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Article: Single-cell imaging tools for brain energy metabolism: a review.

San Martín, Alejandro / Sotelo-Hitschfeld, Tamara / Lerchundi, Rodrigo / Fernández-Moncada, Ignacio / Ceballo, Sebastian / Valdebenito, Rocío / Baeza-Lehnert, Felipe / Alegría, Karin / Contreras-Baeza, Yasna / Garrido-Gerter, Pamela / Romero-Gómez, Ignacio / Barros, L Felipe

Neurophotonics

2014 Volume 1, Issue 1, Page(s) 11004

Abstract: ... resolution characterization of metabolism in the brain and other organs. ... reporters are paving the way toward high-resolution functional mapping of the brain. This review relates ... to a parallel revolution in metabolism. We argue that metabolism needs to be approached both in vitro and ...

Abstract	Neurophotonics comes to light at a time in which advances in microscopy and improved calcium reporters are paving the way toward high-resolution functional mapping of the brain. This review relates to a parallel revolution in metabolism. We argue that metabolism needs to be approached both in vitro and in vivo, and that it does not just exist as a low-level platform but is also a relevant player in information processing. In recent years, genetically encoded fluorescent nanosensors have been introduced to measure glucose, glutamate, ATP, NADH, lactate, and pyruvate in mammalian cells. Reporting relative metabolite levels, absolute concentrations, and metabolic fluxes, these sensors are instrumental for the discovery of new molecular mechanisms. Sensors continue to be developed, which together with a continued improvement in protein expression strategies and new imaging technologies, herald an exciting era of high-resolution characterization of metabolism in the brain and other organs.
Language	English
Publishing date	2014-05-29
Publishing country	United States
Document type	Journal Article
ZDB-ID	2781943-7
ISSN	2329-4248 ; 2329-423X
ISSN (online)	2329-4248
ISSN	2329-423X
DOI	10.1117/1.NPh.1.1.011004
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Thesis ; Online: Single cell analysis and tissue imaging by laser ablation and mass spectrometry

Shrestha, Bindesh

2010

Abstract: ... ionization (LAESI) MS, for metabolomics, tissue imaging and single cell analysis. Chapter 1 introduces ... metabolites and lipids are identified in mouse brain sections using MS with AP IR-MALDI and LAESI ion ... The direct analysis of biochemicals in tissues and single cells is critical for understanding ...

Abstract	The direct analysis of biochemicals in tissues and single cells is critical for understanding living organisms. Due to excellent selectivity and sensitivity, mass spectrometry (MS) has proven to be an invaluable tool for the analysis of the biomolecules. Recent developments in atmospheric pressure direct ionization sources have enabled the in situ analysis of biological samples without external influences (e.g., purification, extraction, matrix addition etc.) that might alter their biochemical makeup. The work presented in this dissertation shows my efforts to utilize two novel atmospheric pressure (AP) direct ionization methodologies, AP infrared (IR) matrix-assisted laser desorption ionization (MALDI) and laser ablation electrospray ionization (LAESI) MS, for metabolomics, tissue imaging and single cell analysis. Chapter 1 introduces analytical techniques used for the analysis of tissues and single cells. The fundamental aspects of IR laser ablation and its utilization in two direct ionization techniques, AP IR-MALDI and LAESI, are reviewed. Chapter 2 introduces AP IR-MALDI for MS. It presents proof-of-principle molecular imaging of mock peptide samples at atmospheric pressure. The utility of AP IR-MALDI for plant tissue imaging and metabolomics are also discussed. Chapter 3 describes the AP IR-MALDI analysis of various pharmaceuticals directly in their commercial formulations, as well as endogenous metabolites, exogenous drug metabolites and synthetic polymers in urine. Chapter 4 presents the application of mid-infrared laser ablation for molecular imaging. The dynamics of the ablation plume and ion production in AP IR-MALDI and LAESI are compared. In Chapter 5 metabolites and lipids are identified in mouse brain sections using MS with AP IR-MALDI and LAESI ion production. Reactive LAESI relies on interactions between the laser ablated species and reactants, e.g., Li +, introduced through the electrospray. This new modality of LAESI enables the analysis of samples with otherwise low ion yields. Chapter 6 discusses the metabolic analysis of single cells by MS at atmospheric pressure. This breakthrough is made possible by the tight focusing of mid-IR laser light through an etched optical fiber tip and accurate aiming of cells for ablation through visualization and micromanipulation. Similar to conventional LAESI, the ablated plume is postionized by an electrospray. Chapter 7 surveys the major challenges in the field of atmospheric pressure ion production based on mid-IR laser ablation. The need for the analysis of smaller cells, reactive LAESI-MS, ultrahigh resolution LAESI-MS, and the potential application of LAESI-MS in laser surgery are discussed.
Keywords	Analytical chemistry
Subject code	621
Language	ENG
Publishing date	2010-01-01 00:00:01.0
Publisher	The George Washington University
Publishing country	us
Document type	Thesis ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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