Enzyme-linked
immunosorbent assay (ELISA) is the gold standard
method for protein biomarkers. However, scaling up ELISA for multiplexed
biomarker analysis is not a trivial task due to the lengthy procedures
for fluid manipulation and high reagent/sample consumption. Herein,
we present a highly scalable multiplexed ELISA that achieves a similar
level of performance to commercial single-target ELISA kits as well
as shorter assay time, less consumption, and simpler procedures. This
ELISA is enabled by a novel microscale fluid manipulation method,
composable microfluidic plates (cPlate), which are comprised of miniaturized
96-well plates and their corresponding channel plates. By assembling
and disassembling the plates, all of the fluid manipulations for 96
independent ELISA reactions can be achieved simultaneously without
any external fluid manipulation equipment. Simultaneous quantification
of four protein biomarkers in serum samples is demonstrated with the
cPlate system, achieving high sensitivity and specificity (∼
pg/mL), short assay time (∼1 h), low consumption (∼5
μL/well), high scalability, and ease of use. This platform is
further applied to probe the levels of three protein biomarkers related
to vascular dysfunction under pulmonary nanoparticle exposure in rat’s
plasma. Because of the low cost, portability, and instrument-free
nature of the cPlate system, it will have great potential for multiplexed
point-of-care testing in resource-limited regions.
Soluble stable radicals are used as spin probes and spin labels for in vitro and in vivo Electron Paramagnetic Resonance (EPR) spectroscopy and imaging applications. We report the synthesis and characterization of a perchlorinated triarylmethyl radical enriched 99% at the central carbon, 13 C 1 -PTMTC. The anisotropy of the hyperfine splitting with the 13 C 1 (A x =26, A y =25, A z =199.5 MHz) and the g (g x =2.0015, g y =2.0015, g z =2.0040) are responsible for a strong effect of the radical tumbling rate on the EPR spectrum. The rotational correlation time can be determine by spectral simulation or via the linewidth after calibration. As spin probe 13 C 1 -PTMTC can be used to measure media microviscosity with high sensitivity. Bound to a macromolecule as spin label, 13 C 1 -PTMTC could be used to study local mobility and molecular interactions. File list (2) download file view on ChemRxiv Manuscript_final.docx (1.07 MiB) download file view on ChemRxiv Supplementary_final.docx (452.34 KiB)
<p>We report the synthesis of
hydroxyethyl tetrathiatriarylmethyl
radicals OX063 and its deuterated analogue OX071 for biomedical EPR
applications.<br></p>
<p>We report the synthesis of
hydroxyethyl tetrathiatriarylmethyl
radicals OX063 and its deuterated analogue OX071 for biomedical EPR
applications.<br></p>
<p>Soluble stable radicals are used as spin probes and spin
labels for <i>in vitro</i> and <i>in vivo</i> Electron Paramagnetic Resonance
(EPR) spectroscopy and imaging applications. We report the synthesis and
characterization of a perchlorinated triarylmethyl radical enriched 99% at the
central carbon, <b><sup>13</sup>C<sub>1</sub>-PTMTC</b>.
The anisotropy of the hyperfine splitting with the <sup>13</sup>C<sub>1</sub>
(A<sub>x</sub>=26, A<sub>y</sub>=25, A<sub>z</sub>=199.5 MHz) and the g (g<sub>x</sub>=2.0015,
g<sub>y</sub>=2.0015, g<sub>z</sub>=2.0040) are responsible for a strong effect
of the radical tumbling rate on the EPR spectrum. The rotational correlation
time can be determine by spectral simulation or via the linewidth after
calibration. As spin probe <b><sup>13</sup>C<sub>1</sub>-PTMTC
</b>can be used to measure media microviscosity with high sensitivity. Bound to
a macromolecule as spin label, <b><sup>13</sup>C<sub>1</sub>-PTMTC
</b>could be used to study local mobility and molecular interactions.</p>
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