Anal. Chem. 1995,
Neurotransmitter Imaging in Living Cells Based
Native Fluorescence Detection
Weihong Tan,t Vladimir Parpura,# Philip
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
laser-based optical microscope and CCD detection
sensitivity has been developed
living cells. We demonstrate the
detection of serotonin that has been taken up
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
M serotonin is as fast as
50 ms, and the spatial
resolution is dilli-action
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
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)
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.
Herman, B.; Jacobson, K
Optical Microscopy for Biology;
Wiley Liss: New
Collins, L. B.; Jones,
R.; Wightman, R. M.
Mesaros, J. M.; Gavin, P. F.
Yeung, E. S.
Ace. Chem. Res.
Liu, F.; Jeftinija, K; Jeftinija, S.; Haydon, P.
Tan, W.; Shi, Z.-Y.; Smith,
Bimbaum, D.; Kopelman, R
Wang, X. F.; Periasamy,
Herman, B.; Coleman, D. M.
Tsien, R W.