Systems and methods for high-throughput radiation biodosimetry

• US 7,822,249 B2
• Filed: 08/24/2007
• Issued: 10/26/2010
• Est. Priority Date: 08/25/2006
• Status: Expired due to Fees

• Current Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Original Assignee: Trustees Of Columbia University In The City Of New York (Columbia University)
• Inventors: Bhatla, Anubha, Lyulko, Oleksandra V., Simaan, Nabil, Brenner, David J., Randers-Pehrson, Gerhard, Zhang, Jian, Garty, Guy, Yao, Y. Lawrence, Salerno, Alessio, Dutta, Aparajita
• Primary Examiner(s): SHERALI, ISHRAT I
• Application Number:
  US11/895,417
• Publication Number:
  US 20080181473A1
• Time in Patent Office: 1,159 Days
• Field of Search: 382128-134, 382/255, 382/153, 422 63- 43, 700245-249, 378 62- 67, 600/300, 600/3, 600/407, 600/584, 604/4.1, 604/141, 604/151, 604/19, 506/7, 324/316, 705/2, 702 19- 22, 435/69.1, 435/288.7, 21912167-12169
• US Class Current: 382/128
• CPC Class Codes: C40B 30/10 by measuring physical properties, e.g. mass
• CPC Class Codes: C40B 60/12 for screening libraries

Abstract

Systems and methods for high-throughput radiation biodosimetry are disclosed herein. In some embodiments, a high-throughput methods of analyzing a population for radiation exposure can include, in various possible sequences: marking a first capillary designed to retain a first sample from the population with a first identifier; transporting a plurality of samples to a biodosimetry system; inputting the samples into the biodosimetry system; centrifuging the plurality of samples including the first sample wherein each sample can be retained in a capillary and the first sample can be retained in the first capillary; transferring the plurality of capillaries including the first capillary from the centrifuge to a cutting device using a robotic device; cutting the first capillary; reading the first identifier; transferring at least one portion of the first sample from the first capillary to a well in an array, wherein the array can include one or more filters in a multi-well plate; correlating the first identifier to a location of the array that includes the at least one portion of the first sample; one or more cycles of biological processing, which can include addition of a reagent and/or incubation of a selected temperature such as, for example, 37° C., 4° C., room temperature, and the like; sealing the array; positioning the array adjacent to an imaging element; focusing the imaging element; capturing an image of the first sample in the array; and analyzing the image to determine whether the first sample indicates a level of radiation exposure exceeding a predetermined threshold.

Claims

1. A high-throughput method of analyzing a plurality of biological samples, comprising:

(a) marking an identifier onto each of a plurality of capillary vessels;

(b) collecting a biological sample in each of the plurality of capillary vessels;

(c) transporting a receptacle to a centrifuge using a first robotic device, said receptacle containing the plurality of capillary vessels;

(d) extracting a predetermined component from each biological sample;

(e) image scanning the plurality of biological samples, said image scanning comprising detecting an image of each sample with an optical device;

(f) focusing the image with the optical device; and

(g) determining a radiation exposure of an organism from which the sample originated by analyzing the focused image.

4. The method of claim 1, wherein said extracting comprises:

centrifuging the receptacle;

transferring the receptacle from the centrifuge to a cutting device using a second robotic device;

reading the identifier marking on each of the plurality of capillary vessels;

cutting each of the plurality of capillary vessels using the cutting device;

transferring at least a portion of each of the plurality of biological samples from each of the capillary vessels to a corresponding well in a multi-well plate using a third robotic device;

correlating each identifier marking to a corresponding well; and

performing a biological process on each of the plurality of biological samples.

5. The method of claim 4, wherein said cutting comprises focusing a laser on a predetermined point on the capillary vessel and cutting the capillary vessel using a laser at the predetermined point.

6. The method of claim 4, wherein said multi-well plate includes a plurality of filter-bottomed wells arranged in an array.

7. The method of claim 4, wherein said biological process comprises at least one of adding a reagent to each said sample and incubating said sample.

8. The method of claim 1, wherein said image scanning comprises:

detecting an image of a sample in the multi-well plate with an optical device by directing the image toward a sensor;

controlling the position of a first mirror and the position of a second mirror using a processor; and

detecting the image of the sample with the optical device using the sensor positioned relative to the first mirror and the second mirror.

9. The method of claim 8, wherein said directing comprises:

positioning the first mirror relative to the sample, wherein the first mirror directs a portion of the image of the sample in a first direction; and

positioning the second mirror relative to the sample and the first mirror, wherein the second mirror directs the image of the sample in a second direction.

10. The method of claim 1, wherein said focusing comprises:

collecting light from a region of the sample with an objective lens, said region having a feature with a known geometric characteristic;

splitting the collected light into a first portion and a second portion, and directing said first portion through a weak cylindrical lens to a focusing sensor, and directing said second portion to an imager;

observing, with said focusing sensor, a shape of the feature;

focusing the optical device by moving at least one of the objective lens and the object to be imaged until the observed shape of the feature has a predetermined relationship to the known geometric characteristic; and

acquiring a focused image of the sample.

