Systems and methods for multicolor imaging
Disclosed herein, inter alia, are methods and systems of image analysis useful for rapidly identifying and/or quantifying features.
1. A method of imaging a sample on a sample stage, the method comprising:
moving the sample stage at a sample stage speed;
directing a first excitation beam onto a first location of the sample and detecting a first light emission from the sample with a first sensor array; and
directing a second excitation beam onto a second location of the sample and detecting a second light emission from the sample with a second sensor array;
wherein the first sensor array, the second sensor array, or both the first and the second sensor arrays are configured to generate a charge that transfers across the sensor array at a charge speed associated with the sample stage speed.
2. The method of claim 1 , wherein the sample stage comprises a position encoder.
3. The method of claim 2 , wherein the position encoder generates a synchronization signal that synchronizes the transfer of the charges with the sample stage speed.
4. The method of claim 1 , wherein the first sensor array is configured to generate a first charge that transfers across the first sensor array at a first charge speed, and the second sensor array is configured to generate a second charge that transfers across the second sensor array at a second charge speed.
5. The method of claim 4 , wherein the first charge speed and the second charge speed are synchronized with the sample stage speed.
6. The method of claim 1 , wherein the first location and the second location are separated by about 10 μm, about 20 μm, about 30 μm, about 40 μm, about 50 μm, about 60 μm, about 70 μm, about 80 μm, about 90 μm, or about 100 μm.
7. The method of claim 1 , wherein the first location and the second location are separated by about 40 μm, about 50 μm, or about 60 μm.
8. The method of claim 1 , wherein the first excitation beam and the second excitation beam are both substantially rectangular, comprising a height and width.
9. The method of claim 8 , wherein the first excitation beam and the second excitation beam comprise a height of about 1.0, 1.1 mm, 1.2 mm, 1.3 mm, 1.4 mm, 1.5 mm, 1.6 mm, 1.7 mm, 1.8 mm, 1.9 mm, or 2.0 mm.
10. The method of claim 8 , wherein the first excitation beam and the second excitation beam comprise a width of about 5 μm, 10 μm, 15 μm, 20 μm, 25 μm, or about 30 μm.
11. The method of claim 1 , further comprising detecting a third light emission from the sample with the first sensor array; and detecting fourth light emission from the sample with the second sensor array.
12. The method of claim 1 , comprising moving the sample stage at about 1 mm/second to about 50 mm/second.
13. The method of claim 1 , comprising moving the sample stage at about 30 mm/second or more.
14. The method of claim 1 , wherein each sensor array is at least 8,000 pixels wide and at least 64 pixels long.
15. The method of claim 1 , wherein the first sensor array is about 8,000 pixels wide and about 64 pixels long and the second sensor array is about 8,000 pixels wide and about 128 pixels long.
16. The method of claim 1 , wherein the first light emission and the second light emission comprise diffracted light, refracted light, scattered light, transmitted light, or reflected light.
17. The method of claim 1 , wherein the first light emission and second light emission is a fluorescent emission.
18. The method of claim 1 , further comprising storing a data representation of the sample in a computer readable memory.
19. The method of claim 1 , comprising moving the sample stage in a first direction along a first axis, followed by moving the sample in an opposite direction along the first axis.
20. The method of claim 1 , wherein the sample comprises labeled nucleotides.
21. The method of claim 1 , wherein the sample comprises one or more polynucleotides.
22. The method of claim 1 , wherein the charge speed being associated with the sample stage speed comprises the sample stage speed being a fraction of the charge speed.