Cytological imaging systems and methods
View Patent ↗The present invention relates to the analysis of specimens. Specifically, the invention relates to methods and apparatus for reviewing specimen slides, including apparatus for holding the slides. The invention also relates to an automatic focusing method for an imaging system and methods for accommodating vibration in the imaging system. In particular, the methods and apparatus may be applied to the automated analysis of cytological specimen slides.
1. An automatic focusing method for an optical system, comprising:
performing an initial coarse focus action along a focal axis at a scan position corresponding to a point on a surface of a slide; and
respectively performing a plurality of subsequent fine focus actions along a plurality of focal axes at a plurality of scan positions corresponding to different points on the slide surface, performing the plurality of subsequent fine focus actions comprising obtaining a plurality of images of the slide at a plurality of coordinates, each image comprising a two-dimensional array of pixels, wherein each of the fine focus actions is performed more quickly than the initial coarse focus action.
2. The method of claim 1 , wherein the performance of the coarse focus action comprises determining a coarse in-focus coordinate along the focal axis.
3. The method of claim 2 , wherein the performance of each of the fine focus actions is based on the coarse in-focus coordinate determined in the coarse focus action.
4. The method of claim 3 , wherein the performance of each of the fine focus actions comprises estimating a fine in-focus coordinate along the respective focal axis as a function of the coarse in-focus coordinate determined in the coarse focus action.
5. The method of claim 3 , wherein the performance of at least one of the fine focus actions comprises estimating a fine in-focus coordinate along the respective focal axis as a function of a global focal plane.
6. The method of claim 1 , wherein the performance of the coarse focus action comprises repeatedly obtaining an image of the slide at different coordinates along the focal axis until a coarse in-focus coordinate is determined.
7. The method of claim 1 , wherein the performance of each of the fine focus actions comprises estimating a fine in-focus coordinate along the respective focal axis and obtaining images of the slide at predetermined coordinates relative to the estimated fine in-focus coordinate along the respective focal axis.
8. The method of claim 1 , wherein the performance of at least one of the coarse focus action and each fine focus action comprises:
obtaining images of the slide at a plurality of coordinates along the focal axis;
determining a plurality of focus scores for the respective coordinates; and
selecting one of the coordinates as an in-focus coordinate based on the focus scores.
9. The method of claim 8 , wherein the coordinate having a maximum focus score is the coordinate selected as the in-focus coordinate.
10. The method of claim 1 , wherein the slide carries a biological specimen.
11. The method of claim 1 , wherein the coarse focus action and fine focus actions are performing during a single image scan.
12. The method of claim 1 , wherein the performance of one or both of the coarse focus action and fine focus actions comprises moving an element of the optical system relative to the slide surface to coordinates along the respective focal axes.
13. The method of claim 1 , wherein the performance of the fine focus actions comprises moving an element of the optical system relative to the slide along a scan axis to the respective scan positions.
14. The method of claim 1 , the two-dimensional array of pixels having a 640×480 pixel array size.
15. The method of claim 1 , further comprising
examining the two-dimensional arrays of pixels of the plurality of images; and
selecting one of the plurality coordinates as a fine in-focus coordinate based on examining the two-dimensional arrays of pixels.
16. The method of claim 15 , examining the two-dimensional arrays comprising comparing pixels to each other.
17. The method of claim 16 , examining the two-dimensional arrays comprising comparing gray scale values of the pixels.
18. The method of claim 17 , comparing gray scale values comprising comparing gray scale values between pairs of pixels separated by a fixed number of pixels.
19. The method of claim 15 , wherein the selected coordinates are evenly distributed within a pre-determined range.
20. The method of claim 1 , performing the initial coarse focus action comprising repeatedly obtaining an image of the slide at different coordinates along the focal axis.
21. The method of claim 1 , wherein performing the initial coarse focus action and performing each of the fine focus actions comprises:
obtaining images of the slide at a plurality of coordinates along the focal axis;
determining a plurality of focus scores for the respective coordinates; and
selecting one of the coordinates as an in-focus coordinate based on the focus scores.
22. The method of claim 5 , the slide comprising at least three fiducial points, the method further comprising determining the global focal plane based on the at least three fiducial points.
23. The method of claim 22 , determining the global focal plane comprising:
determining positions of the at least three fiducial points; and
determining a scan axis slope, an index axis slope and a focus intersect of the global focal plane based on the positions of the at least three fiducial points.