Methods, systems, and devices for super resolution solid immersion lens microscopy
AssigneeSuRe Optics, Inc.
InventorsZong-Long Liau, Frederick J. Leonberger
Life of this patent
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Abstract
Described herein are methods and systems for the optical imaging of a physical specimen of interest that is in contact with, or in close proximity to, the backplane of a high refractive index solid-immersion lens (SIL), wherein the specimen comprises features of interest that act as a local high-refractive index regions. The SIL lens preferably comprises fiducial markers.
Claims48
A method of imaging a specimen containing features of interest with resolution in the sub-100 nm down to sub-nm range, using an annular solid immersion lens (ASIL), the method comprising the steps of:
- (a) directing electromagnetic radiation (emr) to the ASIL, using a laser scanning microscope (LSM), wherein:
- (i) the ASIL comprises a first surface comprising a convex region and a second surface that is substantially planar;
- (ii) at least a portion of the emr is transmitted through the convex region of the first surface of the ASIL, and propagates inside the ASIL toward the second surface;
- (iii) the portion of the emr transmitted through the convex region forms a focus near and/or on the second surface of the ASIL;
- and
- (iv) the second surface of the ASIL is in contact with a sample medium having a refractive index that is lower than the refractive index of the ASIL;
- (b) positioning a feature of interest of the specimen within the sample medium in close proximity to and/or in contact with the second surface of the ASIL, wherein the feature of interest has a refractive index that is higher than that of material immediately surrounding the feature of interest, and wherein the feature of interest is positioned within the focus formed by the emr, thereby illuminating the feature;
- (c) detecting emr that is either (i) transmitted from inside the ASIL into the sample medium, or (ii) reflected from inside the ASIL back toward the first surface of the ASIL;
- and
- (d) constructing an image of the specimen that resolves specimen features that are less than 100 nm using a LSM.
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A method of imaging a specimen containing features of interest with resolution in the sub-100 nm down to sub-nm range, using an annular solid immersion lens (ASIL), the method comprising the steps of:
- (a) directing electromagnetic radiation (emr) to the ASIL, wherein:
- (i) the ASIL comprises a first surface comprising a convex region, and a second surface that is substantially planar;
- (ii) at least a portion of the emr is transmitted through the convex region of the first surface of the ASIL, and propagates inside the ASIL toward the second surface;
- (iii) the portion of the emr transmitted through the convex region forms a focus near and/or on the second surface of the ASIL;
- and
- (iii) the second surface of the ASIL is in contact with a sample medium having a refractive index that is lower than the refractive index of the ASIL;
- (b) positioning specimen feature of interest of the specimen within the sample medium in close proximity to the second surface of the ASIL, wherein the feature of interest has a refractive index that is higher than that of material immediately surrounding the feature of interest, and wherein the feature of interest is positioned within the focus formed by the emr, thereby illuminating the specimen;
- (c) detecting fluorescent light that is emitted by at least one of (i) the feature of interest itself, and (ii) one or more secondary fluorescent objects within the sample medium;
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- (d) constructing an image that resolves specimen features that are less than 100 nm.
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A system for imaging a specimen containing features of interest with resolution in the sub-100 nm down to sub-nm range using an annular solid immersion lens (ASIL), the system comprising:
- a source of electromagnetic radiation (emr);
- an ASIL in contact with a sample medium and positioned in close proximity to, and/or in contact with, the specimen, the sample medium having a refractive index that is lower than the refractive index of the ASIL, the ASIL comprising:
- a first surface comprising a convex region, and
- a second surface that is substantially planar, wherein the second surface of the ASIL is in contact with the sample medium,
- such that at least a portion of the emr from the source is transmitted through the convex region of the first surface of the ASIL, and propagates inside the ASIL toward the second surface, the portion of the emr transmitted through the convex region forms a focus near and/or on the second surface of the ASIL, wherein a feature of interest of the specimen is in close proximity to the second surface of the ASIL, wherein the feature of interest has a refractive index that is higher than that of the sample medium, and wherein the feature of interest is positioned within the focus formed by the emr, thereby illuminating the feature of interest;
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- a detector for detecting emr that is either (i) transmitted from inside the ASIL into the sample medium, or (ii) reflected from inside the ASIL back toward the first surface of the ASIL.
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The system of claim 97, wherein the source, ASIL, and detector are part of a laser scanning microscope (LSM) and the source is aligned to direct the emr to the ASIL via optics of the LSM.
The system of claim 97, wherein the detector is mounted following the specimen and sample medium.
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Dependencies are inferred from the claim text.