on nuclear morphology for genotoxicity applications but is also ideal for cell health or multi-nucleated cell populations with additional markers for analysis of apoptosis, necrosis or to distinguish any small structure next to a large one such as a yeast bud.
- Highly accurate classification of cells (micro-, mono-, bi-, or multi-nucleate) achieved with proprietary algorithm to discriminate phenotypes based on cell morphology, number of nuclei, distance of micronuclei from nucleus, micronuclei vs. “blebs” or “buds”.
- Only requires a single fluorophore (nuclear stain) to identify cells in various classifications, eliminating the requirement for a cytoplasm stain, thereby reducing sample preparation, image acquisition and analysis time.
- Use of two additional fluorophores facilitates further phenotypic analysis, such as transfection markers to identify transfected cells or kinetochore markers to differentiate micronuclei originating from clastogens (which create acentric chromosomal fragments) or from aneugens (which cause whole chromosomal losses).
Images of cell nuclei before (left) and after (right) image segmentation.
Micronuclei, binucleate, and mutinucleate cells detected by the application module are highlighted.
In total, more than 60 measurements per image and 30 measurements per cell are available to assist with identification of the wide range of phenotypes associated with genotoxicity studies.
- Neurite outgrowth Application Module
-
Neurite Outgrowth Application Module is designed to measure and characterize outgrowths, the extension of processes from the cell body, which are natural parts of neuronal development. Inhibition or stimulation of neurite outgrowth is implicated in a broad range of neurological disorders or injuries including stroke, Parkinson’s disease, Alzheimer’s disease, and spinal cord injuries.
- Unique assay that cannot be performed without imaging
- Provides consistent results faster than manual tracing and counting
- Output parameters, on a per field-of-view or per cell basis may include outgrowth count, mean or maximum lengths, branches, straightness, cell number or cell body size, or % with significant growth
(Left) Images of fluorescently stained neurons. (Right) After image segmentation.
Each outgrowth is assigned to a cell body. All the outgrowths and cell bodies are then measured.
(Courtesy of Kris Poulsen and Davide Foletti, Rinat Neuroscience Corporation).
- Angiogenesis Tube Formation Application Module
-
Angiogenesis Tube Formation Application Module facilitates the analysis of tube formation, a model experimental system for angiogenesis. The promotion or inhibition of blood vessel formation is one focus for cancer, diabetes and other vascular disease research.
- Captures the three-dimensional behavior of tubes utilizing Z-stack acquisition available through the MetaXpress Software
- A best-focus image of the z-stack is analyzed to characterize tubes by area, length, branch points, and nodes
Human Mammary Epithelial Cells (HMEC-1) tube formation.
(Left) 3D Acquisition (Center) Best focus algorithm collapses z-series into a single image (Right) tubes and nodes are identified with the application module (Data courtesy of BD Biosciences)
- Mitotic Application Module
-
Mitotic Index Application Module is designed for the quantitative discrimination of mitotic and interphase cells, a critical tool for oncology drug discovery programs, providing insight into potential anti-cancer therapeutics that rely on arresting mitosis in cancerous cells to prevent uncontrolled proliferation.
(Left) CHO-K1 cells treated with Nocodazole for 18 hours before staining with a marker for mitotic arrest.
(Right) After image segmentation the green mask identifies mitotic cells and red indicates nuclei in interphase.
- Cell Cycle Application Module
-
Cell Cycle Application Module classifies and quantifies cells in various stages of the cell cycle to investigate cell cycle progression. In healthy non-cancerous cells, challenges with DNA damage, hypoxia, metabolic changes or spindle disruption result in the triggering of checkpoints and cell cycle arrest. Cancerous cells commonly lose checkpoints and divide uncontrollably, even in challenging conditions. With the appropriate tools, researchers can screen for drugs that cause cell cycle arrest or cell death in cancerous cells.
- Differentiates 5 phases of the cell cycle using only a nuclear stain (G0/G1, S, G2, Early or Late M).
- Optional detection of mitosis with specific fluorophores to better distinguish M-phase cells when using low magnification.
- Optional detection of apoptotic markers to detect conditions that trigger apoptosis.
- Interactive color-coded graphs to easily set classification cutoffs.
Classification and quantification of cells in 5 phases of the cell cycle.
