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I
am studying the role of soluble signaling factors
in cancer development using the mouse breast
cancer model. The study of soluble factors is
complicated by several factors. First, many
soluble factors are expressed at low levels
and these factors are often dissipated in large
culture vessels. We culture the cells within
microfluidic channels in order to minimize the
dilution of signaling factors that are expressed
at low levels, providing an environment that
more closely resembles in-vivo conditions.
The
complex nature of cell culture media also posses
a challenge. Typical media contains an assortment
of proteins, growth factors, amino acids, lipids,
carbohydrates, hormones, vitamins, and many
other chemicals. Further, the concentration
of these compounds ranges from ng/ml to mg/ml;
low concentration species are difficult to measure
due to both the limits of detection and masking
by more abundant species such as albumin. I
am currently evaluating sample preparation methods
that can be used in conjunction with sensitive
detection methods, such as mass spectrometry
and fluorescence, so that we can study signaling
factors that are expressed at low levels.
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