Method and apparatus for optical analysis
An optical analysis apparatus, including: a sample delivery system from which a liquid sample may be delivered in operation; a flow cell defining a channel through which, in operation, the delivered liquid sample may flow at a controllable rate, the channel including an optical analysis region; an illumination source focused on a portion of the optical analysis region that, in operation, illuminates a single particle at a time in a stream of the sample wider than the single particle; a detector that, in operation, detects light resulting from the illumination of the sample and outputting a signal representative of the detected light; and an analysis system receiving the representative signal.
1. An optical analysis apparatus, comprising:
(a) a sample delivery system from which a liquid sample may be delivered;
(b) a flow cell defining a channel including an optical analysis region and through which the liquid sample flows;
(c) an illumination source;
(d) a controllable moveable illumination scanning mechanism wherein:
(i) the structured movement traverses and illuminates the optical analysis region in a plurality of directions wherein the light originates from the illumination source;
(ii) the movement is structured to make a measurement of a particle and generate an image of the particle;
(e) an objective lens that conveys light from the illumination source to the liquid sample, images light emanating from the illuminated optical analysis region of the liquid sample onto an image plane, and through which light emanating from the sample in response to the illumination is detected;
(f) a detector to detect light emanating from the sample and collected by the objective lens in response to the illumination of the sample; and
(g) an aperture placed in an image plane of the objective lens to spatially filter light from points other than the focal plane of the objective lens, which is within the liquid sample flow, before the light reaches the detector.
2. The optical analysis apparatus of claim 1 wherein the movement of the illumination traversing the optical analysis region includes an X vector perpendicular to a direction of the liquid sample flow and a Y vector parallel to the direction of the liquid sample flow and further that a velocity of movement of the Y vector may be substantially equal to a velocity of the liquid sample flow.
3. The optical analysis apparatus of claim 2 wherein the two-dimensional pattern of the traversing optical scanner achieves consistency of illumination.
4. The optical analysis apparatus of claim 1 wherein the extent of the traverse in the X or Y direction or rate of traverse of the optical analysis region can be controlled in real time.
5. The optical analysis apparatus of claim 2 wherein the traversable optical scanner utilizes a lemniscate pattern.
6. The optical analysis apparatus of claim 5 wherein the Y location of the optical scanner is reset after each traverse in the X direction.
7. The optical analysis apparatus of claim 1 wherein the two-dimensional traversing optical scanner achieves multiple two-dimensional images of the sample.
8. The optical analysis apparatus of claim 1 wherein the sample may be illuminated by two illumination sources wherein light from the illumination sources is reflected first by a first dichroic beamsplitter, by a second dichroic beamsplitter, and then by a scanning mirror attached to the optical scanner.
9. The optical analysis apparatus of claim 8 further comprising a scanning mirror rotating on an axis orthogonal to the sample flow direction.
10. The optical analysis apparatus of claim 8 further comprising a second scanning mirror thereby allowing the apparatus to scan in two dimensions.
11. The optical scanner of claim 8 wherein the optical scanner may further comprise a scan lens, f-theta lens or a field lens.
12. An optical analysis apparatus comprising:
(a) a sample delivery system comprising:
(i) a delivery component for delivery of a liquid sample;
(ii) a flow cell component defining a channel to which the liquid sample is delivered;
(b) combination of an illumination source, moveable scanning mechanism, and an objective lens wherein the illumination source, moveable scanning mechanism and objective lens are structured to illuminate an optical analysis region of the flow cell to make a measurement of a particle and generate an image of the particle; and
(c) the controllable moveable scanning mechanism further traverses the optical analysis region in a plurality of directions.
13. The optical analysis apparatus of claim 12 wherein the flow control component can control the sample flow rate through the optical analysis region in real time.
14. The optical analysis apparatus of claim 12 wherein the flow control component can control the sample flow rate through the optical analysis region in real time.
15. A method of conducting optical analysis, comprising:
(a) delivering a sample into a flow cell defining a channel including an optical analysis region and through which the liquid sample flows;
(b) illuminating the liquid sample with an illumination source and scanning mechanism;
(c) controlling the moveable illumination scanning mechanism wherein the moveable illumination scanning mechanism traverses the optical analysis region in a plurality of directions;
(d) creating an image using an objective lens that conveys light from the illumination source to the liquid sample, imaging the light emanating from the illuminated optical analysis region of the liquid sample onto an image plane and through which light emanating from the sample in response to the illumination is detected; and
(e) utilizing an aperture placed in an image plane of the objective lens to spatially filter light from points other than the focal plane of the objective lens, which is within the liquid sample flow, before the light reaches the detector.
16. The method of claim 15 further comprising moving the illumination scanning mechanism to traverse the optical analysis region in an X vector perpendicular to a direction of the liquid sample flow and a Y vector parallel to the direction of the liquid sample flow.
17. The method of claim 16 including moving the illumination scanning mechanism in the Y vector at a velocity substantially equal to a velocity of the liquid sample flow.
18. The method of claim 15 further comprising controlling in real time the extent of the traverse in the X or Y direction or rate of traverse of the optical analysis region.
19. The method of claim 15 further comprising utilizing a lemniscate pattern to traverse the optical analysis region.
20. The method of claim 19 further comprising resetting the Y location of the optical scanner apparatus after each traverse in the X direction.
21. A method of conducting optical analysis comprising:
(a) a sample delivery system comprising:
(i) a delivery component for delivery of a liquid sample;
(ii) a flow cell component defining a channel to which the liquid sample is delivered;
(b) illuminating the sample in an optical analysis region of the flow cell;
(c) controlling a controllable moveable scanning mechanism in a plurality of directions to substantially synchronize with the velocity of flow; and
(d) imaging at least a portion of the illuminated sample within the optical analysis region with an objective lens.