Flexible light sheet generation by field synthesis
View Patent ↗A light sheet microscopy apparatus has a light source, a spatial light filter, a scanning device positioned between the light source and the spatial light filter, an imaging device, a computer readable medium, and a computer processor coupled to the light source, the scanning device, the imaging device, and the computer-readable medium. The computer processor is configured to operate the light source to generate a light beam, control the scanning device to scan the light beam across the spatial light filter to generate the light sheet, and utilize the imaging device to capture one or more images of light emitted by a sample illuminated by the one or more light sheets.
1. A light sheet microscopy apparatus comprising:
one or more light sources configured to generate one or more light beams;
one or more spatial light filters;
one or more scanning devices positioned between the one or more light sources and the one or more spatial light filters, the one or more scanning devices being controllable to scan the one or more light beams across the one or more spatial light filters, a positive light sheet and a negative light being synthesized by incoherent superposition of one-dimensional intensity distributions from the one or more light beams scanned across the one or more spatial light filters; and
one or more imaging devices configured to capture one or more images of light emitted by a sample illuminated by a final light sheet that is a combination of the positive light sheet and the negative light sheet.
2. The apparatus of claim 1 , wherein the one or more light sources includes one or more lasers.
3. The apparatus of claim 1 , wherein the one or more scanning devices includes one or more galvanometric mirrors.
4. The apparatus of claim 1 , wherein the one or more spatial light filters includes a pupil filter conjugate to a back focal plane.
5. The apparatus of claim 1 , wherein the one or more imaging devices includes one or more cameras.
6. The apparatus of claim 1 ,
wherein,
the one or more scanning devices are moved in one dimension in a first plane parallel to a second plane, and
the second plane contains a principal impingement surface of the one or more spatial light filters.
7. The apparatus of claim 1 , wherein one or more focused lines are scanned across the one or more spatial light filters.
8. The apparatus of claim 1 , wherein the one or more imaging devices are configured to capture sequential images of the light emitted by the sample.
9. The apparatus of claim 1 ,
wherein,
the one or more imaging devices are configured to acquire fluorescence light of different wavelength ranges while the sample is illuminated by the one or more light sheets, and
the one or more light sheets have different wavelengths.
10. A method for a light sheet microscopy, the method comprising:
generating one or more light beams using one or more light sources;
scanning the one or more light beams across one or more spatial filters using one or more scanning devices disposed between the one or more light sources and the one or more spatial filters;
synthesizing a plurality of light sheets by incoherent superposition of intensity distributions varying in at least one dimension from the one or more light beams being scanned across the one or more spatial filters, the plurality of light sheets including a positive light sheet and a negative light sheet;
combining the positive light sheet and the negative light sheet into a final light sheet; and
capturing one or more images of light emitted by a sample illuminated by the final light sheet, the one or more images of light being captured using one or more imaging devices.
11. The method of claim 10 , wherein the one or more light sources includes one or more lasers.
12. The method of claim 10 , wherein the one or more scanning devices includes one or more galvanometric mirrors.
13. The method of claim 10 , wherein the one or more spatial filters includes at least one of a pupil filter or a slit mask.
14. The method of claim 10 , wherein the one or more imaging devices includes one or more cameras.
15. The method of claim 10 ,
wherein,
the scanning of the one or more light beams across the one or more spatial filters includes moving the one or more scanning devices in one dimension in a first plane parallel to a second plane, and
the second plane contains a principal impingement surface of the one or more spatial filters.
16. The method of claim 10 , wherein the scanning of the one or more light beams across the one or more spatial filters includes scanning one or more focused lines across the one or more spatial filters.
17. The method of claim 10 , wherein the capturing of the one or more images of light includes capturing sequential images of light emitted by the sample.
18. The method of claim 17 ,
wherein,
the capturing of the one or more images of light includes acquiring fluorescence light of different wavelength ranges while the sample is illuminated by the one or more light sheets, and
the one or more light sheets have different wavelengths.
19. The method of claim 10 , further comprising:
applying a phase shift to at least a portion of the one or more light beams scanned across the one or more spatial filters to generate the negative light sheet.
20. The method of claim 10 , further comprising:
tuning one or more characteristics of the plurality of light sheets by controlling the one or more light beams being scanned across the one or more spatial filters.
21. A non-transitory computer-readable medium having instructions recorded thereon that, when executed by one or more computer processors, cause the one or more computer processors to perform operations, the operations comprising:
operating one or more light sources to generate one or more light beams;
controlling one or more scanning devices disposed between the one or more light sources and one or more spatial light filters, the one or more scanning devices being controlled to scan the one or more light beams across the one or more spatial light filters, a positive light sheet and a negative light being synthesized by incoherent superposition of intensity distributions varying in a primary direction from the one or more light beams being scanned across the one or more spatial filters; and
capturing one or more images of light emitted by a sample illuminated by a final light sheet that is a combination of the positive light sheet and the negative light sheet, the one or more images of light being captured using one or more imaging devices.
22. A non-transitory computer-readable medium having instructions recorded thereon that, when executed by one or more computer processors, cause the one or more computer processors to perform operations, the operations comprising:
generating one or more images of light emitted by a sample illuminated by a final light sheet that is a combination of a positive light sheet and a negative light sheet, the positive light sheet and the negative light sheet being synthesized by incoherent superposition of one-dimensional intensity distributions from one or more light beams being scanned across one or more spatial filters by controlling one or more scanning devices, the one or more scanning devices disposed between one or more light sources generating the one or more light beams and the one or more spatial filters.
23. The apparatus of claim 1 , wherein the positive light sheet and the negative light sheet are synthesized by the incoherent superposition of one-dimensional intensity distributions from the one or more light beams scanned across the one or more spatial light filters conjugate to a back focal plane.