Closed loop monitoring of automated molecular pathology system
View Patent ↗A closed loop automated method for staining of a biological sample is provided. The method comprises providing a biological sample, staining at least a portion of the biological sample by flowing in a reagent, monitoring one or more optical characteristics of the biological sample, and calculating a figure of merit based on at least one of the optical characteristics. An automated device for iterative staining of a biological sample is also provided.
1. A closed loop automated method for staining of a biological sample, comprising:
providing a biological sample;
staining at least a portion of the biological sample by flowing in a reagent;
during the staining, monitoring one or more optical characteristics of the biological sample until saturation of at least one of the one or more optical characteristics;
ceasing the reagent flow after saturation of the at least one of the one or more optical characteristics; and
calculating a figure of merit based on at least one of the one or more optical characteristics.
2. The method of claim 1 , wherein the figure of merit comprises one or more of a light intensity, a contrast of an image, a Brenner gradient, or a signal to background ratio.
3. The method of claim 1 , wherein monitoring comprises acquiring images from the portion of the biological sample during staining.
4. The method of claim 3 , comprising closing an illumination shutter when not imaging.
5. The method of claim 1 , further comprising adding one or more anti-fading agents to the biological sample to reduce any photo-bleaching of the biological sample.
6. The method of claim 1 , further comprising:
destaining at least a portion of the stained portion of the biological sample;
monitoring one or more optical characteristics of the destained portion of the biological sample;
calculating a figure of merit based on at least one of the one or more optical characteristics of the destained portion of the biological sample.
7. The method of claim 6 , wherein monitoring comprises acquiring images from the destained portion of the biological sample during destaining.
8. The method of claim 1 , further comprising:
rinsing at least a portion of the stained portion of the biological sample; and
monitoring one or more optical characteristics of the rinsed portion of the biological sample; and
calculating a figure of merit based on at least one of the optical characteristics of the rinsed portion of the biological sample.
9. The method of claim 8 , wherein monitoring comprises acquiring images from the portion of the biological sample during rinsing.
10. The method of claim 9 , further comprising imaging regions away from the rinsed portion of the biological sample.
11. The method of claim 1 , further comprising acquiring an initial image of the biological sample prior to staining.
12. The method of claim 11 , further comprising rinsing the biological sample with a buffer solution prior to acquiring the initial image.
13. The method of claim 1 , further comprising rinsing at least a portion of the biological sample, destaining at least a portion of the biological sample, and wherein monitoring comprises continuously acquiring images during staining, destaining or rinsing.
14. The method of claim 13 , comprising real-time monitoring of the biological sample.
15. The method of claim 1 , further comprising rinsing at least a portion of the biological sample, destaining at least a portion of the biological sample, and wherein monitoring comprises intermittently acquiring images during staining, destaining or rinsing.
16. The method of claim 15 , wherein intermittently acquiring images comprises acquiring images at time intervals greater than an exposure time of the biological sample to an illumination source.
17. The method of claim 1 , wherein determining the figure of merit comprises correlating light intensity values with specific labeling of a biomarker.