Laser Microscope At Cornell Images Serotonin In Live Cells

g re-uptake of serotonin into cells from which secretion occurs.

The Cornell technology may be useful in gaining an understanding of a broad spectrum of physiological and psychological effects of this -- and other -- neurotransmitters. The technology could be useful, then, for researchers in designing more effective drugs for a host of disorders.

Previous efforts to image neurotransmitter secretory granules have not directly detected the neurotransmitter content or allowed visualization of secretory processes. But the Cornell technology, called non-linear laser scanning microscopy, can detect and image the serotonin and measure its concentration and the total neurotransmitter content of individual granules in intact cells.

Here is how it works: A laser in the 700 to 750 nanometer wavelength (infrared) fires photons bunched in very short pulses (each 10-13 seconds or a 10 millionth of a millionth of a second), which are focused by the microscope so that there is a high probability that three photons arrive at the same time (in about 10-16 seconds) at the same molecule near the focus. Molecules such as serotonin and tryptophan, which can normally be excited only with deep ultraviolet (~250nm) illumination, are now excited by simultaneously absorbing three infrared photons and, subsequently, fluorescence in the UV. These photons are collected as the laser is scanned through the specimen, and the resulting 3-D image can be viewed and analyzed on a computer monitor. For these studies, the researchers used basal leukemia cells from rats.

"This three-photon excitation produces very high energy corresponding to shorter absorption wavelengths than was possible before, without killing the cells," Webb said. "All you could see before this technique was diffuse brightness increase attributable to serotonin. But now we can see individual granules and we have a way to measure the serotonin in active cells."

Maiti, a postdoctoral associate, said that seroton

Contact: Larry Bernard
Cornell University News Service

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