Apparatus and methods for transmitting light
An apparatus and method for imaging includes an imaging system formed of a movable objective stage proximal to a sample and positioned for providing an excitation beam onto and for capturing an emission from the sample. The movable objective stage includes an optical lens apparatus and a turn reflector optically coupled to the imaging optics, where at least one of the optical lens apparatus and the turn reflector are movable relative to one another for scanning the sample, and wherein the movement is achieved while maintaining a substantially fixed optical path length between the optical lens apparatus and a fixed plane in a fixed imaging optics stage.
1. An apparatus, comprising:
an imaging system having an excitation source for generating an excitation beam, a fixed imaging optics stage composed of a sensor for measuring an emission from a sample, and imaging optics for imaging the emission from the sample onto the sensor; and
a movable objective stage proximal to the sample and positioned for providing the excitation beam onto the sample and for capturing the emission from the sample, where the movable objective stage includes an optical lens apparatus and a turn reflector optically coupled to the imaging optics of the fixed imaging optics stage, and where at least one of the optical lens apparatus and the turn reflector of the movable objective stage are movable relative to one another for scanning of the sample, while maintaining a fixed optical path length between the optical lens apparatus and a fixed plane in the fixed imaging optics stage during movement.
2. The apparatus of claim 1 , wherein the movable objective stage is movable in two orthogonal directions to maintain a fixed optical path length.
3. The apparatus of claim 1 , wherein the excitation source comprises a first excitation source producing a first excitation at a first sampling wavelength that elicits a first sample emission range of wavelengths and a second excitation source producing a second excitation at a second sampling wavelength that elicits a second sample emission range of wavelengths, each of the first excitation, first emission, second excitation, and second emission having a respective optical path.
4. The apparatus of claim 3 , further comprising a compensation plate positioned in one of the respective optical paths.
5. The apparatus of claim 3 , further comprising a compensation plate positioned in a plurality of the respective optical paths.
6. The apparatus of claim 1 , wherein both the optical lens apparatus and the turn reflector of the movable objective stage are movable relative to one another for scanning of a sample area.
7. The apparatus of claim 1 , wherein at least one of the optical lens apparatus and the turn reflector of the movable objective stage are movable relative to one another for scanning multiple samples areas at different positions.
8. The apparatus of claim 7 , further comprising a controller configured to move the at least one of the optical lens apparatus and the turn reflector of the movable objective stage while maintaining the fixed optical path length to sample at the different positions.
9. The apparatus of claim 8 , wherein the controller is configured to continuously move the optical lens apparatus and the turn reflector of the movable objective stage between the different positions.
10. The apparatus of claim 1 , further comprising a controller configured to continuously control movement of the turn reflector during capture of the emission beam from the sample to compensate for vibrational effects during capture.
11. The apparatus of claim 1 , further comprising a controller configured to continuously control movement of the optical lens apparatus and the turn reflector during capture of the emission beam from the sample to compensate for vibrational effects during capture.
12. The apparatus of claim 11 , wherein the controller is configured to continuously control movement of the optical lens apparatus and the turn reflector at different movement increments.
13. The apparatus of claim 1 , further having a controller configured to move the movable objective stage to achieve the fixed optical path length at each of the different sample positions.
14. The apparatus of claim 1 , further comprising a z-stage adjustment controller to adjust a distance between the optical lens apparatus and the sample.
15. The apparatus of claim 1 , wherein the fixed imaging optics stage, the optical lens apparatus, and the turn reflector form a relay lens assembly for imaging the emission into the sensor.
16. The apparatus of claim 1 , wherein the fixed imaging optics stage, the optical lens apparatus, and the turn reflector form an infinite conjugate lens assembly or near infinite conjugate lens assembly.
17. The apparatus of claim 1 , wherein the fixed imaging optics stage and the optical lens apparatus with the turn reflector each form a finite conjugate lens assembly.
18. The apparatus of claim 1 , comprising one or more color separating elements between the movable objective stage and the fixed imaging optics to direct light of a first emission wavelength to a first image sensor and light of a second emission wavelength to a second image sensor.
19. The apparatus of claim 1 , wherein the movable objective stage is separately movable along two orthogonal axes each substantially planar to the sample.
