Label-free quantitation of peptide release from neurons in a microfluidic device with mass spectrometry imaging.

Citation data:

Lab on a chip, ISSN: 1473-0189, Vol: 12, Issue: 11, Page: 2037-45

Publication Year:
2012
Usage 6
Abstract Views 6
Captures 28
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Citations 29
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Repository URL:
http://scholarworks.unist.ac.kr/handle/201301/6950
PMID:
22508372
DOI:
10.1039/c2lc21085a
PMCID:
PMC3558029
Author(s):
Zhong, Ming, Lee, Chang Young, Croushore, Callie A., Sweedler, Jonathan V.
Publisher(s):
Royal Society of Chemistry (RSC), The Royal Society of Chemistry, ROYAL SOC CHEMISTRY
Tags:
Biochemistry, Genetics and Molecular Biology, Chemistry, Chemical Engineering, Engineering, MALDI-TOF MS, LASER-DESORPTION/IONIZATION-TIME, BAG CELL NEURONS, CAPILLARY-ELECTROPHORESIS, NEUROENDOCRINE CELLS, GLYCOL) MONOLAYERS, HORMONE-SECRETION, HIGH-THROUGHPUT, SOLID-SURFACES, SINGLE-CELL
article description
Microfluidic technology allows the manipulation of mass-limited samples and when used with cultured cells, enables control of the extracellular microenvironment, making it well suited for studying neurons and their response to environmental perturbations. While matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) provides for off-line coupling to microfluidic devices for characterizing small-volume extracellular releasates, performing quantitative studies with MALDI is challenging. Here we describe a label-free absolute quantitation approach for microfluidic devices. We optimize device fabrication to prevent analyte losses before measurement and then incorporate a substrate that collects the analytes as they flow through a collection channel. Following collection, the channel is interrogated using MS imaging. Rather than quantifying the sample present via MS peak height, the length of the channel containing appreciable analyte signal is used as a measure of analyte amount. A linear relationship between peptide amount and band length is suggested by modeling the adsorption process and this relationship is validated using two neuropeptides, acidic peptide (AP) and α-bag cell peptide [1-9] (αBCP). The variance of length measurement, defined as the ratio of standard error to mean value, is as low as 3% between devices. The limit of detection (LOD) of our system is 600 fmol for AP and 400 fmol for αBCP. Using appropriate calibrations, we determined that an individual Aplysia bag cell neuron secretes 0.15 ± 0.03 pmol of AP and 0.13 ± 0.06 pmol of αBCP after being stimulated with elevated KCl. This quantitation approach is robust, does not require labeling, and is well suited for miniaturized off-line characterization from microfluidic devices.