Optical platelet counter method
Platelets or blood cells are detected in a fluid sample by adjusting a focal depth of a microscope through a range of values, the microscope having a mounted sample and an objective lens adapted with one or both of (a) a spherical aberration correction unmatched to a utilized cover plate for the sample, or (2) a numerical aperture unmatched to a utilized illumination source for the sample. Images are recorded at different specific focal depths and in multiple z planes of a fluid bearing the platelets, where the position of platelets may overlap on different of the multiple z planes that are recorded, the images recorded through the cover plate, thus causing the generation of a specific light-dark pattern indicative of platelets at particular positions and at multiple depths in the fluid media. The images are analyzed for the specific light-dark pattern.
1. An optical organic particle counting method, comprising:
adjusting a microscope focal depth of a microscope through a range of values of the microscope focal depth, the microscope having a microscope focal arrangement adapted to form a plurality of blurry images of organic particles in a z plane of a fluid sample at different ones of the range of values of the microscope focal depth, wherein the different ones of the range of values of the microscope focal depth comprises at least one focal plane above the z plane of the organic particles and at least one focal plane below the z plane of the organic particles;
recording the plurality of blurry images of the organic particles at the different ones of the range of values of the microscope focal depth, wherein coordinates of the organic particles overlap on the plurality of blurry images, thus causing the generation of a specific light-dark pattern across the plurality of blurry images indicative of the organic particles; and
analyzing the plurality of blurry images for the specific light-dark pattern across the plurality of blurry images to identify and classify the organic particles.
2. The method of claim 1 , further comprising:
the range of values of the microscope focal depth being approximately 320 micrometers and a slide depth of approximately 100 micrometers.
3. The method of claim 1 , further comprising:
the plurality of blurry images being at least 40 images recorded at three images per second.
4. The method of claim 1 , further comprising:
the microscope focal arrangement comprising a blue LED illumination source having a wavelength of approximately 470 nm placed at a distance of approximately 140 mm under the fluid sample.
5. The method of claim 1 , further comprising the specific light-dark pattern across the plurality of blurry images comprising, in forward or reverse order:
(a) a dark ring around a bright center;
(b) a dark circular shape; and
(c) a white center with dark ring.
6. The method of claim 1 , the specific light-dark pattern across the plurality of blurry images comprising, in forward or reverse order:
(a) a black ring surrounding a white core; and
(b) the white core growing smaller and turning black, with a white ring surrounding it.
7. The method of claim 1 , the specific light-dark pattern across the plurality of blurry images comprising, in forward or reverse order:
(a) a black ring surrounding a greyish core; and
(b) the greyish core growing smaller and turning black, with a white ring surrounding it.
8. The method of claim 1 , the specific light-dark pattern across the plurality of blurry images comprising, in forward or reverse order:
(a) a grey center and an outside ring of a darker grey;
(b) a smaller white center surrounded by a dark grey or black ring; and
(c) a dark grey center surrounded by a large grey outside ring.
9. The method of claim 1 , the specific light-dark pattern across the plurality of blurry images comprising, in forward or reverse order:
(a) a completely grey circle;
(b) a black circle; and
(c) a white core surrounded by a grey circle.
10. The method of claim 1 , wherein the organic particles are platelets.
11. The method of claim 1 , wherein the organic particles are blood cells.