System and method useful for sarcomere imaging via objective-based microscopy
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.
1. A method for imaging an aspect of biological tissue including muscle via a light-delivering optical probe in the biological tissue, the method comprising:
using an objective during focal plane changes to focus the objective in the probe on the biological tissue while lessening adverse image-quality degradation;
using the optical probe to
send light pulses toward structure in the biological tissue at a sufficiently fast line-resolution rate to mitigate motion artifacts due to physiological motion,
cause, in response to the light pulses, signals to be generated from and across a sufficient portion of the structure to span a sarcomere length,
collect select ones of the generated signals that are predominantly present due to properties intrinsic to the structure; and
providing data in response to the collected signals for high-resolution imaging of said portion of the tissue structure.
2. The method of claim 1 , wherein the objective is telecentric and maintains constant magnification during the focal plane changes.
3. The method of claim 2 , wherein the telecentric objective maintains constant magnification of the optical probe.
4. The method of claim 1 , wherein the method further includes maintaining maximum resolution of the optical probe during the focal plane changes by providing the light pulses to overfill a back aperture of the optical probe.
5. The method of claim 1 , wherein the method further includes maintaining constant power of the optical probe during the focal plane changes by providing the light pulses with a constant beam waist.
6. The method of claim 1 , further comprising using an afocal lens arrangement to maintain power of the light pulses.
7. The method of claim 1 , further comprising using an afocal lens arrangement to provide convergence or divergence to the light pulses, thereby shifting the focal plane at the biological tissue.
8. The method of claim 7 , wherein the afocal lens arrangement includes a mobile lens and a fixed lens.
9. The method of claim 1 , wherein the light pulses provided to the optical probe are collimated and pivoted at a back aperture of the optical probe.
10. The method of claim 1 , wherein the optical probe includes an afocal lens arrangement that maintains maximum resolution of the optical probe.
11. The method of claim 1 , wherein the optical probe includes an afocal lens arrangement that maintains constant power of the optical probe.
12. The method of claim 1 , further including using an afocal lens arrangement to provide convergence or divergence to the light pulses.
13. A method for imaging an aspect of biological tissue including muscle via a light-delivering optical probe in the biological tissue, the method comprising:
providing the optical probe with a needle, the optical probe and needle being integrated with an electro-mechanical end portion that is configured and arranged to puncture the tissue and while in the tissue, electro-optically access the biological tissue;
using the optical probe to
send light pulses toward structure in the biological tissue,
cause, in response to the light pulses, signals to be generated from and across a sufficient portion of the structure in the biological tissue to span a sarcomere length,
in response to signals communicated between the electro-mechanical end portion and the biological tissue, collect selected ones of the generated signals that are predominantly present due to properties in the structure; and
providing data in response to the collected signals for high-resolution imaging of said portion of the tissue structure.
14. The method of claim 13 , wherein the light pulses are sent toward structure in the biological tissue at a sufficiently fast line-resolution rate to mitigate motion artifacts due to physiological motion.
15. The method of claim 13 , wherein the signals communicated between the electro-mechanical end portion and the biological tissue are sensed for muscular contraction.
16. The method of claim 13 , wherein the signals communicated between the electro-mechanical end portion and the biological tissue are the resulting sarcomere changes in response to an additional step of electrically stimulating the biological tissue.
17. The method of claim 13 , wherein the signals communicated between the electro-mechanical end portion and the biological tissue are to electrically stimulate.
18. The method of claim 13 , wherein the signals communicated between the electro-mechanical end portion and the biological tissue include signals which are electrically stimulating signals as well as responsive optically sensible signals.
19. The method of claim 18 , wherein the responsive optically sensible signals are useful for detecting muscular contraction and/or for measuring the resulting sarcomere changes.
20. The method of claim 13 , wherein the needle is translated axially relative to the tissue, enabling multiple independent measurements from a single injection.
21. The method of claim 13 , wherein the needle has differing optical properties for wide field or high resolution imaging.
22. The method of claim 13 , further including:
using an engageable clamp-mechanism interface to attach the optical probe with an imaging-processing microscope while maintaining optical alignment for imaging processing of the biological tissue.
23. The method of claim 13 , comprising:
providing a portable device for processing optical-signal data from the optical probe;
puncturing the tissue with the optical probe and needle; and
while in the tissue, collecting signals through the optical probe for high-resolution imaging of the tissue while lessening adverse image-quality degradation by controlling and maintaining power level and light-beam resolution for light pulsed through the light-delivering optical probe.
24. The method of claim 13 , wherein the portable device is hand-held.
25. The method of claim 13 , comprising:
puncturing the tissue with the optical probe and needle; and
while the optical probe is in the tissue, collecting signals for high-resolution imaging of the tissue while lessening adverse image-quality degradation by moving fluid near a biological-tissue image site for improving optical clarity.
26. The method of claim 25 , wherein moving fluid includes removing blood from the biological-tissue image site.
27. The method of claim 25 , wherein moving fluid includes providing saline to the biological-tissue image site.
28. The method of claim 25 , further including using the optical probe to
send light pulses toward structure in the biological tissue at a sufficiently fast line-resolution rate to mitigate motion artifacts due to physiological motion,
cause, in response to the light pulses, signals to be generated from and across a sufficient portion of the structure to span the sarcomere length,
collect selected ones of the generated signals that are predominantly present due to properties intrinsic to the structure; and
providing data in response to the collected signals for high-resolution imaging of said portion of the tissue structure.