# US 10,499,797 B2 · System and method useful for sarcomere imaging via objective-based microscopy > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,499,797 B2 - Document ID: US10499797B2 - Status: Granted - Kind: B2 - Application number: 14/546,085 - Filed: 2014-11-18 - Published: 2015-05-21 - Granted: 2019-12-10 - Earliest claimed: 2007-07-03 - Assignees: The Board of Trustees of the Leland Stanford Junior University - Applicants: The Board of Trustees of the Leland Stanford Junior University - Inventors: Gabriel Nestor Sanchez; Scott L. Delp; Mark J. Schnitzer; Michael E. Llewellyn - Web page: https://patentgrep.com/patent/US10499797B2 ## CPC - A61B 5/4519: {Muscles (measuring muscular strength A61B5/224; using electromyography [EMG] A61B5/389)} - A61B 1/00045: {Display arrangement} - A61B 1/00165: {with light-conductive means, e.g. fibre optics (A61B1/07 takes precedence)} - A61B 1/0638: {providing two or more wavelengths} - A61B 1/313: for introducing through surgical openings, e.g. laparoscopes - A61B 5/0044: {for the heart} - A61B 5/0062: {Arrangements for scanning} - A61B 5/0084: {for introduction into the body, e.g. by catheters (A61B5/1459 takes precedence)} - A61B 5/6848: {Needles} - A61B 5/7207: {of noise induced by motion artifacts} - A61B 1/00172: {with means for scanning} - A61B 1/043: {for fluorescence imaging} - A61B 5/0068: {Confocal scanning} - A61B 5/0071: {by measuring fluorescence emission} - A61B 5/0086: {using infrared radiation} - A61B 5/4528: {Joints (A61B5/4533, A61B5/4538 take precedence)} - A61B 5/726: {using Wavelet transforms} - G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)} - G01N 21/6486: {Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence)} - A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D) - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract Biological tissue such as skeletal and cardiac muscle can be imaged by using an objective-based probe in the tissue and scanning at a sufficiently fast rate to mitigate motion artifacts due to physiological motion. According to one example embodiment, such a probe is part of a system that is capable of reverse-direction high-resolution imaging without needing to stain or otherwise introduce a foreign element used to generate or otherwise increase the sensed light. The probe can include a light generator for generating light pulses that are directed towards structures located within the thick tissue. The system can additionally include aspects that lessen adverse image-quality degradation. Further, the system can additionally be constructed as a hand-held device. ## Claims ### Claim 1 An apparatus for imaging a biological tissue of a subject, the apparatus comprising: a light source that is configured to provide light pulses; a scanning mirror in optical communication with the light source; an afocal lens arrangement including a fixed lens, wherein the afocal lens arrangement is configured to shift a focal plane at the biological tissue and is disposed between the light source and the scanning mirror, wherein the light source, scanning mirror and afocal lens arrangement are configured and arranged to direct the light pulses toward the biological tissue at a sufficiently fast line-resolution rate to mitigate motion artifacts due to contractile motion or physiological motion of the subject, which light pulses cause signals to be generated from and across at least a portion of the biological tissue; an objective in optical communication with the biological tissue and the light source, wherein the objective is configured and arranged to (i) focus the light pulses from the light source to the biological tissue, and (ii) collect the signals in a manner that lessens adverse image-quality degradation during changes of the focal plane at the biological tissue; a collection member, including circuitry, in optical communication with the biological tissue and the objective, wherein the collection member is configured and arranged to collect select ones of the generated signals that are present due to properties intrinsic to the biological tissue; and a computer processor that is coupled to the collection member, wherein the computer processor is configured to generate an artifact-mitigated image of the biological tissue by at least (i) directing the light source to provide the light pulses, (ii) directing the scanning mirror to send the light pulses toward the biological tissue at a line scan rate of at least 1 kHz, and (iii) using the select ones of the generated signals collected by the collection member