Methods for analyzing inter-cellular phenomena
View Patent ↗Aspects of the present invention encompass the collection of multispectral images from a population of objects, and the analysis of the collected images to measure at least one characteristic of the population, using photometric and/or morphometric features identifiable in the collection of images. In an exemplary application, the objects are biological cells. In a particularly preferred, but not limiting implementation, the plurality of images for each individual object are collected simultaneously. In an empirical study, the characteristic being measured involves the synapse between conjugated cells. The conjugated cells may represent a subpopulation of the overall population of objects that were imaged. In a particularly preferred, yet not limiting embodiment, the present invention enables the quantization of the redistribution of cellular molecules due to the conjugation of different biological cells. Significantly, such quantization is not feasible with standard microscopy and flow cytometry.
1. A method for determining one or more characteristics of a subpopulation of objects from images collected from a population of objects, comprising the steps of:
(a) imaging the population of objects to collect image data, while there is relative movement between the population of objects and an imaging system used to image the population of objects, where each object in the population of objects is imaged using the following steps:
(i) dispersing light from the object into a plurality of light beams;
(ii) focusing the plurality of light beams to produce respective images, each respective image corresponding to a different one of the plurality of light beams; and
(iii) providing a time delay integration (TDI) detector disposed to receive the respective images, the steps of dispersing the light and providing the TDI detector being implemented such that the dispersed light is dispersed across the detector in a direction that is substantially orthogonal to a direction of a motion of the respective images across the TDI detector, enabling a plurality of different images of the object to be acquired simultaneously;
(b) using the image data to define the subpopulation of objects, the subpopulation of objects being encompassed by the population of objects;
(c) identifying an object feature exhibited by the subpopulation of objects; and
(d) using the image data to measure at least one characteristic associated with the object feature.
2. The method of claim 1 , wherein the step of simultaneously collecting a plurality of images for each object in the population of objects comprises the step of simultaneously collecting at least two of the following types of images: a brightfield image, a darkfield image, and a fluorescent image.
3. The method of claim 1 , wherein the population of objects comprises at least two different types of objects.
4. The method of claim 3 , wherein before the population of objects is imaged, further comprising the step of labeling at least one of the at least two different types of objects.
5. The method of claim 3 , wherein the objects comprise biological cells.
6. The method of claim 3 , wherein the population of objects comprise T cells and antigen presenting cells.
7. The method of claim 3 , wherein the step of using the image data to define the subpopulation of objects comprises a step of defining the subpopulation to comprise conjugated cells.
8. The method of claim 7 , wherein the step of identifying the object feature exhibited by the subpopulation of objects comprises the step of identifying a synapse between the conjugated cells.
9. A method for determining one or more characteristics of a subpopulation of objects from images collected from a population of objects, comprising the steps of:
(a) imaging the population of objects to collect image data, while there is relative movement between the population of objects and an imaging system used to image the population of objects, such that a plurality of images for individual objects in the population of objects are simultaneously collected, the plurality of images comprising at least two of the following types of images: a brightfield image, a darkfield image, and a fluorescent image;
(b) using the image data to define the subpopulation of objects, the subpopulation of objects being encompassed by the population of objects;
(c) identifying an object feature exhibited by the subpopulation of objects; and
(d) using the image data to measure at least one characteristic associated with the object feature.
10. The method of claim 9 , wherein before the population of objects is imaged, further comprising the step of labeling a plurality of individual objects encompassed by the population of objects.
11. The method of claim 9 , wherein the population of objects comprises at least two different types of objects.
12. The method of claim 11 , wherein before the population of objects is imaged, further comprising the step of labeling at least one of the at least two different types of objects.
13. The method of claim 11 , wherein the population of objects comprise T cells and antigen presenting cells.
14. The method of claim 11 , wherein the step of using the image data to define the subpopulation of objects comprises a step of defining the subpopulation to comprise conjugated cells.
15. The method of claim 11 , wherein the step of identifying the object feature exhibited by the subpopulation of objects comprises the step of identifying a synapse between the conjugated cells.
16. The method of claim 9 , wherein each individual object in the population of objects is imaged using the following steps:
(a) dispersing light from the object into a plurality of light beams;
(b) focusing the plurality of light beams to produce respective images, each different respective image corresponding to one of the plurality of light beams; and
(c) providing a time delay integration (TDI) detector disposed to receive the respective images, the steps of dispersing the light and providing the TDI detector being implemented such that the dispersed light is dispersed across the detector in a direction that is substantially orthogonal to a direction of a motion of the respective images across the TDI detector.
17. A method for determining one or more characteristics shared by a group of objects, comprising the steps of:
(a) imaging a population of objects to collect image data, while there is relative movement between the population of objects and an imaging system used to image the population of objects, such that a plurality of images for individual objects in the population of objects are simultaneously collected, such that the plurality of images for an individual object are not optically combined when they are collected;
(b) identifying an object feature exhibited by a group of objects encompassed in the population of objects; and
(c) using the image data to measure at least one characteristic associated with the object feature.
18. The method of claim 17 , wherein each individual object in the population of objects is imaged using the following steps:
(a) dispersing light from the object into a plurality of light beams;
(b) focusing the plurality of light beams to produce respective images, each respective image corresponding to a different one of the plurality of light beams; and
(c) providing a time delay integration (TDI) detector disposed to receive the respective images, the steps of dispersing the light and providing the TDI detector being implemented such that the dispersed light is dispersed across the detector in a direction that is substantially orthogonal to a direction of a motion of the respective images across the TDI detector.
19. A method for determining one or more characteristics of a subpopulation of objects from images collected from a population of objects, comprising the steps of:
(a) imaging the population of objects to collect image data, while there is relative movement between the population of objects and an imaging system used to image the population of objects, such that for each object, a plurality of individual images are simultaneously collected;
(b) using the image data to define the subpopulation of objects, the subpopulation of objects being encompassed by the population of objects;
(c) identifying an object feature exhibited by the subpopulation of objects; and
(d) using the image data to measure at least one characteristic associated with the object feature.