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High-Content Analysis

High-content analysis (HCA) or high-content screening (HCS) combines the molecular tools of cell biology with automated high-resolution microscopy, robotic handling, and automated analysis.

HCA provides multiplexed, quantitative data based on automated cell imaging and analysis. Complex questions are answered quickly in a true biological context, giving deeper insights into biological systems.

High-content analysis  involves:

  1. Probing cells in microtiter plates or on slides with single or multiple fluorescent sensors.
  2. Using a high-resolution imaging instrument for rapid capture of cell images.
  3. Extracting detailed information from the images with powerful analysis software.
Visualize Your Research Deeper Insights, Fast Advantages of High-Content Analysis
Human neural stem cells from fetal cortex

High-content analysis enables detailed study of cellular mechanisms and interactions in multiple samples. Examples include studying whether cell division, motility, or protein uptake is affected when cells are exposed to external stimuli such as chemical inhibitors.

In basic research, information generated using HCA is highly valuable to gain a deeper understanding of complex diseases, including cancer, neurology, immunology, and infectious diseases. In drug discovery research, the throughput gained using HCA makes it a valuable technology for identifying targets, for example through the use of RNAi screens (HCS). Deeper insights provided by HCA give more information on the mode of action or toxicity profiling, enabling better decisions to be made on which candidates to progress with.

Detailed examination of cells at the microscopic level enables probing complex signaling mechanisms, elucidating molecular function, studying cell behavior, and correlating multiple events and markers against the same physiological background.

With HCA:

  • Increase information content and throughput by using multiple probes in one assay.
  • Reduce assay times by days to hours or hours to minutes.
  • Obtain more relevant data by investigating live intact cells (rather than cell extracts).
  • Ask complex questions regarding interdependent biological events/processes that would be impossible with traditional technologies.

Alternatives to HCA include biochemical assays, conventional microscopy, Western blotting, and flow cytometry. While these techniques can be highly valuable, they can also have limitations in throughput, can disrupt cell integrity, and be labor intensive. These limitations reduce the number and complexity of assays, and ultimately reduce the relevance of the results in a biological context.

HCA allows you to capture and analyze tens of thousands of images in a day, where each image contains many hundreds of cells, monitoring a hundred or so parameters. This makes it possible to leverage the power of microscopy to perform large-scale screens and functional studies rich in multiparameter subcellular information. Application of HCA for screening large libraries of compounds or genetic material is referred to as high-content screening (HCS).

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