Systems and methods for high-throughput radiation biodosimetry

• US 20080181473A1
• Filed: 08/24/2007
• Published: 07/31/2008
• Est. Priority Date: 08/25/2006
• Status: Active Grant

• 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: Simaan, Nabil, Brenner, David J., Randers-Pehrson, Gerhard, Zhang, Jian, Garty, Guy, Yao, Y. Lawrence, Salerno, Alessio, Bhatla, Anubha, Lyulko, Oleksandra V., Dutta, Aparajita
• Granted Patent: US 7,822,249 B2
• Time in Patent Office: Days
• 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) etching an identifier marking 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, 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, wherein said cutting step comprises focusing a laser on a predetermined point on the capillary vessel and cutting the capillary vessel using a laser at the predetermined point;

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, said multi-well plate having a plurality of filter-bottomed wells arranged in an array;

correlating each identifier marking to a corresponding well;

performing a biological process on each of the plurality of biological samples, said biological process comprising at least one of adding a reagent to each said sample and incubating said sample;

(e) image scanning the plurality of biological samples, said image scanning comprising;

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

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

controlling the position of the first mirror and the position of the 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;

(f) focusing the image with the optical device, said focusing comprising;

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;

acquiring a focused image of the sample;

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

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

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

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

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

(d) extraction apparatus for extracting a predetermined component from each biological sample, said extraction apparatus comprising;

a centrifuge for centrifuging the receptacle;

a second robotic device for transferring the receptacle from the centrifuge to a cutting device, said cutting device comprising a laser and focusing means for focusing the laser on a predetermined point on the capillary;

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, said multi-well plate having a plurality of filter-bottomed wells arranged in an array;

means for correlating each identifier marking to a corresponding well;

means for performing a biological process on the biological samples, said biological process comprising at least one of adding a reagent to each said sample and incubating each said sample;

(e) image scanning apparatus for detecting an image of the extracted element in the multi-well plate, comprising means for directing the image toward a sensor, and an optical device, said image scanning apparatus further comprising;

a first mirror positioned relative to the sample for directing a portion of the image of the sample in a first direction relative to the sample, and

a second mirror positioned relative to the sample and the first mirror, for directing the image of the sample in a second direction;

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

wherein the sensor is positioned relative to the first mirror and the second mirror;

(f) focusing apparatus for focusing the image with the optical device, said focusing apparatus comprising;

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;

means to acquire a focused image of the sample;

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

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

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

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

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

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

(f) means for focusing the image;

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

4. A method of determining radiation exposure of an organism comprising:

(a) collecting a biological sample from the organism using a minimally invasive technique;

(b) extracting a predetermined element from the sample;

(c) capturing an image of the predetermined element; and

(d) analyzing the image to determine radiation exposure of the organism.

5. A system for determining radiation exposure of an organism comprising:

(a) a biological sample from the organism;

(b) a centrifuge adapted to separate the different elements contained in the biological sample

(c) a liquid handling and incubation device capable of adding reagents;

(d) an imaging element adapted to capture an image of a predetermined element from the sample; and

(e) a processor adapted to analyze the image to determine radiation exposure of the sample.

6. A method of marking a cylindrical capillary comprising:

(a) etching a marking on to the capillary using a first laser; and

(b) reading the marking using a second laser.

7. An apparatus for identifying a capillary vessel comprising:

(a) a first laser adapted to mark the capillary vessel; and

(b) a second laser adapted to read the mark.

8. A system for identifying a capillary tube comprising:

(a) a means for marking the capillary tube by etching; and

(b) a means for reading the etched marking.

9. A method of marking a cylindrical capillary comprising:

(a) etching a marking on to the capillary using a laser; and

(b) reading the marking using a charge-coupled device (CCD).

10. A method of marking a cylindrical capillary comprising:

(a) etching a marking on to the capillary using a laser; and

(b) reading the marking using a complementary metal-oxide-semiconductor device (CMOS).

11. A robotic system for transporting biological samples, comprising:

(a) a plurality of capillary vessels, each capillary vessel containing a biological sample from a population;

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

(c) a centrifuge;

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

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

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

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

(h) 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.

