In a previous post it was shown how one could discriminate against broad NMR lines and emphasize sharp NMR lines by using a CPMG T2 filter. This technique has tremendous power for getting detailed information from difficult samples. Biological samples containing in-tact cells can be difficult because they contain semi-solid components with extremely short T2's and liquid components with dissolved compounds having much longer T2's. The CPMG technique was used as early as 1988 to study red blood cells.* The figure below shows 600 MHz 1H NMR data collected at uOttawa on a sample of in-tact red blood cells. The sample, obtained from centrifuged human blood, was paste-like and contained no solvent. The magnet was shimmed using proton gradient shimming and the data were collected without the deuterium lock.
The top panel of the figure shows the result obtained with a standard one-pulse measurement. Clearly, the spectrum shows very little detail and is dominated by the water resonance. The bottom panel shows the CPMG spectrum obtained after 350 echos using an echo delay time (2? = 640 µsec). One can clearly see the suppression of all the broad signals (including the water signal) with short T2's leaving the incredibly detailed and high resolution spectrum of the soluble compounds dissolved in the liquid component of the red blood cells (amino acids, sugars etc...). The water signal is almost completely suppressed.
[NMR paper] Compact NMR relaxometry of human blood and blood components.
Compact NMR relaxometry of human blood and blood components.
Related Articles Compact NMR relaxometry of human blood and blood components.
Trends Analyt Chem. 2016 Nov;83(A):53-64
Authors: Cistola DP, Robinson MD
Abstract
Nuclear magnetic resonance relaxometry is a uniquely practical and versatile implementation of NMR technology. Because it does not depend on chemical shift resolution, it can be performed using low-field compact instruments deployed in atypical settings. Early relaxometry studies of human blood were focused on...
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12-24-2016 08:34 AM
[NMR paper] Robust and low cost uniform (15)N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications.
Robust and low cost uniform (15)N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications.
Robust and low cost uniform (15)N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications.
J Struct Biol. 2014 Aug 27;
Authors: Meola A, Deville C, Jeffers SA, Guardado-Calvo P, Vasiliauskaite I, Sizun C, Girard-Blanc C, Malosse C, Heijenoort CV, Chamot-Rooke J, Krey T, Guittet E, Pêtres S, Rey FA, Bontems F
Abstract
Nuclear...
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09-01-2014 07:46 PM
Robust and low cost uniform 15N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications
Robust and low cost uniform 15N-labeling of proteins expressed in Drosophila S2 cells and Spodoptera frugiperda Sf9 cells for NMR applications
Publication date: Available online 28 August 2014
Source:Journal of Structural Biology</br>
Author(s): Annalisa Meola , Célia Deville , Scott A. Jeffers , Pablo Guardado-Calvo , Ieva Vasiliauskaite , Christina Sizun , Christine Girard-Blanc , Christian Malosse , Carine van Heijenoort , Julia Chamot-Rooke , Thomas Krey , Eric Guittet , Stéphane Pêtres , Félix A. Rey , François Bontems</br>
Nuclear magnetic...
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08-29-2014 05:36 PM
FTIR blood test for liver cancer
FTIR blood test for liver cancer
http://www.spectroscopynow.com/common/images/thumbnails/144d62c377f.jpgFTIR microspectroscopy of human serum can distinguish between healthy livers, cirrhosis of the liver and liver cancer, based on protein structures and lipid levels.
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03-18-2014 09:38 PM
(1)H NMR study of monocrotaline and its metabolites in human blood.
(1)H NMR study of monocrotaline and its metabolites in human blood.
(1)H NMR study of monocrotaline and its metabolites in human blood.
Food Chem Toxicol. 2011 Aug 6;
Authors: Yang YC, Crowder J, Wardle NJ, Yang L, White KN, Wang ZT, Annie Bligh SW
Monocrotaline (MCT) is a naturally occurring hepatotoxic pyrrolizidine alkaloid found in plants. This investigation is aimed at furthering the understanding of the role of blood in mediating the transport of MCT and its reactive metabolites in humans. Reactions of monocrotaline and its metabolites,...
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08-17-2011 01:33 PM
[NMR paper] 1H-NMR investigation of the oxygenation of hemoglobin in intact human red blood cells
1H-NMR investigation of the oxygenation of hemoglobin in intact human red blood cells.
http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--linkinghub.elsevier.com-ihub-images-cellhub.gif http://www.ncbi.nlm.nih.gov/corehtml/query/egifs/http:--www.pubmedcentral.nih.gov-corehtml-pmc-pmcgifs-pubmed-pmc.gif Related Articles 1H-NMR investigation of the oxygenation of hemoglobin in intact human red blood cells.
Biophys J. 1995 Feb;68(2):681-93
Authors: Fetler BK, Simplaceanu V, Ho C
Using improved selective excitation methods for protein nuclear...
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08-22-2010 03:41 AM
Need help in CPMG
I tried to combine the exchange enhanced sensitivity pulse sequence (Wang CY, Grey MJ, Palmer AG. 2001. Journal of Biomolecular Nmr 21: 361-6) with the TROSY detection scheme (Loria JP, Rance M, Palmer AG. 1999. Journal of Biomolecular Nmr 15: 151-5) by implementing Wang's sequences up to the end point of transverse relaxation(without time labeling) followed by Loria's detection sequence. The 15N 180 degree pulse width was limited to 160us by probe.
Applying the relaxation compensated sequence to a 19kD protein results in oscillating(with respect to Echo delay) R2 around 5Hz, while the...