20. The apparatus of claim 1 , comprising one or more color separating elements within or after the fixed imaging optics to direct light of a first emission wavelength to a first image sensor and light of a second emission wavelength to a second image sensor.
21. The apparatus of claim 20 , further comprising a compensating plate disposed before a first image sensor.
22. The apparatus of claim 20 , further comprising a plurality of compensating plates disposed before a first image sensor.
23. The apparatus of claim 20 , further comprising a plurality of compensating plates disposed before a first image sensor and a different compensation plate or different plurality of compensating plates disposed before a second image sensor.
24. The apparatus of claim 21 , wherein one or more compensating plates is tilted or wedged.
25. The apparatus of claim 1 , wherein the movable objective stage is separately movable along two orthogonal axes each substantially parallel to the sample.
26. An apparatus, comprising:
an imaging system having an excitation source for generating an excitation beam, a fixed imaging optics stage composed of a sensor for measuring an emission from a sample, and imaging optics for imaging the emission from the sample onto the sensor; and
an objective stage proximal to the sample and positioned for providing the excitation beam onto the sample and for capturing the emission from the sample, where the objective stage includes an optical lens apparatus,
wherein the imaging system comprises
(i) one or more color separating elements between the objective stage and the fixed imaging optics to direct light of a first emission wavelength to a first image sensor of the sensor and light of a second emission wavelength to a second image sensor of the sensor, or
(ii) the one or more color separating elements within or after the fixed imaging optics to direct light of the first emission wavelength to the first image sensor and light of the second emission wavelength to the second image sensor.
27. The apparatus of claim 26 , further comprising a compensating plate disposed before a first image sensor.
28. The apparatus of claim 26 , wherein a plurality of compensating plates are disposed before a first image sensor.
29. The apparatus of claim 28 , wherein a plurality of compensating plates is disposed before a first image sensor and a different plurality of compensating plates is disposed before a second image sensor.
30. The apparatus of claim 26 , wherein one or more compensating plates is (are) tilted or wedged.
31. The apparatus of claim 26 , further comprising a compensating plate pair disposed within a beam path defined by the one or more color separating elements.
32. The apparatus of claim 31 , wherein the compensating plate pair comprises a first compensating plate tilted in a first angular direction and a second compensating plate tilted in a second angular direction, equal and opposite to the first angular direction.
33. The apparatus of claim 32 , wherein the one or more color separating elements are tilted about a first axis and the first compensating plate and the second compensating plate are each tilted about a second axis orthogonal to the first axis and to the optical axis.
34. A computer-implemented method of optically probing a sample, the method comprising:
aligning, using one or more processors, a movable objective stage, having an optical lens apparatus and a turn reflector optically coupled to imaging optics of a fixed imaging optics stage, to align the optical lens apparatus with the sample for probing at an optical path length;
providing, using the optical lens apparatus, an excitation beam to the sample and capturing, using the optical lens apparatus, a fluorescence emission from the sample;
in response to identification of a shift in focus at the sample from the fluorescence emission, adjusting a position of the optical lens apparatus or a position of the turn reflector to compensate for the shift; and
moving, using the one or more processors, the optical lens apparatus and the turn reflector to position the optical lens apparatus to over a subsequent sample for probing, while maintaining the optical path length.
35. The computer-implemented method of claim 34 , further comprising:
moving, using the one or more processors, the optical lens apparatus and the turn reflector to position the optical lens apparatus to over the subsequent sample for probing while maintaining the optical path length throughout the movement from the sample to the subsequent sample.
36. The computer-implemented method of claim 34 , further comprising:
performing imaging processing on image data containing the fluorescence emission; and
in responding to determining the image data does not satisfy a focusing condition, adjusting a vertical distance between the optical lens apparatus and the sample until the image data satisfies the focusing condition.
37. The computer-implemented method of claim 34 , wherein moving the optical lens apparatus and the turn reflector to position the optical lens apparatus to over the subsequent sample for probing, while maintaining the optical path length comprises moving the optical lens apparatus and the turn reflector in a plane substantially parallel to a plane containing the sample and the subsequent sample.