Home Abstract Introduction
Construction of glutamate sensors Initial testing of sensors in vitro Optimization of sensor characteristics Expression of sensor in living cells Glutamate measurement in dendrites Glutamate measurement in response to frequency of action potentials
Glutamate function Sensor design
Protein purification through His tags Overexpression of protein using pRSETB Directing proteins to a membrane by signal peptides Transmembrane regions of proteins Expression of proteins on the surface of mammalian cells through pDISPLAY

Journal of Comprehensible Results

Hires SA, Zhu Y, Tsien R (2008)
Optical measurement of synaptic glutamate spillover and reuptake
by linker optimized glutamate-sensitive fluorescent reporters
Proc Natl Acad Sci USA 105:4411-4416

(Translated by Jeff Elhai)

Experiment: Expression of sensor in living cells

[Brief explanation of transfection]

[Brief explanation of HEK293 cells]

[image]
[Cartoon of plasmid into cell, regrown to dendrite]
[Membrane bound response in HEK293 cells to known amount of glutamate (100 μM), using original sensor and optimized sensor.]

[Change log scale to linear. Change "0N0C" to "original", "8N5C" to "optimized"]