System for imaging captured cells

• US 9,612,199 B2
• Filed: 06/30/2016
• Issued: 04/04/2017
• Est. Priority Date: 03/13/2013
• Status: Active Grant

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Denovo Sciences, Inc. (Bio-Rad Laboratories Incorporated)
• Inventors: Handique, Kalyan, Gleason, Kyle, Siemer, Christopher, Gogoi, Priyadarshini, Javdani, Saedeh Sepehri, Parunak, Gene, Kehrer, Aaron, Meines, Jon
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): FISHER, BRITTANY I
• Application Number:
  US15/199,245
• Publication Number:
  US 20160341666A1
• Time in Patent Office: 278 Days
• CPC Class Codes: A61B 5/00 Measuring for diagnostic purposes radiation diagnosis A61B6/00; diagnosis by ultrasonic, sonic or infrasonic waves A61B8/00; Identification of persons
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 33/4833 of solid biological material, e.g. tissue samples, cell cultures tissue in vivo A61B5/00; cell suspensions G01N33/48735
• CPC Class Codes: G01N 33/48735 Investigating suspensions of cells, e.g. measuring microbe concentration by chemical means C12Q1/04; colony counters C12M1/34; concentration of particle suspensions in general G01N15/06
• CPC Class Codes: G02B 21/06 Means for illuminating specimens
• CPC Class Codes: G02B 21/26 Stages; Adjusting means therefor

Abstract

A system for imaging captured cells comprising: an illumination module configured to illuminate a target object; a platform configured to position the target object in relation to the illumination module; a filter module configured to filter light transmitted to the target object and/or to filter light received from the target object, an optical sensor configured to receive light from the target object and to generate image data; and a focusing and optics module configured to manipulate light transmitted to the optical sensor. The system can further comprise one or more of: a control system configured to control at least one of the illumination module, the platform, the focusing and optics module, the filter module, and the optical sensor; a tag identifying system configured to identify and communicate tag information from system elements; a thermal control module configured to adjust temperature parameters of the system; and an image stabilization module.

Claims

1. A system for detecting biological sample features comprising:

a platform including a set of guides that receive a set of imaging substrates with a set of biological samples;

a filter module superior to the platform including an excitation filter, an emission filter, and a dichroic mirror;

a first light source, inferior to the platform, configured to transmit light toward a biological sample of the set of biological samples from a first direction;

a second light source configured to transmit light through the excitation filter, to be reflected from the dichroic mirror, and to reach the biological sample from a second direction opposed to the first direction;

a lens, coupled to a lens selector, that focuses light from the excitation filter onto a target object of the biological sample, and transmits light from the target object, through the dichroic mirror and the emission filter, toward an optical sensor;

a platform control module configured to rotate the platform about a first axis perpendicular to a broad surface of the platform and translate the platform along a direction perpendicular to the first axis, into a first configuration that aligns at least one imaging substrate between the first light source and the lens;

a filter stage, situated superior to the platform, configured to rotate the filter module about a second axis, parallel to the first axis, into a second configuration that positions the filter module to receive light from the second light source and to transmit light toward the optical sensor; and

an optics manipulation module, including a translation stage coupled to an optical shaft concentrically aligned with the lens by way of the lens selector, configured to vertically translate the lens to facilitate focusing of the target object.

2. The system of claim 1, wherein the set of guides is distributed about the platform in a first circular array, wherein the system further includes a set of filter modules distributed about the filter stage in a second circular array, wherein the platform control module is configured to rotate the platform into a first set of configurations that align each guide of the set of guides between the first light source and the lens, and wherein the filter stage is configured to rotate the set of filter modules into a second set of configurations that align each filter module of the set of filter modules to receive light from the second light source and transmit light toward the optical sensor.

3. The system of claim 2, wherein the platform control module includes an actuator configured to translate the platform along two directions perpendicular to the first axis.

4. The system of claim 2, wherein the set of filter modules includes a set of excitation filters and a set of emission filters, including filters for at least two of:

HEX-based assays, FAM-based assays, ROX-based assays, and Cy5-based assays.

5. The system of claim 1, wherein the platform control module is further configured to angularly displace the platform about a third axis parallel to the broad surface of the platform to generate a distribution of focal lengths across the platform in relation to the optical sensor, and wherein the system further includes a processor configured receive a dataset from the optical sensor to calibrate at least one of the optical sensor and the optics manipulation module, based upon a focal length of the distribution of focal lengths providing a maximum contrast level.