12. A method for transporting biological samples using a robotic system, comprising:

(a) transporting a receptacle to a centrifuge using a first robotic device, said receptacle containing a plurality of capillary vessels and each said capillary vessel containing a biological sample;

(b) centrifuging the receptacle;

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

(d) cutting each of the plurality of capillary vessels using the cutting device; and

(e) 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, said multi-well plate having a plurality of wells arranged in an array.

13. A robotic system for transporting biological samples, comprising:

(a) a plurality of capillary vessels, each capillary vessel containing a biological sample from a population;

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

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

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

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

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

(g) 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

(h) 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.

14. A method of at least partially severing a capillary vessel comprising:

(a) focusing a laser on a predetermined point on the capillary vessel, said capillary vessel containing a biological sample; and

(b) cutting the capillary vessel using a laser at the predetermined point.

15. An apparatus for at least partially bisecting a capillary comprising:

(a) an imaging element adapted to capture an image of the capillary;

(b) a processor adapted to analyze the image and determine a cutting point based on the image; and

(c) a laser adapted to cut the capillary at the cutting point.

16. A system for at least partially severing a capillary vessel comprising:

(a) an imaging means for capturing an image of the capillary vessel;

(b) a processing means for determining a cutting point based on the image; and

(c) a cutting means for cutting the capillary at the cutting point.

17. A method for focusing an optical device, comprising:

(a) collecting light from a region of an object to be imaged with an objective lens, said region having a feature with a known geometric characteristic;

(b) 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;

(c) observing, with said focusing sensor, a shape of the feature; and

(d) 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.

18. An apparatus for focusing an optical device, comprising:

(a) an objective lens for collecting light from a region of an object to be imaged through an objective lens, said region having a feature with a known geometric characteristic;

(b) 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, and directing said second portion to an imager;

(c) a focusing sensor for observing a shape of the feature;

(d) a mechanism for focusing the optical device by moving at least one of the objective lens and the object to be imaged; and

(e) a processor for analyzing the observed shape and determining whether the observed shape of the feature has a predetermined relationship to the known geometric characteristic.

19. A system for focusing an optical device, comprising:

(a) a light collecting means for collecting light from a region of an object to be imaged with an objective lens, said region having a feature with a known geometric characteristic;

(b) a light splitting 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, and directing said second portion to an imager;

(c) mechanical means for focusing the optical 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

(d) a processing means, coupled to the mechanical means and focusing sensor, for analyzing the observed shape and determining whether the observed shape of the feature has a predetermined relationship to the known geometric

20. A method for high speed image scanning, comprising:

(a) adjusting an object using a positioning element;

(b) directing a portion of an image of the object toward a sensor by positioning a first mirror relative to the object, wherein the first mirror directs the portion of the image of the object in a first direction, and by positioning a second mirror relative to the object and the first mirror, wherein the second mirror directs the portion of the image of the object in a second direction;

(c) controlling the positioning element, the position of the first mirror and the position of the second mirror using a processor; and

(d) detecting the portion of the image of the object using the sensor positioned relative to the first mirror and the second mirror.

21. An apparatus for high speed image scanning, comprising:

(a) an imaging element;

(b) a positioning element controllable in an x direction and in a y direction, coupled to an object, wherein the positioning element adjusts the position of the object;

(c) a first mirror positioned relative to the object;

(d) a second mirror positioned relative to the object and the first mirror, wherein the first mirror directs at least one image of the object in the x direction toward the second mirror and wherein the second mirror directs the at least one image of the object in the y direction toward a sensor; and

(e) a processor coupled to the positioning element, the first mirror and the second mirror, wherein the processor controls the positioning element, the position of the first mirror and the position of the second mirror.

22. A system for high speed image scanning, comprising:

(a) an imaging means for providing a magnified image of an object;

(b) a positioning means controllable in an x direction and in a y direction, coupled to an object, wherein the positioning means adjusts the position of the object;

(c) a first reflecting means positioned relative to the object;

(d) a second reflecting means positioned relative to the object and the first reflecting means, wherein the first reflecting means directs at least one image in the x direction toward the second reflecting means and wherein the second reflecting means directs the at least one image of the object in the y direction toward a sensing means; and

(e) a processing means coupled to the positioning means, the first reflecting means and the second reflecting means, wherein the processing means controls the positioning means, the position of the first reflecting means and the position of the second reflecting means.