18. The method of claim 1, wherein the identifier is marked onto each of the plurality of capillary vessels by etching.

2. An apparatus for high-throughput analysis of a plurality of biological samples, comprising:

(a) means for marking an identifier onto each of a plurality of capillary vessels;

(b) a plurality of capillary vessels, each containing a single one of the plurality of biological samples;

(c) a robotic device for transporting a receptacle to a centrifuge, said receptacle containing the plurality of capillary vessels;

(d) an extraction apparatus for extracting a predetermined component from each biological sample;

(e) an image scanning apparatus for detecting an image of the extracted component;

(f) a focusing apparatus for focusing the image with an optical device; and

(g) a processor for determining a radiation exposure of an organism from which the sample originated by analyzing the focused image.

11. The apparatus of claim 2, wherein said extraction apparatus comprises:

a centrifuge for centrifuging the receptacle;

a second robotic device for transferring the receptacle from the centrifuge to a cutting device;

means for reading the identifier marking on each of the plurality of capillary vessels;

a third robotic device for transferring at least a portion of each of the plurality of biological samples from each of the capillary vessels to a corresponding well in a multi-well plate;

means for correlating each identifier marking to a corresponding well; and

means for performing a biological process on the biological samples.

12. The apparatus of claim 11, wherein said cutting device comprises:

a laser; and

a focusing means for focusing the laser on a predetermined point of the capillary.

13. The apparatus of claim 11, wherein said multi-well plate includes a plurality of filter-bottomed wells arranged in an array.

14. The apparatus of claim 11, wherein said biological process comprises at least one of adding a reagent to each said sample and incubating each said sample.

15. The apparatus of claim 2, wherein the image scanning apparatus comprises:

means for directing the image toward a sensor, and an optical device; and

a processor for controlling the position of a first mirror and the position of a second mirror, and wherein the sensor is positioned relative to the first mirror and the second mirror.

16. The apparatus of claim 15, wherein the first mirror is positioned relative to the sample for directing a portion of the image of the sample in a first direction relative to the sample, and wherein the second mirror is positioned relative to the sample and the first mirror, for directing the image of the sample in a second direction.

17. The apparatus of claim 2, wherein said focusing apparatus comprises:

an objective lens for collecting light from a region of the sample to be imaged, said region having a feature with a known geometric characteristic;

means for splitting the collected light into a first portion and a second portion, and directing said first portion through a weak cylindrical lens to a focusing sensor for observing a shape of the feature, and directing said second portion to an imager;

means for focusing the optical device by moving at least one of the objective lens and the object to be imaged until the observed shape of the feature has a predetermined relationship to the known geometric characteristic; and

means to acquire a focused image of the sample.

19. The apparatus of claim 2, wherein the means for marking an identifier marks the capillary vessels by etching.

3. A system for high-throughput analysis of a plurality of biological samples, comprising:

(a) means for marking an identifier onto each of a plurality of capillary vessels, each capillary vessel containing a single one of the plurality of biological samples;

(b) means for transporting a receptacle to a centrifuge, said receptacle containing the plurality of capillary vessels;

(c) means for extracting a predetermined element from each biological sample, each said extracted element being correlated to a respective source of the biological sample;

(d) means for detecting an image of each biological sample;

(e) means for focusing the image; and

(g) means for analyzing the focused image to determine a radiation exposure of an organism, said organism being the source of the biological sample.

20. The system of claim 3, wherein the means for marking an identifier marks the capillary vessels by etching.

21. The system of claim 3, wherein each of the plurality of capillary vessels contains a biological sample from a population, and further comprising:

(a) a receptacle, said receptacle containing the plurality of capillary vessels;

(b) a centrifuge;

(c) a first robotic device for transporting the receptacle between an input module and the centrifuge;

(d) a second robotic device for transporting the receptacle between the centrifuge and a sample harvest location;

(e) a cutting device for cutting each of the plurality of capillary vessels;

(f) a multi-well plate having a plurality of wells arranged in an array; and

(g) a third robotic device for transferring at least one portion of each of the plurality of biological samples from each of the plurality of capillary vessels to a corresponding well in the array.

22. The system of claim 3, wherein each of the plurality of capillary vessels contains a biological sample from a population, and further comprising:

(a) a holding means for holding the plurality of capillary vessels;

(b) a centrifuge means for separating each of the biological samples into a plurality of elements;

(c) a first transporting means for transporting the holding means, including the plurality of capillary vessels, to the centrifuge means;

(d) a second transporting means for transporting the receptacle from the centrifuge to a cutting location;

(e) a cutting means for cutting each of the plurality of capillary vessels at the cutting location;

(f) a holding means having a plurality of locations, each of the plurality of locations for holding at least one portion of one of the plurality of biological samples; and

(g) a transferring means for transferring at least one portion of each of the plurality of biological samples from each of the plurality of capillary vessels to a corresponding location in the holding means.