Standardizing fluorescence microscopy systems
View Patent ↗Systems and methods for standardizing one or more fluorescence scanning instruments to a reference system by separating the effects of drift and normalization. In an embodiment, a drift image comprising an image of a drift reference slide is captured by a system to be standardized. A drift measurement is calculated using the drift image. A first normalization image comprising an image of a normalization slide is also captured by the system to be standardized. A reference normalization image, also comprising an image of the normalization slide, is captured by a reference system. The first normalization image is compared to the reference normalization image to determine a gamma value and offset value for the system to be standardized.
1. A method for standardizing a fluorescence microscopy imaging system, the method comprising:
by a first imaging system to be standardized, capturing a first normalization image comprising an image of a normalization slide;
by a reference imaging system, capturing a reference normalization image comprising an image of the normalization slide;
comparing the first normalization image to the reference normalization image; and
determining a gamma value and offset value for the first imaging system based on the comparison.
2. The method of claim 1 , wherein capturing a first normalization image, comparing the first normalization image to the reference normalization image, and determining a gamma value and offset value for the first imaging system are all performed for each of a plurality of normalization slides.
3. The method of claim 1 , wherein the first normalization image and the reference normalization image comprise images of an identical region of the normalization slide.
4. The method of claim 1 , further comprising, for the reference imaging system, setting a gamma value to one and an offset value to zero.
5. The method of claim 1 , further comprising storing the reference normalization image.
6. The method of claim 1 , further comprising by the first imaging system to be standardized,
capturing a drift image comprising an image of a drift reference slide; and
calculating a drift measurement using the drift image.
7. The method of claim 6 , wherein calculating the drift measurement comprises calculating a ratio of average pixel intensity in the drift image to an exposure of the drift image.
8. The method of claim 6 , further comprising:
by the first imaging system, capturing a digital image of a sample, wherein the digital image comprises a plurality of pixels, each pixel associated with a pixel intensity value; and
altering one or more of the pixel intensity values based on the drift measurement, the gamma value, and the offset value.
9. The method of claim 8 , further comprising storing the digital image without alteration of the pixel intensity values.
10. The method of claim 9 , further comprising applying image processing to the digital image, wherein the alteration of one or more pixel intensity values is performed at a time that the application of the image processing is performed.
11. The method of claim 9 , further comprising displaying the digital image, wherein the alteration of one or more pixel intensity values is performed at a time that the digital image is displayed.
12. The method of claim 6 , further comprising by the reference imaging system, capturing a reference drift image comprising an image of a reference drift slide, and calculating a reference drift measurement using the reference drift image.
13. The method of claim 12 , further comprising storing the reference drift measurement.
14. A system for standardizing a fluorescence microscopy imaging system, the system comprising:
at least one hardware processor; and
at least one executable module that, when executed by the at least one hardware processor,
receives a first normalization image comprising an image of a normalization slide captured by the first imaging system,
receives a reference normalization image comprising an image of the normalization slide captured by a reference imaging system,
compares the first normalization image to the reference normalization image, and
determines a gamma value and offset value for the first imaging system based on the comparison.
15. The system of claim 14 , wherein the first normalization image and the reference normalization image comprise images of an identical region of the normalization slide.
16. The system of claim 14 , wherein at least one executable module, when executed by the at least one hardware processor,
receives a drift image comprising an image of a drift reference slide captured by a first imaging system, and
calculates a drift measurement using the drift image.
17. The system of claim 16 , wherein calculating the drift measurement comprises calculating a ratio of average pixel intensity in the drift image to an exposure of the drift image.
18. The system of claim 16 , wherein the at least one executable module further:
receives a digital image of a sample captured by the first imaging system, wherein the digital image comprises a plurality of pixels, each pixel associated with a pixel intensity value; and
alters one or more of the pixel intensity values based on the drift measurement, the gamma value, and the offset value.
19. A non-transitory computer-readable medium having instructions stored thereon, wherein the instructions, when executed by a processor, cause the processor to:
receive a first normalization image comprising an image of a normalization slide captured by a first imaging system;
receive a reference normalization image comprising an image of the normalization slide captured by a reference imaging system;
compare the first normalization image to the reference normalization image; and
determine a gamma value and offset value for the first imaging system based on the comparison.
20. The non-transitory computer-readable medium of claim 19 , wherein the first normalization image and the reference normalization image comprise images of an identical region of the normalization slide.
21. The non-transitory computer-readable medium of claim 19 , wherein the instructions, when executed by the processor, cause the processor to:
receive a drift image comprising an image of a drift reference slide captured by a first imaging system; and
calculate a drift measurement using the drift image.
22. The non-transitory computer-readable medium of claim 21 , wherein calculating the drift measurement comprises calculating a ratio of average pixel intensity in the drift image to an exposure of the drift image.
23. The non-transitory computer-readable medium of claim 21 , wherein the instructions further cause the processor to:
receive a digital image of a sample captured by the first imaging system, wherein the digital image comprises a plurality of pixels, each pixel associated with a pixel intensity value; and
alter one or more of the pixel intensity values based on the drift measurement, the gamma value, and the offset value.