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Localized in situ hydrogel-mediated protein digestion and extraction technique for on-tissue analysis.
Harris GA, Nicklay JJ, Caprioli RM
(2013) Anal Chem 85: 2717-23
MeSH Terms: Animals, Cerebrum, Chemical Fractionation, Chromatography, Liquid, Hydrogels, Molecular Weight, Proteins, Proteolysis, Rats, Rats, Sprague-Dawley, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Tandem Mass Spectrometry
Show Abstract · Added March 7, 2014
A simultaneous on-tissue proteolytic digestion and extraction method is described for the in situ analysis of proteins from spatially distinct areas of a tissue section. The digestion occurs on-tissue within a hydrogel network, and peptides extracted from this gel are identified with liquid chromatography tandem MS (LC-MS/MS). The hydrogels are compatible with solubility agents (e.g., chaotropes and detergents) known to improve enzymatic digestion of proteins. Additionally, digestions and extractions are compatible with imaging mass spectrometry (IMS) experiments. As an example application, an initial IMS experiment was conducted to profile lipid species using a traveling wave ion mobility mass spectrometer. On-tissue MS/MS was also performed on the same tissue section to identify lipid ions that showed spatial differences. Subsequently, the section underwent an on-tissue hydrogel digestion to reveal 96 proteins that colocalized to the rat brain cerebellum. Hematoxylin and eosin (H & E) staining was then performed to provide additional histological information about the tissue structure. This technology provides a versatile workflow that can be used to correlate multiple complementary analytical approaches in the analysis of a single tissue section.
1 Communities
1 Members
0 Resources
12 MeSH Terms
A Unified attentional bottleneck in the human brain.
Tombu MN, Asplund CL, Dux PE, Godwin D, Martin JW, Marois R
(2011) Proc Natl Acad Sci U S A 108: 13426-31
MeSH Terms: Adult, Attention, Brain, Brain Mapping, Cerebrum, Decision Making, Female, Frontal Lobe, Humans, Magnetic Resonance Imaging, Mental Processes, Psychomotor Performance, Time Factors, Young Adult
Show Abstract · Added February 15, 2016
Human information processing is characterized by bottlenecks that constrain throughput. These bottlenecks limit both what we can perceive and what we can act on in multitask settings. Although perceptual and response limitations are often attributed to independent information processing bottlenecks, it has recently been suggested that a common attentional limitation may be responsible for both. To date, however, evidence supporting the existence of such a "unified" bottleneck has been mixed. Here, we tested the unified bottleneck hypothesis using time-resolved fMRI. Experiment 1 isolated brain regions involved in the response selection bottleneck that limits speeded dual-task performance. These same brain regions were not only engaged by a perceptual encoding task in Experiment 2, their activity also tracked delays to a speeded decision-making task caused by concurrent perceptual encoding (Experiment 3). We conclude that a unified attentional bottleneck, including the inferior frontal junction, superior medial frontal cortex, and bilateral insula, temporally limits operations as diverse as perceptual encoding and decision-making.
0 Communities
1 Members
0 Resources
14 MeSH Terms