Sample processing improvements for microscopy
Among other things, a first surface is configured to receive a sample and is to be used in a microscopy device. There is a second surface to be moved into a predefined position relative to the first surface to form a sample space that is between the first surface and the second surface and contains at least part of the sample. There is a mechanism configured to move the second surface from an initial position into the predefined position to form the sample space. When the sample is in place on the first surface, the motion of the second surface includes a trajectory that is not solely a linear motion of the second surface towards the first surface.
1. A method comprising:
moving a second surface towards a first surface, on which a fluid specimen is arranged, to form a single layer of sample units in the fluid specimen in at least a portion of a space between the first surface and the second surface, wherein the sample units comprise a light absorbing substance;
illuminating the fluid specimen with light having a first wavelength;
capturing an image of the fluid specimen illuminated with the light having the first wavelength;
identifying, in the image, a first sample unit of the single layer of sample units;
determining, based on the image, a first intensity corresponding to light passing through the first sample unit;
determining, based on the image, a background intensity at a location of the first sample unit;
determining a path length through the single layer of sample units; and
based on the first intensity, the background intensity, the first wavelength, and the path length, determining a concentration of the light absorbing substance.
2. The method of claim 1 , wherein determining the concentration of the light absorbing substance comprises determining a concentration of the light absorbing substance in the first sample unit.
3. The method of claim 1 , wherein the first sample unit comprises a red blood cell, and wherein the light absorbing substance comprises hemoglobin.
4. The method of claim 1 , wherein the path length comprises a distance between the first surface and the second surface.
5. The method of claim 1 , wherein determining the background intensity comprises:
determining, based on the image, a second intensity correspond to light passing through a background medium in a region of the fluid specimen in proximity to the first sample unit without passing through a sample unit; and
determining the background intensity based on the second intensity.
6. The method of claim 5 , wherein determining the background intensity based on the second intensity comprises performing interpolation or extrapolation from the region of the fluid specimen to the location of the first sample unit.
7. The method of claim 1 , wherein moving the second surface towards the first surface comprises moving the second surface to a distance from the first surface that is set by beads in the fluid specimen, and
wherein the path length comprises a diameter of the beads.
8. The method of claim 1 , wherein capturing the image comprises capturing the image using a light-sensitive pixel array, and wherein the method comprises:
arranging the fluid specimen on the first surface within a near-field distance from the pixel array.
9. The method of claim 8 , wherein the near-field distance is less than the first wavelength.
10. The method of claim 1 , wherein determining the first intensity comprise determining an average intensity of light detected by pixels that underlie the first sample unit.
11. The method of claim 1 , wherein moving the second surface towards the first surface comprises repeatedly moving the second surface towards and away from the first surface while the second surface is in contact with the fluid specimen, to cause mixing of the fluid specimen.
12. An apparatus comprising:
a first surface arranged to receive a fluid specimen within a near-field distance of a light-sensitive pixel array;
a second surface and a movement mechanism configured to move the second surface to within a predetermined distance from the first surface, to form a single layer of sample units in the fluid specimen in at least a portion of a space between the first surface and the second surface, wherein the sample units comprise a light absorbing substance;
a light source; and
electronics coupled to the light source and the light-sensitive pixel array, the electronics configured to perform operations comprising:
causing the light source to illuminate the fluid specimen with light having a first wavelength;
causing the light-sensitive pixel array to capture an image of the fluid specimen illuminated with the light having the first wavelength;
identifying, in the image, a first sample unit of the single layer of sample units;
determining, based on the image, a first intensity corresponding to light passing through the first sample unit;
determining, based on the image, a background intensity at a location of the first sample unit;
determining a path length through the single layer of sample units; and
based on the first intensity, the background intensity, the first wavelength, and the path length, determining a concentration of the light absorbing substance.
13. The apparatus of claim 12 , wherein determining the concentration of the light absorbing substance comprises determining a concentration of the light absorbing substance in the first sample unit.
14. The apparatus of claim 12 , wherein the first sample unit comprises a red blood cell, and wherein the light absorbing substance comprises hemoglobin.
15. The apparatus of claim 12 , wherein the path length comprises a distance between the first surface and the second surface.
16. The apparatus of claim 12 , wherein determining the background intensity comprises:
determining, based on the image, a second intensity correspond to light passing through a background medium in a region of the fluid specimen in proximity to the first sample unit without passing through a sample unit; and
determining the background intensity based on the second intensity.
17. The apparatus of claim 16 , wherein determining the background intensity based on the second intensity comprises performing interpolation or extrapolation from the region of the fluid specimen to the location of the first sample unit.
18. The apparatus of claim 12 , wherein the near-field distance is less than the first wavelength.
19. The apparatus of claim 12 , wherein determining the first intensity comprises determining an average intensity of light detected by pixels that underlie the first sample unit.
20. The apparatus of claim 12 , wherein the electronics are configured to control the movement mechanism to repeatedly move the second surface towards and away from the first surface while the second surface is in contact with the fluid specimen, to cause mixing of the fluid specimen.