Interactive color coded graphs allow to easily set classification cutoffs.
- Monopole Detection Application Module
-
Monopole Detection Application Module monitors the disruption of the formation of bipolar spindles, a successful mechanism used by drugs such monastol to stop the progression of cancerous cells through mitosis. The module quantifies mitotic cells with monopolar or bipolar spindles in cells stained with a DNA probe and a microtubule probe.
(Left) 3T3-L1 mouse fibroblast cells treated with monastrol and stained for beta tubulin.
Nuclei are stained with Hoechst. (Right) the module identifies interphase cells (red), bipolar spindles (blue) and monopoles (green).
- Cell Scoring Application Module
-
Cell Scoring Application Module is a general and flexible solution to identify subpopulations of cells tagged with a second fluorescent probe and is ideal for examining transfection efficiencies, pathway activation (kinases) or adipogenesis.
- Label all cells with a nuclear or whole cell stain and only cells-of-interest will exhibit staining of the target with a second fluorophore
- Robust identification of cells exhibiting markers for DNA damage, differentiation, or other selective activation
- Equally accurate at low or high magnification
- Output results as number or percent positive or negative or include fluorescence intensity values of each cell or as a total or average of the field-of-view.
(Left-top) HeLa cells labeled with DAPI.
(Left-bottom) Immunostaining for target marker.
(Center-top) Cell Scoring identifies all nuclei (red).
(Center-bottom) Cell Scoring Application Module identifies immunostaining positive.
(Right) Overlay shows cells positive for marker (green) and negative for marker (red).
- Count Nuclei Application Module
-
Count Nuclei Application Module automates accurate counting of nuclei for most types of cells and are ideal for the study of cell proliferation, cell counting or cell migration. These modules count nuclei even when the background is uneven, providing superior segmentation compared to simple thresholding.
- Module identifies nuclei, taking into consideration nuclear size and varying background intensities
- Touching objects are automatically split
- More consistent between users and faster than manual counting.
- Easier and faster than flow cytometry, which is a low-throughput method that requires trypsinization of cells
(Left) Image has uneven illumination and touching cells. (Right) the module identifies touching cells as separate objects (as indicated by the red arrow).
Adaptive Background Correction compensates for the variable background intensity – even when background intensity in one area of the image is greater than nuclei intensity in another area.
- Cell Health Application Module
-
Cell Health Application Module classifies cells states in viable, early apoptosis, late apoptosis or necrosis.
- 3 wavelength detection of nuclear or cytoplasmic staining
- Scores cells as viable, necrotic, early/late apoptotic based on intensity of markers
- Site-by-site data including counts, percentages, total and average area and intensity
- Cell-by-cell data including health status, area and intensity measurements
- Validated with most common cell health dyes such as Yo-Pro 1, Annexin V or CellEvent Caspase 3/7 for apoptosis, Propidium Iodide for necrosis, or mitotoxicity dyes such as JC-1 or JC-10 to study loss of mitochondrial potential
(Left) Cells were treated with 1 μM Staurosporine for 12 hours and stained with Hoechst 33342, YO-PRO-1 and PI (Right) Image analysis results are shown as a colored overlay on the source image.
Viable nuclei are shown in green.
Early apoptotic nuclei are shown in blue.
Late apoptotic nuclei are shown in purple.
Necrotic nuclei are shown in red.
- Live/Dead Application Module
-
Live Dead Application Module is compatible with commercially available Live/Dead assay kits designed to study cell proliferation or death. Common applications monitor cell proliferation associated with cancer or cell death due to neuromuscular diseases such as Alzheimer’s and Parkinson’s Disease or in response to cytotoxic and apoptotic events.
- 2 wavelength present in any part of the cell, not necessarily the nucleus
- Scores cells as live or dead based on intensity of markers
- Site-by-site data including counts, percentages, total and average area and intensity
- Cell-by-cell data including classification, area and intensity measurements
(Left) Cells were treated with 1 µM Staurosporine.
(Right) Red objects are live cells and green objects are dead.
- Granularity Application Module
-
Granularity Application Module facilitates the analysis of punctate structures such as the one observed during clustering of target molecules for receptor internalization or within the nucleus or even the punctate patterns of mitochondria.
U2OS cells (left), overlay (right).
The module identifies granules and nuclei, even in cells with high background (red arrow).