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1.
J Lab Autom ; 21(2): 260-7, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26391011

RESUMO

Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) is a powerful tool for investigating the distribution of proteins and other molecules within biological systems through the in situ analysis of tissue sections, enabling molecular histology. MALDI IMS can determine the distribution of hundreds of unknown compounds in a single measurement while maintaining spatial and molecular integrity. The matrix spraying stage is a key factor in making this technique more sensitive and robust. In this article, we describe a custom-made matrix sprayer (Langartech), which is both inexpensive (estimated cost of about €3000, or $3500) and reliable compared with the alternatives present in the market today, with prices greater than €20,000 ($25,000). Several comparisons were made between our Langartech sprayer and one of the high-end matrix sprayers commercially available: ImagePrep (Bruker Daltonics). Focusing on lateral resolution and observed peak intensities, overall results show that our sprayer behaves in a very competitive fashion, especially when taking into account the huge difference in sophistication level and price.


Assuntos
Imagem Óptica/métodos , Manejo de Espécimes/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Animais , Rim/química , Rim/ultraestrutura , Camundongos , Imagem Óptica/economia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/economia
2.
Sci Rep ; 6: 27106, 2016 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-27256770

RESUMO

Aortic stenosis (AS) is the most common form of valve disease. Once symptoms develop, there is an inexorable deterioration with a poor prognosis; currently there are no therapies capable of modifying disease progression, and aortic valve replacement is the only available treatment. Our goal is to study the progression of calcification by matrix-assisted laser desorption ionization imaging mass spectrometry (MALDI-IMS) and get new insights at molecular level that could help in the understanding of this disease. In this work, we analyzed consecutive slices from aortic valve tissue by MALDI-IMS, to establish the spatial distribution of proteins and peptides directly from the surface of the histological sections. The analysis showed different structures corresponding to regions observed in conventional histology, including large calcification areas and zones rich in collagen and elastic fibers. Peptide extraction from the tissue, followed by liquid chromatography mass spectrometry analysis, provided the identification of collagen VI α-3 and NDRG2 proteins which correlated with the masses obtained by MALDI-IMS and were confirmed by immunohistochemistry. These results highlighted the molecular mechanism implied in AS using MALDI-IMS, a novel technique never used before in this pathology. In addition, we can define specific regions proving a complementary resolution of the molecular histology.


Assuntos
Estenose da Valva Aórtica/genética , Valva Aórtica/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Substituição da Valva Aórtica Transcateter , Idoso de 80 Anos ou mais , Valva Aórtica/metabolismo , Valva Aórtica/patologia , Valva Aórtica/cirurgia , Estenose da Valva Aórtica/metabolismo , Estenose da Valva Aórtica/patologia , Estenose da Valva Aórtica/cirurgia , Biomarcadores/metabolismo , Colágeno Tipo VI/genética , Colágeno Tipo VI/metabolismo , Expressão Gênica , Humanos , Imuno-Histoquímica , Masculino , Microtomia , Inclusão do Tecido , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo
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