Microscope with heightened resolution and linear scanning

Abstract

Microscope with heightened resolution and linear scanning wherein the sample is illuminated with a first and a second illuminating light, whereby the first illuminating light excites the sample, and the second illuminating light is generated through the refraction of coherent light at a periodic structure and displays a periodic structure in a lateral beam direction and in axial beam direction.

1. Scanning optical microscope with heightened resolution, comprising:
 first means for generating a first illumination light for illuminating and exciting fluorescence in a sample, 
 second means for generating a second illumination light for illuminating the sample, the second illumination light having a spatially periodic structure in the sample in a lateral beam direction and in an axial beam direction, wherein the second means generates the second illumination light by interference of at least three coherent, partial beams in the sample. 
 
     
     
       2. Microscope according to  claim 1 , wherein the microscope is a light grid microscope with linear scanning. 
     
     
       3. Microscope according to  claim 1 , further comprising means for spatially superimposing patterns created by the first and second illumination lights in the sample, wherein a return of an excited state takes place in the sample in the regions of the second illumination light pattern. 
     
     
       4. Microscope according to  claim 1 , wherein the first and the second illumination lights are laser beams. 
     
     
       5. Microscope according to  claim 1 , further comprising means for moving the first and second illumination lights jointly over the sample. 
     
     
       6. Microscope according to  claim 1 , wherein the first and second illumination lights are linear. 
     
     
       7. Microscope according to  claim 1 , wherein the second means includes a periodic structure that generates the second illumination light through coherent superimposition of several orders of refraction. 
     
     
       8. Microscope according to  claim 7 , wherein the second means further includes a grid for generating the orders of refraction. 
     
     
       9. Microscope according to  claim 8 , wherein the grid generates a Talbot grid effect. 
     
     
       10. Microscope according to  claim 9 , further comprising a microscope objective lens having a depth resolution in the range of the distance between the Talbot planes. 
     
     
       11. Microscope according to  claim 1 , wherein at least one of the first means and the second means includes at least one pulsed laser for generating at least one of the first and second illumination lights. 
     
     
       12. Method of operating a microscope according to  claim 1 , comprising generating a depopulation and an excitation in an alternating manner in the sample. 
     
     
       13. Method according to  claim 12 , wherein the excitation takes place following the depopulation. 
     
     
       14. Method according to  claim 12 , wherein the depopulation takes place following the excitation. 
     
     
       15. Method according to  claim 12 , wherein the first and second means of the microscope includes two pulsed lasers for the generation of the first and the second illumination lights, and wherein the method includes the step of operating the two pulsed lasers in a synchronized matter to cause the excitation and the depopulation to take place alternatingly in the sample. 
     
     
       16. Method according to  claim 12 , further comprising the step of adjusting the optical resolution of the microscope. 
     
     
       17. Method according to  claim 12 , wherein the second means includes a periodic structure that generates the second illumination light through refraction of coherent light, the method further including the step of varying the frequency of the periodic structure to adjust the optical resolution. 
     
     
       18. Method according to  claim 12 , wherein the microscope includes a microscope objective lens and the second means includes a periodic structure that generates the second illumination light through refraction of coherent light, and wherein the method further includes the step of switching of the periodic structure upon switching of the microscope objective lens. 
     
     
       19. Method according to  claim 12 , wherein the second means includes a periodic structure that generates the second illumination light through refraction of coherent light, and the periodic structure is a grid, and wherein the method further comprises the step of switching the grid in the second illumination light. 
     
     
       20. Method for studying developmental processes, comprising the step of: studying dynamic processes in a range of tenths of a second up to hours, on a level of cell associations and entire organisms, using a light grid microscope according to  claim 1 . 
     
     
       21. Method for studying intercellular transport events, comprising the step of: representing small motile structures with high speed, using a light grid microscope according to  claim 1 . 
     
     
       22. Method for representing rapid signal transmission events, comprising the step of: representing neurophysiologic events with high temporal resolution in studies in a muscle and nerve system, using a light grid microscope according to  claim 1 . 
     
     
       23. Scanning optical microscope with heightened resolution, comprising:
 first means for generating a line-shaped first illumination light for illuminating and exciting fluorescence in a sample, 
 second means for generating a line-shaped second illumination light for illuminating the sample, the second illumination light having a spatially periodic structure in the sample in a lateral beam direction and in an axial beam direction, wherein the lateral beam direction is oriented longitudinally with respect to the line-shape of the second illumination light, and wherein the second means generates the second illumination light by interference of at least three coherent, partial beams in the sample. 
 
     
     
       24. Scanning optical microscope with heightened resolution, comprising:
 means for generating a line-shaped illumination light for illuminating a sample by interference of at least three coherent, partial beams in the sample, the illumination light having a spatially periodic structure in the sample in a lateral beam direction and in an axial beam direction, wherein the lateral beam direction is oriented longitudinally with respect to the line-shape of the illumination light;   optics for imaging the illumination light onto the sample to select a region of interest; and   a rotation device for rotating the illumination light relative to the sample to rotate the region of interest.   
     
     
       25. Microscope according to claim 24, wherein the means for generating an illumination light includes a laser light source and a periodic structure illuminated by the laser light source, wherein the periodic structure generates the illumination light through coherent superimposition of several orders of refraction. 
     
     
       26. Microscope according to claim 25, wherein the periodic structure comprises a grid for generating the orders of refraction. 
     
     
       27. Microscope according to claim 24, wherein the rotation device is an Abbe-König prism. 
     
     
       28. Microscope according to claim 24, further comprising means for scanning the illumination light over the sample. 
     
     
       29. Microscope according to claim 24, wherein the spatially periodic structure of the illumination light suppresses at least a portion of excitation fluorescence emitted by the sample.