6. The system of claim 1, wherein lens selector is a revolving nosepiece configured to receive a set of lenses, and wherein the translation stage is coupled to a plate surrounding the optical shaft coupled to the revolving nosepiece, thereby enabling vertical translation of the lens.

7. The system of claim 1, wherein the platform further includes a fluidic manifold coupled to a fluid source and configured to distribute a fluid to at least one imaging substrate, and wherein the system further includes a processor configured to facilitate analysis of real-time fluid flow at the at least one imaging substrate based upon data generated by the optical sensor.

8. The system of claim 1, further including at least one imaging substrate, in the set of imaging substrates, including an array of parallel pores configured to capture a set of cells in single-cell format.

9. The system of claim 8, further including a processor configured to translate a series of characters, physically located proximal to a pore of the array of parallel pores and detectable using the optical sensor, into a binary number indicative of an address of the pore.

10. A system for detecting biological sample features comprising:

a platform including a guide that holds an imaging substrate with a sample;

a filter module including an excitation filter and an emission filter;

a first light source configured to transmit light toward the sample at the platform from a first direction;

a second light source configured to transmit light through the excitation filter to reach the sample from a second direction opposed to the first direction;

a lens that focuses light from the excitation filter onto a target object of the sample, and transmits light from the target object, through the emission filter, toward an optical sensor superior to the filter module;

a platform control module configured to rotate the platform about a first axis perpendicular to a broad surface of the platform into a first configuration that aligns at least one imaging substrate between the first light source and the lens,

wherein the platform control module is further configured to angularly displace the platform about a third axis parallel to the broad surface of the platform to generate a distribution of focal lengths across the platform in relation to the optical sensor;

a filter stage, situated superior to the platform, configured to rotate the filter module about a second axis, parallel to the first axis, into a second configuration that positions the filter module to receive light from the second light source and to transmit light toward the optical sensor;

an optics manipulation module configured to vertically translate the lens; and

a processor including a first module that coordinates actuation by the platform control module, the filter stage, and the optics manipulation module and a second module that calibrates at least one of the optical sensor and the optics manipulation module to facilitate focusing of the target object, based upon the distribution of focal lengths.

11. The system of claim 10, further including a set of guides distributed about the platform in a first circular array and a set of filter modules distributed about the filter stage in a second circular array, wherein the platform control module is configured to rotate the platform into a first set of configurations that align each guide of the set of guides between the first light source and the lens, and wherein the filter stage is configured to rotate the set of filter modules into a second set of configurations that align each filter module of the set of filter modules to receive light from the second light source and transmit light toward the optical sensor.

12. The system of claim 11, wherein the optical sensor is superior to the filter module, which is superior to the platform, which is superior to the first light source.

13. The system of claim 12, wherein the platform control module includes an actuator configured to translate the platform along two directions perpendicular to the first axis.

14. The system of claim 11, wherein the set of filter modules includes a set of excitation filters and a set of emission filters, including filters for at least two of:

HEX-based assays, FAM-based assays, ROX-based assays, and Cy5-based assays.

15. The system of claim 10, wherein the optics manipulation module includes a lens selector coupled to the lens, an optical shaft coupled to the lens selector and concentrically aligned with the lens, and a translation stage coupled to the optical shaft, thereby enabling vertical translation of the lens.

16. The system of claim 10, wherein the filter module further includes dichroic mirror configured to bisect intersecting planes defined by the excitation filter and the emission filter, and wherein the system further includes a mirror configured to reflect light from the second light source into the filter module aligned in the second configuration, such that light from the second light source is reflected by 90°

twice before reaching the imaging substrate.

17. The system of claim 10, further including a tag identifying system configured to communicate information regarding the sample at the imaging substrate to the processor, and a thermal control module that controls thermal parameters of at least one of the imaging substrate and the sample.

18. The system of claim 10, wherein the platform further includes a fluidic manifold coupled to a fluid source and configured to distribute a fluid to the imaging substrate, and wherein the processor further includes a third module configured to facilitate analysis of real-time fluid flow at the imaging substrate based upon data generated by the optical sensor.

19. The system of claim 10, further including the imaging substrate, wherein the imaging substrate includes an array of parallel pores configured to capture a set of cells in single-cell format.

20. The system of claim 19, wherein the processor further includes a fourth module configured to translate a series of characters, physically defined proximal to a pore of the array of parallel pores and detectable using the optical sensor, into a binary number indicative of an address of the pore.