Target-locking single-molecule nanoscopy
US-2022163440-A1 · May 26, 2022 · US
US11567002B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-11567002-B2 |
| Application number | US-202017010802-A |
| Country | US |
| Kind code | B2 |
| Filing date | Sep 2, 2020 |
| Priority date | Oct 1, 2014 |
| Publication date | Jan 31, 2023 |
| Grant date | Jan 31, 2023 |
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A system for the identification of micro-organisms includes an irradiation unit adapted to sequentially provide coherent electromagnetic radiation of one or more wavelengths along a common optical path. A holder is adapted to retain a substrate having a surface adapted for growth of a micro-organism colony. A beamsplitter is adapted to direct the coherent electromagnetic radiation from the common optical path towards the retained substrate. An imager is arranged opposite the beamsplitter from the retained substrate and is adapted to obtain images of backward-scattered light patterns from the micro-organism colony irradiated by the respective wavelengths of the directed coherent electromagnetic radiation. Some examples provide radiation of multiple wavelengths and include an imager arranged optically downstream of the retained substrate to obtain images of forward-scattered light patterns from the micro-organism colony irradiated by the wavelengths of radiation. Organism identification methods are also described.
Opening claim text (preview).
The invention claimed is: 1. A system for the identification of micro-organisms, the system comprising: an irradiation unit that sequentially provides coherent electromagnetic radiation of multiple wavelengths along a common optical path; a holder adapted to retain a substrate having a solid surface adapted for holding a micro-organism colony, the microorganism colony comprising a plurality of microorganisms; a beamsplitter that directs the coherent electromagnetic radiation from the common optical path towards the retained substrate; a first imager arranged directly opposite the beamsplitter from the retained substrate and adapted to obtain images of elastically backward-scattered light patterns from the micro-organism colony irradiated by the respective wavelengths of the directed coherent electromagnetic radiation, the elastically backward-scattered light patterns formed substantially from a first portion of the directed coherent electromagnetic radiation scattered back from the micro-organism colony; a second imager arranged directly opposite the retained substrate from the beamsplitter and adapted to obtain an image of forward-scattered light patterns from the micro-organism colony irradiated by the respective wavelengths of the directed coherent electromagnetic radiation, the forward-scattered light patterns formed substantially from a second portion of the directed coherent electromagnetic radiation; and an optical density measurement unit that determines an optical density of the microorganism colony irradiated by the coherent electromagnetic radiation. 2. The system according to claim 1 , further comprising a stage that translates the retained substrate or the beamsplitter with respect to each other so that the directed coherent electromagnetic radiation irradiates the micro-organism colony. 3. The system according to claim 2 , further comprising a controller configured to: operate the stage and the irradiation unit to irradiate a first colony of a plurality of micro-organism colonies on the retained substrate; operate the first imager to obtain a first image and a second image of elastically backward-scattered light patterns from the first colony, the first image corresponding to a first wavelength and the second image corresponding to a second, different wavelength; subsequently, operate the stage and the irradiation unit to irradiate a second colony of the plurality of micro-organism colonies on the retained substrate; and operate the first imager to obtain a third image and a fourth image of elastically backward-scattered light patterns from the second colony, the third image corresponding to a third wavelength and the fourth image corresponding to a fourth wavelength different from the third wavelength. 4. The system according to claim 3 , wherein the controller is configured to utilize at least one the first portion, the second portion, and the optical density to measure a cross-sectional morphology of the micro-organism colony. 5. The system according to claim 3 , wherein the controller utilizes the optical density measurement unit to determine an aspect ratio of the microorganism colony. 6. The system according to claim 3 , wherein the controller utilizes the optical density measurement unit to determine an elevation height of a center of the microorganism colony. 7. The system according to claim 1 , wherein the irradiation unit comprises: multiple sources for the respective wavelengths of the coherent electromagnetic radiation; and one or more source beamsplitters configured to direct the coherent electromagnetic radiation from the sources to the common optical path. 8. The system according to claim 7 , wherein the source beamsplitters comprise respective pellicle beamsplitters. 9. The system according to claim 1 , wherein the irradiation unit further comprises a sensor configured to detect a level value of the coherent electromagnetic radiation. 10. The system according to claim 9 , wherein the controller is further configured to: determine respective level values of the multiple wavelengths using the sensor; and adjust respective output levels of the coherent electromagnetic radiation of the respective ones of the wavelengths based at least in part on the respective level values and a selected set point. 11. The system according to claim 10 , further comprising: a first sensor arranged optically upstream of the retained substrate and configured to detect a first level value of the coherent electromagnetic radiation; and a second sensor arranged optically downstream of the retained substrate and configured to detect a second level value of the coherent electromagnetic radiation. 12. The system according to claim 1 , wherein the second portion formed from the forward-scattered light patterns and the optical density of the microorganism colony irradiated by the coherent electromagnetic radiation are measured simultaneously. 13. A system for the identification of micro-organisms, the system comprising: an irradiation unit that provides coherent electromagnetic radiation of a selected wavelength along an optical path; a holder adapted to retain a substrate having a surface adapted for growth of a micro-organism colony, the microorganism colony comprising a plurality of microorganisms; a beamsplitter that directs the coherent electromagnetic radiation from the optical path towards the retained substrate; a first imager arranged directly opposite the beamsplitter from the retained substrate and adapted to obtain an image of an elastically backward-scattered light pattern from the micro-organism colony irradiated by the directed coherent electromagnetic radiation, the backward scattered light patterns formed substantially from a first portion of the directed coherent electromagnetic radiation scattered back from the micro-organism colony; a second imager arranged directly opposite the retained substrate from the beamsplitter and adapted to obtain an image of forward-scattered light patterns from the micro-organism colony irradiated by the directed coherent electromagnetic radiation, the forward-scattered light patterns formed substantially from a second portion of the directed coherent electromagnetic radiation; and an optical density measurement unit that determines an optical density of the microorganism colony irradiated by the coherent electromagnetic radiation. 14. The system according to claim 13 , further comprising: a stage adapted to translate the retained substrate or the beamsplitter with respect to each other so that the directed coherent electromagnetic radiation irradiates the micro-organism colony; and a controller configured to: operate the stage and the irradiation unit to successively irradiate ones of a plurality of micro-organism colonies on the retained substrate; and operate the first imager to obtain a plurality of images of elastically backward-scattered light patterns from the successively-irradiated micro-organism colonies, the plurality of images including at least first and second images of a first colony at respective, different wavelengths, and third and fourth images of a second, different colony at respective, different wavelengths. 15. The system according to claim 13 , wherein: the irradiation unit further comprises a sensor configured to detect a level value of the coherent electromagnetic radiation; and the system further comprises a controller responsive to the level value and a selected set point to adjust an output level of the coherent electromagnetic radiation. 16. A system for the identification of micro-organis
inside a container, e.g. in an ampoule (G01N21/53 takes precedence) · CPC title
Scattering, i.e. diffuse reflection (G01N21/25, G01N21/41 take precedence {G01N21/55 takes precedence}) · CPC title
Details, e.g. use of specially adapted sources, lighting or optical systems · CPC title
Forward scatter; Low angle scatter · CPC title
Backscatter · CPC title
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