5 - Anal. Chem. 1995, 67,2575-2579 Neurotransmitter Imaging...

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Anal. Chem. 1995, 67,2575-2579 Neurotransmitter Imaging in Living Cells Based on Native Fluorescence Detection Weihong Tan,t Vladimir Parpura,# Philip 0. Haydon,* and Edward S. Yetang**+ Ames Laboratoty- USDOE and Department of Chemistry and Department of Zoology and Genetics, Laboratory of Cellular Signaling, Iowa State University, Ames, Iowa 5001 1 A UV laser-based optical microscope and CCD detection system with high sensitivity has been developed to image neurotransmitters in living cells. We demonstrate the detection of serotonin that has been taken up into indi- vidual living glial cells (astrocytes) based on its native fluorescence. We found that the fluorescence intensity of astrocytes increased by up 10 times after serotonin uptake. The temporal resolution of this detection system at M serotonin is as fast as 50 ms, and the spatial resolution is dilli-action limited. This laser microscope imaging system shows promise for studies of spatial- temporal dynamics of neurotransmitter levels in living neurons and glia. The in vivo monitoring of intracellular molecules is of great biological importance. To better understand developmental biol- ogy, cellular differentiation, physiology, and cell biology, a variety of real-time analytical methodologies have been developed.'-1° These approaches have been applied to analyze a large variety of intracellular species ranging from small ions, such as calcium, to large protein molecules. One particularly interesting and widely studied class of molecules is the neurotransmitters, which are responsible for intercellular communication between neurons.5 As in many areas of biology, advances in neuroscience are often directly liked to the development of new techniques for chemical analysis. The release of neurotransmitter has been studied by electrochemical methods, i.e., microelectrodes2 and electrochemical detection coupled with highly efficient separation techniques such as liquid chromatography (HPLC) and capillary electrophoresis (CE) .3 Even though analysis at the single-cell level and subattomole detection limits have been achieved, several problems exist. One of them is the difficulty in achieving simultaneous spatial and temporal + Ames Laboratory-USDOE and Department of Chemistry. Department of Zoology and Genetics. York, 1989. (1) Herman, B.; Jacobson, K Optical Microscopy for Biology; Wiley Liss: New (2) Garris, P. A; Collins, L. B.; Jones, S. R.; Wightman, R. M. 1. Neurochem. (3) Ewing, Mesaros, J. M.; Gavin, P. F. Anal. Chem. 1994,66,527A-537A (4) Yeung, E. S. Ace. Chem. Res. 1994,27, 409-414. (5) Parpura, V.; Basarsky, T. A; Liu, F.; Jeftinija, K; Jeftinija, S.; Haydon, P. G. (6) Tan, W.; Shi, Z.-Y.; Smith, S.; Bimbaum, D.; Kopelman, R Science 1992, (7) Barber, A J. &p. Biol. 1986, 121, 395-406. (8) Wang, X. F.; Periasamy, Herman, B.; Coleman, D. M. Crit. Rev. Anal. (9) Malgaroli, Tsien, R W. Nature 1993, 357, 134-139. 61, 637-647. Nature 1994, 369, 744-747.
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This note was uploaded on 01/15/2012 for the course CHM 2045 taught by Professor Mitchell during the Fall '07 term at University of Florida.

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5 - Anal. Chem. 1995, 67,2575-2579 Neurotransmitter Imaging...

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