Method for examining a specimen, and scanning microscope system
A method for examining a specimen ( 27 ) that exhibits at least two optical transition lines and is optically excitable at least with light of a first and light of a second wavelength is characterized by the step of illuminating the specimen ( 27 ) with illuminating light ( 15 ) that generates at least a multiple of the first wavelength and a multiple of the second wavelength; and by the step of detecting the detected light ( 29 ) proceeding from the specimen ( 27 ). Also disclosed is a scanning microscope system ( 1 ) having at least one light source ( 3 ) that emits illuminating light ( 15 ) for illumination of a specimen ( 27 ), the specimen ( 27 ) exhibiting at least two optical transition lines and being optically excitable at least with light of a first and light of a second wavelength, having at least one detector ( 41, 43, 65, 77, 79 ) for detection of the detected light ( 29 ) proceeding from the specimen ( 27 ) and an objective ( 25 ) for focusing the illuminating light ( 15 ) onto a subregion of the specimen ( 27 ). The scanning microscope system is characterized in that the illuminating light ( 15 ) generates at least a multiple of the first wavelength and a multiple of the second wavelength.
1. A method for examining a specimen that exhibits at least two optical transition lines and is optically excitable with light of at least a first and light of a second wavelength, the method comprises the steps of:
illuminating the specimen with an illuminating light composed of at least a multiple of the first wavelength and a multiple of the second wavelength, wherein a portion of said illuminating light is produced by a microstructured optical element, said microstructured optical element constructed from a plurality of microoptical structural elements having at least two different optical densities that converts light impinging thereon into light of at least one other wavelength; and,
detecting a detection light proceeding from the specimen.
2. The method as defined in claim 1 , wherein a further step comprises focusing the illuminating light onto a subregion of the specimen.
3. The method as defined in claim 1 , wherein a step further comprises:
scanning the specimen by successively guiding the illuminating light over several subregions.
4. The method as defined in claim 1 , wherein the detecting is accomplished confocally.
5. The method as defined in claim 1 , wherein further steps comprise:
converting the detection light into detected signals having an amplitude which depends on a power level of the detection light; and
associating the detected signals with a scanned subregion.
6. The method as defined in claim 1 , wherein at least two detection channels are provided and the detecting is accomplished in as a function of the wavelength of the detected light.
7. The method as defined in claim 1 , wherein the illuminating light is pulsed.
8. The method as defined in claim 1 , wherein the specimen contains an optically excitable marker or a fluorescent dye.
9. The method as defined in claim 1 , wherein the illuminating light is double the first wavelength and double the second wavelength.
10. The method as defined in claim 1 , wherein the illuminating light is triple the first wavelength and triple the second wavelength.
11. The method as defined in claim 1 , wherein the illuminating light contains entangled photons.
12. A scanning microscope system comprising:
a specimen exhibiting at least two optical transition lines and being optically excitable with light of at least a first and light of at least a second wavelength;
at least one light source that emits a light for illumination of the specimen; at least one microstructured optical element that converts a portion of said light to light of at least one other wavelength, said microstructured optical element constructed from a plurality of microoptical structural elements having at least two different optical densities;
at least one detector for detection of the detected light proceeding from the specimen and;
an objective for focusing said light and said light of at least one other wavelength onto a subregion of the specimen, wherein said light and said light of at least one other wavelength comprise at least a multiple of the first wavelength and a multiple of the second wavelength.
13. The scanning microscope system as defined in claim 12 , wherein the light source contains at least one laser that repetitively emits light pulses.
14. The scanning microscope system as defined in claim 13 , wherein the light pulses are shorter in time than 1 picosecond.
15. The scanning microscope system as defined in claim 12 , wherein the microstructured optical element has a first region and a second region, in which context the first region has a homogeneous structure, and a microscopic structure made up of microoptical structural elements is formed in the second region.
16. The scanning microscope system as defined in claim 15 , wherein the first region surrounds the second region.
17. The scanning microscope system as defined in claim 12 , wherein the microstructured optical element is made from glass or plastic material and cavities arranged next to one another.
18. The scanning microscope system as defined in claim 12 , wherein the microoptical structural elements are cannulas, lands, honeycombs, tubes, or cavities.
19. The scanning microscope system as defined in claim 12 , wherein the microstructured optical element is made of photonic band gap material.
20. The scanning microscope system as defined in claim 12 , wherein the microstructured optical element is configured as a light-guiding fiber.
21. The scanning microscope system as defined in claim 20 , wherein the light-guiding fiber has a taper.
22. The scanning microscope system as defined in claim 12 , wherein the illuminating light comprises at least double the first wavelength and at least double the second wavelength.
23. The scanning microscope system as defined in claim 12 , wherein the illuminating light comprises at least triple the first wavelength tripled and at least triple the second wavelength.
24. The scanning microscope system as defined in claim 12 , wherein the detector is a multi-band detector.