Signal based sample preparation

Patent metadata

Field	Value
Publication number	US-10078037-B2
Application number	US-201314085653-A
Country	US
Kind code	B2
Filing date	Nov 20, 2013
Priority date	Nov 28, 2012
Publication date	Sep 18, 2018
Grant date	Sep 18, 2018

Abstract

Described is a system and method for detecting whether a biological event has occurred in a cellular sample, and then activating a fluidics system to fix the cell at the point in time with the event occurred. In one example, a sample preparation system includes a camera linked to a confocal microscope that is interrogating a cellular sample. Once a detectable event, such as a binding event, has occurred, the sample preparation system releases a fixative to fix the cell at the point in time when the event was detected.

First claim

What is claimed is:

1. A device, comprising:
an electronic hardware processor;
an electronic hardware memory, operably coupled to the electronic hardware processor, and storing:
an image analysis module comprising instructions stored in the electronic hardware memory, the instructions configuring the electronic hardware processor to analyze images provided by a light microscope to:
detect an individual cell in a biological sample, and
determine if a detectable biological event associated with the detected individual cell has occurred in the biological sample; and
a fluidics activation module comprising instructions that configure the electronic hardware processor to initiate treatment of the biological sample for electron microscopy in response to a determination that the detectable event has occurred based on the analyzed images associated with the light microscope.

2. The device of claim 1 , further comprising the light microscope.

3. The device of claim 1 , wherein the image analysis module further comprises instructions that configure the electronic hardware processor to determine a detectable event has occurred based on fluorescence in the biological sample.

4. The device of claim 1 , wherein instructions in the image analysis module configure the electronic hardware processor to read at least one image every microsecond.

5. The device of claim 1 , wherein the image analysis module further comprises instructions that configure the electronic hardware processor to determine if a detectable event has occurred based on detection of a fluorescent molecule.

6. The device of claim 5 , wherein the biological sample is label-free and the image analysis module further comprises instructions that configure the electronic hardware processor to determine the detectable event has occurred based on a fingerprint of the molecule in the form of a spectrum.

7. The device of claim 1 , wherein the image analysis module further comprises instructions that configure the electronic hardware processor to determine a detectable event has occurred based on quantitative phase contrast in the biological sample.

8. The device of claim 1 , wherein the image analysis module further comprises instructions that configure the electronic hardware processor to determine if a detectable event has occurred based on detection of receptor internalization; receptor binding; or membrane ion changes.

9. The device of claim 1 , wherein the image analysis module is further configured to:
detect a positon of a cell membrane for the detected cell;
detect when an antibody crosses a boundary of the detected cell membrane; and detect the biological event when the antibody crosses the boundary.

10. The device of claim 1 , wherein the image analysis module is further configured to detect when a fluorescently labeled polynucleotide reaches a nucleus of the individual cell, and to detect the biological event when the nucleus is reached by the labeled polynucleotide.

11. The device of claim 1 , wherein the biological sample is label-free, and the instructions stored in the electronic hardware memory configure the electronic hardware processor to analyze images provided by the light microscope to detect the individual cell in the label-free biological sample.

12. The device of claim 1 , wherein the fluidics activation module comprises instructions that configure the electronic hardware processor to initiate treatment of the biological sample for electron microscopy by applying a fixative to the biological sample in response to a determination that the detectable event has occurred.

13. The device of claim 1 , wherein the fluidics activation module comprises instructions that configure the electronic hardware processor to initiate treatment of the biological sample for electron microscopy by cryofixing the biological sample in response to a determination that the detectable event has occurred.

14. The device of claim 1 , the instructions configure the electronic hardware processor to:
analyze a plurality of images provided by a light microscope to detect individual cells in biological samples situated in respective sample wells; and
determine if detectable biological events associated with the detected individual cells have occurred in the biological samples in each of the sample wells; and
the fluidics activation module comprising instructions that configure the electronic hardware processor to initiate treatment of the biological samples for electron microscopy in each of the sample wells for which a corresponding detectable event has been established.

15. An electronic device, comprising:
a microscope configured to provide images of a biological sample; and
an electronic hardware processor configured to perform a method of activating a fluidics subsystem, the method comprising:
detecting an individual cell in an image of the biological sample obtained with the microscope;
analyzing the image to determine if a detectable biological event associated with the individual cell has occurred; and
initiating activation of the fluidics subsystem to prepare the biological sample for electron microscopy in response to detecting the biological event associated with the individual cell has occurred.

16. The electronic device of claim 15 , wherein the electronic hardware processor is configured to determine the detectable event has occurred based on fluorescence.

17. The electronic device of claim 15 , wherein the electronic hardware processor is configured to determine the detectable event has occurred based on detection of a fluorescent molecule.

18. The electronic device of claim 17 , wherein the electronic hardware processor is configured to determine the detectable event has occurred based on a fingerprint of the molecule in the form of a spectrum.

19. The electronic device of claim 15 , wherein the electronic hardware processor is further configured to determine a detectable event has occurred based on at least quantitative phase contrast in the biological sample.

20. The electronic device of claim 15 , wherein the electronic hardware processor is further configured to determine if a detectable event has occurred based on detection of receptor internalization; receptor binding; or membrane ion changes.

21. The electronic device of claim 15 , wherein the electronic hardware processor is further configured to determine a detectable event has occurred based on at least interference reflection in the biological sample.

22. The electronic device of claim 15 , wherein the electronic hardware processor is further configured to determine a detectable event has occurred based on at least Coherent anti-Stokes Raman scattering (CARS) microscopy in the biological sample.

23. The electronic device of claim 15 , wherein the electronic hardware processor is further configured to determine a detectable event has occurred based on at least second harmonic generation (SHG) in the biological sample.

24. The electronic device of claim 15 , wherein the electronic hardware processor is further configured to determine a detectable event has occurred based on at least third harmonic generation in the biological sample.

25. The electronic device of claim 15 , wherein the method the processor is configured to perform further includes detecting when a fluorescently labeled polynucleotide reaches a nucleus of the individual cell, and detecting the biological event when the nucleus is r

Assignees

• Fei Co

Inventors

• Lison Frank
• Greiser Jens
• Langhorst Matthias

Classifications

• G01N21/6458

  Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16) · CPC title

• G01N35/00594

  Quality control, including calibration or testing of components of the analyser · CPC title

• G01N1/30 Primary

  Staining; Impregnating {; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis} · CPC title

• G01N35/0092

  Scheduling · CPC title

• G01N1/312 Primary

  for samples mounted on planar substrates · CPC title