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Assays for high content imaging

Here is a non-exhaustive list of the HCI assays that can be run in our laboratory.

Angiogenesis

Angiogenesis, the formation of blood vessels, underlies many normal and pathological processes, including responses to toxicity. This assay identifies angiogenic tubes (micro-capillaries) formed by endothelial cells, and provides quantitative measurements related to angiogenic tube formation such as the number of tubes, their morphology and branching, and an Angiogenic Index.

Apoptosis

Apoptosis is the process of programmed cell death in which a tightly controlled cascade of events occurs in order to maintain tissue homeostasis. This assay measures key events in the apoptotic cascade and can also be used to distinguish apoptosis from necrosis.

Cell Cycle

The Cell Cycle Assay allows determination of the stage of the cell cycle in a sample of cells. A cell replicates by performing an orderly sequence of events in which it duplicates its genetic contents and then divides into two; this cycle of duplication and division is known as the cell cycle.

Cell Health Profiling

Measurement and characterization of cell health is critical for assessment of toxicity. This assay enables examination of a variety of cell health markers including cell viability, cytotoxicity, and apoptosis by evaluating morphological and intensity changes within cellular compartments.

Cell Toxicity

Following a toxic insult, cells may undergo either necrosis or apoptosis. This assay evaluates both processes by quantifying changes in nuclear size, morphology, cell permeability, as well as functional changes to cellular organelles.

Cell Viability

Live and dead cells represent two physiological states that are well suited to quantitative analysis with this BioApplication.  This assay quantifies (percent of) dead cells, live cells, or both cell populations, as well as general intensity outputs are given

Cholestasis

Cholestasis is the result of impairment in bile flow that leads to abnormal retention of components of bile in the liver and serum. The cholestasis assay utilizes an in vitro method of canal formation and the ability of cells to metabolize and export a fluorescent substrate that accumulates in the canaliculi. Readout measurements for the assay can include intensity, number and complexity of the canalicular network.

Colony formation

Some cells grow in tight groups or colonies rather than individually, as the need for close proximity can affect the cell's growth, appearance, protein expression, and function.  This assay identifies and analyzes colonies, and reports their size, morphology, the number of cells in the colony as well as in the colony's interior or periphery, and the presence of other markers.

Cytoplasm to Membrane Translocation

Proteins in cells often follow a specific localization either as a prelude to, or result of, being acted upon. Signal transducers are members of this group and can, for specific cases, relocate from the cytoplasm of the cell to the cell membrane as a response. This assay evaluates the localization of intensity in both compartments.

DNA Damage

DNA damage causes chromosomal instability that leads to oncogenesis, apoptosis, and severe failure of cell functions.  This assay measures changes in intensity within nuclei or foci sites of treated cells to help determine if DNA damage has occurred and to what extent.

DNA Damage (Comet)

Exposing cells to DNA damaging agents can result in the formation of a nucleic acid comet following electrophoresis in low-melting agarose. This assay quantifies the size and shape of the comets to specifically assess the extent of DNA breakage or cross-linking caused by compound treatment or other genotoxic assault.

Drug Induced Liver Injury

The Drug Induced Liver Injury (DILI) Assay monitors hepatotoxicity by simultaneous detection and analysis of multiplexed cellular targets and properties associated with cell loss, cellular redox stress, and mitochondrial stress.  Differences in fluorescent intensities and location of the biomarkers in liver cell models are used to assess the hepatotoxic effects of compound treatment.

General Colocalization

The General Colocalization assay/application has the ability to measure overlap of targets/markers in any of the four independent imaging channels of the HCI platforms. The application measures and reports out features that are relevant to overlap based upon intensity or area of the regions of interest that are relevant to the biological studied.