Radiant energy – Luminophor irradiation
Patent
1997-07-11
2000-02-01
Hannaher, Constantine
Radiant energy
Luminophor irradiation
2504591, G01N 2164
Patent
active
060205911
ABSTRACT:
A two-photon fluorescence microscope employs two laser beams having pulses of respective wavelengths .lambda..sub.2 and .lambda..sub.3, which cause two-photon emission of a fluorophore when the pulses are spatially and temporally overlapping. The pulses of the two beams of wavelengths .lambda..sub.2 and .lambda..sub.3 are combined at some crossing angle .theta. within the specimen, causing two-photon absorption within a line-shaped region during each instant of overlap. As the pulses pass through each other, the overlapping line-shaped region moves such that a slice of the fluorophore-containing specimen is excited by two-photon absorption during the overlap period. Lateral scanning is effected without moving parts by adjusting the relative delay of the pulses in the two beams. When the crossing angle .theta. is set to 0, i.e., when the two beams are directed along the same axis, the pulses of the two beams form a pancake-shaped volume in which two-photon excitation occurs as the pulses spatially and temporally overlap while traveling in opposite directions. A two-dimensional detector, such as a two-dimensional charge coupled device (CCD) array, can be used to detect a two-dimensional portion of the specimen at one time without lateral scanning. A three-dimensional image can be produced by adjusting the time delay between the two pulses, thereby changing the location of the "pancake" volume created by the intersection of the two pulses.
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Harter Donald J.
Squier Jeff A.
Hannaher Constantine
Imra America Inc.
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