Multiplexing Phenotype and Function for More Biologically Relevant Insights - 27

Related Article from
"But when screening millions of compounds, one cannot be too complex," cautions
Dr. Stephan. "Screening has to be done fast because drug development is a race."
In a recent study that discussed the sustainability of current drug discovery
approaches, Dr. Stephan and colleagues highlighted the key benefits and
challenges of high-content analysis for phenotypic drug discovery. The study
considered how high-content capabilities, such as image capture and processing
and data analysis, could be incorporated into large-scale discovery efforts while
sustaining sufficient throughput.
"We need a model that in some way recapitulates what is going on in the body,"
insists Dr. Stephan. "But that is very difficult, and even more so in high-throughput
or high-content screening."
For example, for decades, cells have been grown using two-dimensional culture
systems, which do not reproduce the physiology and the elasticity of human tissue.
Three-dimensional culture systems model the physiology of live tissues more
accurately, but their use is still at relatively early stages.
"To reproduce physiological conditions, one has to mix different types of cells," says
Dr. Stephan. "The main limitation is how to reproduce the complexity of the human
body in a well."
Even when an ideal experimental model is in place, developers must deal with
another difficulty: selecting the best readout. Examples of readouts include the
size of the nucleus, the intensity of a specific stain, the presence of an antibody in a
specific cellular compartment, or the movement of the cell.
One of the strengths of high-content screening is the possibility of measuring
multiple parameters at the same time. However, virtually all readout types face
technical limitations. For example, the number of wavelengths that can be resolved
by a microscope is normally limited to four. The limited number of choices for some
readouts can bias the biological question that is being explored, and an in-depth
knowledge of the biology is therefore critical to allow the selection of the best
readout.

27

| January, 2019

In high-throughput screening positive
controls are needed at multiple steps, such
as assessment of plate quality or optimization of
experimental design.


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Multiplexing Phenotype and Function for More Biologically Relevant Insights

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