Unique oblique lighting technique using a brightfield darkfield objective and imaging method relating thereto
A process is provided for imaging a surface of a specimen with an imaging system that employs a BD objective having a darkfield channel and a bright field channel, the BD objective having a circumference. The specimen is obliquely illuminated through the darkfield channel with a first arced illuminating light that obliquely illuminates the specimen through a first arc of the circumference. The first arced illuminating light reflecting off of the surface of the specimen is recorded as a first image of the specimen from the first arced illuminating light reflecting off the surface of the specimen, and a processor generates a 3D topography of the specimen by processing the first image through a topographical imaging technique. Imaging apparatus is also provided as are further process steps for other embodiments.
1. An imaging apparatus for imaging a surface of a specimen, the imaging apparatus comprising:
a microscope selected from reflected light microscopes and transmitted light microscopes, the microscope including:
a brightfield/darkfield (BD) objective having a darkfield channel and a brightfield channel, the BD objective having a circumference; and
a light barrier positioned in a nosepiece of the microscope and outside the BD objective, the light barrier comprising a body that selectively permits passage of light therethrough, the light barrier comprising a darkfield opening that defines a passage for illuminating light traveling through the darkfield channel toward the specimen, such that the illuminating light does not pass through an entire circumference of the BD objective.
2. The imaging apparatus of claim 1 , wherein the darkfield opening defines arced illuminating light that obliquely illuminates the specimen through the darkfield channel from a discrete direction throughout only an arc of a circumference of the BD objective.
3. The imaging apparatus of claim 2 , wherein the nosepiece is coupled with a bearing housing that is secured to the nosepiece of the microscope.
4. The imaging apparatus of claim 3 , wherein the bearing housing comprises a plurality of bearings permitting rotation of the light barrier to position the darkfield opening at a desired position about a circumference of the BD objective.
5. The imaging apparatus of claim 4 , wherein the arc is from 1 degree or more to 180 degrees or less.
6. The imaging apparatus of claim 4 , wherein the arc is from 2 degrees or more to 5 degrees or less.
7. The imaging apparatus of claim 1 , wherein the light barrier does not intrude upon the darkfield channel.
8. The imaging apparatus of claim 1 , further comprising:
a camera configured to record a first image of the specimen from a first arced illuminating light reflecting off the surface of the specimen, wherein the first arced illuminating light reflecting off the surface of the specimen is reflected back through the brightfield channel.
9. The imaging apparatus of claim 8 , further comprising:
a computing device in communication with the camera, the computing device configured to generate a 3D topography of the specimen by processing the first image through a topographical imaging technique, wherein topographical data are generated from the first image.
10. A method for imaging a surface of a specimen with an imaging system that employs a microscope selected from a reflected light microscope and a transmitted light microscope including a brightfield/darkfield (BD) objective having a darkfield channel and a brightfield channel, the BD objective having a circumference, the method comprising:
rotating a light barrier to a first position, the light barrier positioned in a nosepiece of the reflected light microscope and outside the BD objective; and
obliquely illuminating the specimen through the darkfield channel with a first arced illuminating light that obliquely illuminates the specimen through a first arc of the circumference as defined by the light barrier in the first position, said first arced illuminating light reflecting off of the surface of the specimen, by:
delivering illuminating light into the darkfield channel to the light barrier and through a darkfield opening to provide the first arced illuminating light that obliquely illuminates the specimen; and
recording a first image of the specimen from the first arced illuminating light reflecting off the surface of the specimen, wherein the first arced illuminating light reflecting off the surface of the specimen is reflected back through the brightfield channel and recorded as the first image.
11. The method of claim 10 , further comprising:
generating a 3D topography of the specimen by processing the first image through a topographical imaging technique, wherein topographical data is generated from the first image.
12. The method of claim 10 wherein rotating the light barrier positioned in the nosepiece of the reflected light microscope, comprises:
rotating a bearing housing configured to couple the nosepiece to the microscope.
13. The method of claim 12 , wherein the bearing housing comprises a plurality of bearings permitting rotation of the light barrier to position the darkfield opening at a desired position about a circumference of the BD objective.
14. The method of claim 10 , wherein the darkfield opening defines arced illuminating light that obliquely illuminates the specimen through the darkfield channel from a discrete direction throughout only an arc of a circumference of the BD objective.
15. The method of claim 14 wherein rotating the light barrier positioned in the nosepiece of the reflected light microscope, comprises:
rotating the light barrier such that the arc is from 1 degree or more to 180 degrees or less.
16. The method of claim 14 wherein rotating the light barrier positioned in the nosepiece of the reflected light microscope, comprises:
rotating the light barrier such that the arc is from 2 degrees or more to 5 degrees or less.
17. The method of claim 10 , further comprising:
rotating the light barrier to a second position;
obliquely illuminating the specimen through the darkfield channel with a second arced illuminating light that obliquely illuminates the specimen through a second arc of the circumference different from said first arc as defined by the light barrier positioned in the second position, said second arced illuminating light reflecting off of the surface of the specimen; and
recording a second image of the specimen from the second arced illuminating light reflecting off the surface of the specimen, wherein the first arced illuminating light reflecting off the surface of the specimen is reflected back through the brightfield channel and recorded as the second image.
18. A microscope for analyzing a specimen comprising:
a brightfield/darkfield (BD) objective having a darkfield channel and a brightfield channel, the BD objective having a circumference; and
a light barrier positioned in a nosepiece of the microscope and outside the BD objective, the light barrier comprising a body that selectively permits passage of light therethrough, the light barrier comprising a darkfield opening that defines a passage for illuminating light traveling through the darkfield channel toward the specimen, such that the illuminating light does not pass through an entire circumference of the BD objective.
19. The microscope of claim 18 , wherein the nosepiece is coupled with a bearing housing that is secured to the nosepiece of the microscope.
20. The microscope of claim 19 , wherein the bearing housing comprises a plurality of bearings permitting rotation of the light barrier to position the darkfield opening at a desired position about a circumference of the BD objective.