SYSTEM AND METHOD FOR AUTOMATED DIAGNOSIS
An automated microscope apparatus comprises an outer housing having an external wall; optionally but preferably an internal wall in the housing configured to form a first compartment and a separate second compartment in the outer housing; a microscope assembly in the housing (preferably in the first compartment); a microprocessor in the housing (preferably in the second compartment), and (optionally but preferably) a heat sink mounted on the housing external wall, preferably adjacent the second compartment, with the microprocessor thermally coupled to said heat sink and operatively associated with the microscope assembly. Systems and methods employing the same are also described, along with component parts thereof.
1 . An automated method to aid detecting a disorder in a subject, comprising:
securing a sample cartridge to the stage of an automated microscope; said sample cartridge comprising at least one chamber, said at least one chamber containing a biological sample collected from a subject, and optionally exogeneous targets;
autofocusing said microscope on said at least one chamber;
imaging selected cells in said sample, said selected cells including at least neutrophils;
generating a count of at least neutrophils in said sample as an aid to detecting a disorder in said subject; and
optionally repeating at least said imaging step for at least one additional chamber on said cartridge.
2 . The method of claim 1 , wherein said disorder is an infection or inflammation.
3 . The method of claim 1 , wherein said sample cartridge comprises a plurality of chambers and said method further comprises the steps of:
inputting selection of which of said chambers are to be imaged prior to said autofocusing step; and
repeating said imaging step for said at least one additional chamber based on said input selection.
4 . The method of claim 1 , wherein said cartridge comprises two chambers.
5 . The method of claim 1 , wherein said cartridge comprises four chambers.
6 . The method of claim 1 , wherein said step of generating a count comprises generating a separate count for (a) neutrophils, (b) lymphocytes, and (c) macrophages.
7 . The method of claim 1 , further comprising the step of:
determining the presence or absence of a disorder in said subject from said cell count.
8 . The method of claim 7 , wherein said determining step is carried out by differential leukocyte analysis.
9 . The method of claim 1 , wherein:
said subject is a dairy animal,
said disorder is mastitis,
said biological sample is selected from the group consisting of milk, secretions, or colostrum,
said sample cartridge comprises from two to four chambers, and/or
each chamber contains a milk sample collected from a different teat of said dairy animal.
10 . The method of claim 1 , wherein said cartridge comprises a plurality of chambers, and said autofocusing step is carried out separately for each of said chambers.
11 . The method of claim 1 , wherein said autofocusing step comprises:
capturing an image from each of a plurality of focal planes in or on said specimen, with a plurality of exogeneous targets in or on said specimen;
calculating a focus score for each of said images,
selecting the focal plane corresponding to the image having the best focus score, and then
repositioning said specimen relative to said microscope so that said microscope is focused on said selected focal plane.
12 . The method of claim 11 , wherein said exogenous targets are particles and have an average diameter of from 0.1 micrometers up to 10 micrometers.
13 . The method of claim 11 , wherein said exogenous targets are fluorescent.
14 . The method of claim 13 , wherein said exogenous targets:
fluoresce at a peak absorption wavelength of at least 420 nanometers and at not more than 540 nanometers;
fluoresce at a peak emission wavelength of at least 450 nanometers not more than 590 nanometers; and
and wherein said peak absorption wavelength and said peak emission wavelength differ by at least 10 nanometers.
15 . An automated system for detecting a disorder in a subject, comprising:
an XYZ stage configured to secure a sample cartridge; said sample cartridge comprising at least one chamber, said at least one chamber containing a biological sample collected from a subject;
an imaging system operatively associated with said XYZ stage and configured to image selected cells in said sample, said selected cells including at least neutrophils;
an autofocusing system operatively associated with said imaging system and said XYZ stage and configured to focus said imaging system on said at least one chamber;
means for generating a count of at least neutrophils in said sample as an aid to detecting a disorder in said subject; and
a controller configured to optionally repeat at least said imaging for at least one additional chamber on said cartridge.
16 . The system of claim 15 , wherein said disorder is an infection or inflammation.
17 . The system of claim 15 , wherein said sample cartridge comprises a plurality of chambers and said system further comprises:
a user interface for inputting selection of which of said chambers are to be imaged prior to said autofocusing step; and
a controller operatively associated with said user interface and configured for repeating said imaging step for said at least one additional chamber based on said input selection.
18 . The system of claim 15 , wherein said means for generating a count comprises means for generating a separate count for (a) neutrophils, (b) lymphocytes, and (c) macrophages.
19 . The system of claim 15 , further comprising:
means for determining the presence or absence of a disorder in said subject from said cell count.
20 . The system of claim 19 , wherein said determining is carried out by differential leukocyte analysis.
21 . The system of claim 20 , wherein:
said subject is a dairy animal,
said disorder is mastitis,
said biological sample is selected from the group consisting of milk, secretions, or colostrum,
said sample cartridge comprises from two to four chambers, and/or
each chamber contains a milk sample collected from a different teat of said dairy animal.
22 . The system of claim 15 , wherein the cartridge contains exogeneous targets, and wherein said means for autofocusing is responsive to said exogenous targets.
23 . The system of claim 22 , wherein said exogenous targets are particles and have an average diameter of from 0.1 micrometers up to 10 micrometers.
24 . The system of claim 23 , wherein said exogenous targets are fluorescent.
25 . The system of claim 24 , wherein said exogenous targets:
fluoresce at a peak absorption wavelength of at least 420 nanometers and at not more than 540 nanometers;
fluoresce at a peak emission wavelength of at least 450 nanometers not more than 590 nanometers; and
and wherein said peak absorption wavelength and said peak emission wavelength differ by at least 10 nanometers.