to generate the artifact-mitigated image of the biological tissue. ### Claim 2 (depends on 1) The apparatus of claim 1, wherein at least a portion of the generated signals are second harmonic generation (SHG) signals, and wherein the computer processor is configured to image the biological tissue using the SHG signals. ### Claim 3 (depends on 1) The apparatus of claim 1, further comprising a probe in optical communication with the light source, wherein the probe is flexible. ### Claim 4 (depends on 1) The apparatus of claim 1, further comprising a probe in optical communication with the light source, wherein the probe is integrated with an electro-mechanical end portion that is configured and arranged to contact the biological tissue and, while contacting the biological tissue, electro-optically access the biological tissue. ### Claim 5 (depends on 4) The apparatus of claim 4, wherein the probe is a blunt probe or a needle. ### Claim 6 (depends on 1) The apparatus of claim 1, wherein the objective is a telecentric lens arranged in a light path in which the light pulses travel and is configured to maintain constant magnification during the changes of the focal plane. ### Claim 7 (depends on 1) The apparatus of claim 1, wherein the afocal lens arrangement is configured to maintain constant power of the light pulses generated from the light source. ### Claim 8 (depends on 1) The apparatus of claim 1, wherein the afocal lens arrangement is configured to maintain maximum resolution of the light pulses. ### Claim 9 (depends on 1) The apparatus of claim 1, wherein the afocal lens arrangement comprises a mobile lens, wherein the mobile lens is configured to shift the focal plane at the biological tissue. ### Claim 10 (depends on 1) The apparatus of claim 1, wherein the afocal lens arrangement is configured to provide convergence or divergence to the light pulses generated from the light source. ### Claim 11 (depends on 1) The apparatus of claim 1, wherein the objective is a lens configured to maintain constant magnification of the light pulses. ### Claim 12 (depends on 1) The apparatus of claim 1, wherein at least a portion of the generated signals are autofluorescence signals, and wherein the computer processor is configured to image the biological tissue using the autofluorescence signals. ### Claim 13 (depends on 4) The apparatus of claim 4, wherein the probe is configured to translate axially relative to the biological tissue. ### Claim 14 (depends on 5) The apparatus of claim 5, wherein the probe is a needle that is configured to puncture the biological tissue. ### Claim 15 (depends on 5) The apparatus of claim 5, herein the probe is a blunt probe that does not puncture the biological tissue. ### Claim 16 (depends on 5) The apparatus of claim 5, wherein the probe is a needle that is configured to translate axially relative to the biological tissue and to permit multiple independent measurements from a single injection. ### Claim 17 (depends on 5) The apparatus of claim 5, wherein the probe is a needle that has differing optical properties for wide field or high resolution imaging. ### Claim 18 (depends on 6) The apparatus of claim 6, wherein the objective is a lens configured to maintain constant magnification of the light pulses. ### Claim 19 (depends on 1) The apparatus of claim 1, further comprising an engageable clamp-mechanism interface that attaches a removable probe to the apparatus while maintaining optical alignment for image processing of the biological tissue. ### Claim 20 (depends on 1) The apparatus of claim 1, wherein the image of the biological tissue corresponds to at least 256 pixels spanning immediately adjacent z-lines of noncontracted sarcomere in the subject. ### Claim 21 (depends on 1) The apparatus of claim 1, further comprising an integrated power sensor in optical communication with the light source, the scanning mirror, and the afocal lens arrangement. ### Claim 22 (depends on 21) The apparatus of claim 21, wherein the integrated power sensor comprises a photodiode and a resistor. ### Claim 23 (depends on 22) The apparatus of claim 22, wherein the photodiode is configured and arranged to detect leakage light through a dichroic mirror. ### Claim 24 (depends on 1) The apparatus of claim 1, wherein the computer processor is further configured and arranged to regulate a power level of the light pulses at the biological tissue to prevent tissue damage. ### Claim 25 (depends on 1) The apparatus of claim 1, wherein the scanning mirror is disposed in a focal plane of the fixed lens.