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Use of the 1-mm micro-probe for metabolic analysis on small volume biological samples.

Serkova NJ, Freund AS, Brown JL, Kominsky DJ - J. Cell. Mol. Med. (2009)

Bottom Line: Clinical application of high-resolution NMR spectroscopy is often limited by extremely low volumes of human specimens.In the present study, the use of the Bruker 1-mm high-resolution TXI micro-probe was evaluated in the elucidation of metabolic profiles for three different clinical applications with limited sample sizes (body fluids, isolated cells and tissue biopsies).In this study, the use of the Bruker 1-mm micro-probe provides a convenient way to measure and quantify endogenous metabolic profiles of samples with a very low volume/weight/cell count.

View Article: PubMed Central - PubMed

Affiliation: Biomedical MRI/MRS Cancer Center Core, University of Colorado Health Sciences Center, Denver, CO 80262, USA.

ABSTRACT
Endogenous metabolites are promising diagnostic end-points in cancer research. Clinical application of high-resolution NMR spectroscopy is often limited by extremely low volumes of human specimens. In the present study, the use of the Bruker 1-mm high-resolution TXI micro-probe was evaluated in the elucidation of metabolic profiles for three different clinical applications with limited sample sizes (body fluids, isolated cells and tissue biopsies). Sample preparation and (1)H-NMR metabolite quantification protocols were optimized for following oncology-oriented applications: (i) to validate the absolute concentrations of citrate and spermine in human expressed prostatic specimens (EPS volumes 5 to 10 microl: prostate cancer application); (ii) to establish the metabolic profile of isolated human lymphocytes (total cell count 4 x 10(6): chronic myelogenous leukaemia application); (iii) to assess the metabolic composition of human head-and-neck cancers from mouse xenografts (biopsy weights 20 to 70 mg: anti-cancer treatment application). In this study, the use of the Bruker 1-mm micro-probe provides a convenient way to measure and quantify endogenous metabolic profiles of samples with a very low volume/weight/cell count.

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Mentions: All men were presumed cancer‐free at the time of EPS collection. Using two‐dimensional HSQC and COSY spectra, the peaks for alanine, citrate, myo‐inositol, lactate, phosphocholine, spermine, valine were identified and quantified on one‐dimensional 1H‐NMR spectra (Fig. 2A and B). Specifically, the ‘normal’ EPS profile had pronounced NMR peaks for the amino acid citrate (concentrations range 161.6 to 764.5 μmol/ml), as well as the polyamine spermine (18.9 to 168 μmol/ml) and the osmolyte myo‐inositol (7.7–42 μmol/ml), and diminished peaks for alanine, lactate, phosphocholine and valine. There was some age‐dependency for higher concentration ranges of citrate and spermine corresponding to younger subjects – a tendency which will be confirmed and validated in a large ongoing clinical study.


Use of the 1-mm micro-probe for metabolic analysis on small volume biological samples.

Serkova NJ, Freund AS, Brown JL, Kominsky DJ - J. Cell. Mol. Med. (2009)

© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4940789&req=5

Mentions: All men were presumed cancer‐free at the time of EPS collection. Using two‐dimensional HSQC and COSY spectra, the peaks for alanine, citrate, myo‐inositol, lactate, phosphocholine, spermine, valine were identified and quantified on one‐dimensional 1H‐NMR spectra (Fig. 2A and B). Specifically, the ‘normal’ EPS profile had pronounced NMR peaks for the amino acid citrate (concentrations range 161.6 to 764.5 μmol/ml), as well as the polyamine spermine (18.9 to 168 μmol/ml) and the osmolyte myo‐inositol (7.7–42 μmol/ml), and diminished peaks for alanine, lactate, phosphocholine and valine. There was some age‐dependency for higher concentration ranges of citrate and spermine corresponding to younger subjects – a tendency which will be confirmed and validated in a large ongoing clinical study.

Bottom Line: Clinical application of high-resolution NMR spectroscopy is often limited by extremely low volumes of human specimens.In the present study, the use of the Bruker 1-mm high-resolution TXI micro-probe was evaluated in the elucidation of metabolic profiles for three different clinical applications with limited sample sizes (body fluids, isolated cells and tissue biopsies).In this study, the use of the Bruker 1-mm micro-probe provides a convenient way to measure and quantify endogenous metabolic profiles of samples with a very low volume/weight/cell count.

View Article: PubMed Central - PubMed

Affiliation: Biomedical MRI/MRS Cancer Center Core, University of Colorado Health Sciences Center, Denver, CO 80262, USA.

ABSTRACT
Endogenous metabolites are promising diagnostic end-points in cancer research. Clinical application of high-resolution NMR spectroscopy is often limited by extremely low volumes of human specimens. In the present study, the use of the Bruker 1-mm high-resolution TXI micro-probe was evaluated in the elucidation of metabolic profiles for three different clinical applications with limited sample sizes (body fluids, isolated cells and tissue biopsies). Sample preparation and (1)H-NMR metabolite quantification protocols were optimized for following oncology-oriented applications: (i) to validate the absolute concentrations of citrate and spermine in human expressed prostatic specimens (EPS volumes 5 to 10 microl: prostate cancer application); (ii) to establish the metabolic profile of isolated human lymphocytes (total cell count 4 x 10(6): chronic myelogenous leukaemia application); (iii) to assess the metabolic composition of human head-and-neck cancers from mouse xenografts (biopsy weights 20 to 70 mg: anti-cancer treatment application). In this study, the use of the Bruker 1-mm micro-probe provides a convenient way to measure and quantify endogenous metabolic profiles of samples with a very low volume/weight/cell count.

Show MeSH
Related in: MedlinePlus