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Creating Higher Content

By R&D Editors | January 25, 2012

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PerkinElmer’s HCA ImagAmp reagent kits improve the throughput of screening and can be coupled with the company’s Opera and Operetta imaging systems.  

Whenever possible, drug researchers want one process to quickly reveal a collection of information. This makes high-content screening (HCS) and analysis (HCA) increasingly important in drug discovery. These techniques enable the automated collection of multi-parametric data, which deliver a more complete picture of the biology than traditional assays. Researchers can obtain all of these capabilities with the HCA ImagAmp reagent kits from PerkinElmer (Waltham, Mass.).

In short, HCA combines automation with microscopy. Cells flow by a microscope, which captures images that can be automatically analyzed. According to Aaron Risinger, imaging sales specialist at PerkinElmer, “High-content screening provides an unbiased approach to image segmentation and analysis.”

When using HCA ImagAmp reagent kits, says Pat Mayer, portfolio director, reagents research at PerkinElmer, the biggest benefit is sensitivity. “It let’s you detect signals where conventional technology has not been successful,” she says. Depending on the assay, the sensitivity can increase by 10- to 100-fold, according to Mayer. In addition to the improvements in sensitivity, it takes less time to capture an image with these kits. “That lets researchers perform more reactions, get more data in less time, and speed up the drug-discovery process,” says Mayer. To quantify the increase in speed, she says that the sensitivity of these kits can reduce the imaging exposure time by 10-fold, which increases the throughout by the same amount.

Researchers can also reduce the consumption of primary antibodies with these kits. It is very common, says Mayer, to reduce primary antibody concentration by 20-fold when adding HCA ImagAmp to an existing assay, with simultaneous improvement in sensitivity and shorter exposure times.

Obtaining these advantages, though, doesn’t take much more time or trouble. These kits work like other HCA processes, except that they require one added step: a 10-minute incubation. In fact, researchers can also use the same fluorophores that they’ve been using in other approaches to HCA. As Mayer says, “It’s easy to integrate these kits into the current workflow.”

These kits work in PerkinElmer platforms, including the Opera and Operetta imaging systems, but they also work with high-content platforms from other companies. Furthermore, the HCA ImagAmp kits consist of a family of three different fluorophores. As Mayer says, “You can use more than one at a time, so they can be used in multiplexed assays.

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These U2OS cancer cells—part of the Thermo Scientific ToxInsight Endocrine Profiler Panel—were bioengineered to express a GFP-tagged androgen receptor. Then, the cells were left untreated (left) or stimulated for endocrine activity (right) and imaged on the Thermo Scientific ToxInsight IVT Platform.  

Screening for endocrine-active chemical
The safety testing done in the pharmaceutical industry involves more than just compounds. Even the plastics used to make the bag for intravenous drugs must be tested. Those materials can contain endocrine-active compounds, according to Mark Collins, PhD, director of global marketing for the Cellomics business unit of Waltham, Mass.-based Thermo Fisher Scientific. To simplify testing materials for such compounds, Thermo Fisher developed the Thermo Scientific ToxInsight endocrine profiler panel.

In describing this product, Collins says, “It consists of assays that look at the effect of endocrine-active chemicals on cells.” He adds, “We put together these panels using bioengineered cell lines and fluorescent dyes.” The panels determine whether a chemical interacts with androgen or estrogen receptors. “This is high-throughput technology for endocrine-active chemicals, that allows more to be screened while reducing the use of animals in traditional tests,” Collins says.

A researcher could use these panels to screen for chemicals that might make good drugs because they stimulate endocrine activity. On the other hand, a researcher might screen a medical device to make sure that it doesn’t affect endocrine systems.

Simplifying the workflow for high-content analysis helps drug researchers apply this technology in more ways, which reveal compound features faster.

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