Method in the preparation of samples for microscopic examination and for checking coverslipping quality
The invention relates to a method in the preparation of samples for microscopic examination onto which a coverslip is applied. The method is notable for the fact that the coverslipping quality is checked automatically and at least partly optically. The invention further relates to an apparatus for carrying out the method, and to an apparatus for checking the coverslipping quality of samples onto which a coverslip is applied.
1. A method of preparing samples for microscopic examination, comprising:
automatically and optically checking quality of a solid coverslip on a specimen slide with dark field illumination when the specimen slide is in a coverslipping apparatus, wherein the solid coverslip is a glass coverslip;
checking a staining quality of a stain applied to the specimen slide using the coverslipping apparatus when the specimen slide is in the coverslipping apparatus;
automatically identifying, based on the dark field illumination, a defective coverslipped slide using an evaluation apparatus, wherein the defective coverslipped slide exhibits at least one of:
damage to the solid coverslip;
fracture in the solid coverslip;
presence of air inclusions in a coverslipping medium;
presence of air inclusions between the solid coverslip and a sample on the specimen slide; and
presence of surface irregularities of the solid coverslip; and
based on results of the evaluation apparatus, re-coverslipping the defective coverslipped slide or excluding the defective coverslipped slide from at least one microscopic examination and providing a non-defective coverslipped slide to a subsequent laboratory process.
2. The method according to claim 1 , further comprising generating an image of at least one of the solid coverslip or the coverslipping medium.
3. The method according to claim 2 , wherein the image is a grayscale image.
4. The method according to claim 2 , wherein the image comprises a microscopic image.
5. The method according to claim 2 , wherein the solid coverslip or the coverslipping medium is illuminated during the generating.
6. The method according to claim 2 , wherein the image is automatically evaluated by the evaluation apparatus.
7. The method according to claim 2 , wherein the image is acquired at an image acquisition angle or an illumination angle, and wherein the image acquisition angle or the illumination angle is adjustable.
8. The method according to claim 2 , further comprising measuring at least one of average brightness or brightness distribution of a reference image of a coverslipped specimen slide without a sample before the checking.
9. The method according to claim 8 , wherein an exposure time for the generating is calculated from at least one of the average brightness or the brightness distribution of the reference image.
10. The method according to claim 9 , further comprising obtaining a brightness of a background of the image, wherein the air inclusions are present between the solid coverslip and the sample on the specimen slide, and wherein reflection of illumination light by the air inclusions produces corresponding regions in the image having a brightness greater than the brightness of the background of the image.
11. The method according to claim 8 , wherein a shading correction of the image is carried out on a basis of at least one of the average brightness or the brightness distribution.
12. The method according to claim 2 , further comprising additionally checking the image with regard to at least one of the staining quality of a hematoxylin-eosin stain or a color intensity.
13. The method according to claim 2 , further comprising generating an image of at least one of the solid coverslip or a coverslipping medium with bright field illumination.
14. The method according to claim 13 , further comprising comparing the image in bright field illumination and an image in dark field illumination with one another.
15. The method according to claim 13 , further comprising checking whether correspondence exists between the image with bright field illumination and an image with dark field illumination with respect to a detected detail present in at least one of the image with bright field illumination or the image with dark field illumination.
16. The method according to claim 1 , further comprising generating an overview image for the checking of the solid coverslip on the specimen slide.
17. The method according to claim 1 , further comprising reading out a barcode attached to the specimen slide.
18. The method according to claim 1 , wherein the checking is sequentially performed under two different conditions or at different testing stations.
19. The method according to claim 1 , further comprising additionally checking at least one of the staining quality of a hematoxylin-eosin stain or a color intensity.
20. The method according to claim 1 , further comprising exchanging data regarding the checking with a higher-level data processing system.