Live being optical analysis system and approach
View Patent ↗Analysis of live beings is facilitated. According to an example embodiment of the present invention, a light-directing arrangement such as an endoscope is mounted to a live being. Optics in the light-directing arrangement are implemented to pass source light (e.g., laser excitation light) into the live being, and to pass light from the live being for detection thereof. The light from the live being may include, for example, photons emitted in response to the laser excitation light (i.e., fluoresced). The detected light is then used to detect a characteristic of the live being.
1. A method for in vivo analysis of a freely-moving live being, the method comprising;
a) fixing a housing to the live being, wherein the housing encloses therein a light-directing arrangement, wherein the light-directing arrangement comprises a light source to provide stimulation light, and wherein a combined weight of the housing and the light-directing arrangement comprising the light source is less than 4 grams;
b) passing stimulation light through the light-directing arrangement;
c) using the light-directing arrangement, providing the stimulation light to a target portion of the freely-moving live being while the freely-moving live being moves about, wherein said stimulation light induces response light from said target portion;
d) passing the response light from the target portion to a light detector;
e) at the light detector, receiving and detecting the response light, and generating a signal corresponding to the detected light; and
f) generating a cellular resolution image of said target portion using said signal.
2. The method of claim 1 , wherein providing the stimulation light to the target portion includes selectively scanning stimulation light across the target portion.
3. The method of claim 1 , wherein fixing the housing includes fixing the housing to the live being for a period of days, wherein providing stimulation light includes providing stimulation light at several instances over the period of days and wherein receiving and detecting the response light includes receiving and detecting the response of the live being to the stimulation light provided at the several instances.
4. The method of claim 3 , further comprising observing outward behavior of the live being over the period of days and comparing the detected response to the outward behavior.
5. The method of claim 3 , further comprising applying a treatment condition to the live being, wherein detecting the response of the live being includes detecting a response of the live being to the treatment condition.
6. The method of claim 1 , wherein the live being is one or more of the following: a genetically engineered animal, a transgenic animal, an animal having undergone delivery of viral vectors that encode proteins, or an animal having undergone delivery of RNAi.
7. The method of claim 1 , wherein the live being expresses a fluorescent protein and wherein providing the stimulation light to the target portion includes stimulating the fluorescent protein, and wherein receiving and detecting the response light includes detecting fluoresced photons from the fluorescent protein.
8. The method of claim 1 , further comprising injecting the live being with a molecule that facilitates a fluorescent response to the provided light, and detecting the fluorescent response.
9. The method of claim 1 , wherein providing the stimulation light includes providing pulsed laser light at a wavelength selected to facilitate nonlinear excitation of a fluorochrome in the live being.
10. The method of claim 9 , wherein receiving and detecting the response light includes detecting photons as a non-linear response of the target portion.
11. The method of claim 9 , wherein providing the pulsed laser light includes providing laser light having a wavelength that limits the fluorochrome excitation to a point of focus of the provided laser light.
12. The method of claim 1 , wherein said generating a cellular resolution image comprises fluorescence imaging.
13. The method of claim 12 , wherein said fluorescence imaging is selected from one or more of the following: confocal fluorescence imaging, multi-photon fluorescence imaging, fluorescence lifetime imaging, fluorescence resonance energy transfer, or fluorescence recovery after photobleaching.
14. The method of claim 12 , wherein said generating a cellular resolution image comprises types of polarization sensitive imaging.
15. The method of claim 1 , comprising generating a cellular resolution image while said live being is freely moving about as under regular living conditions.
16. The method of claim 1 , wherein the live being is a rat, bird, or mouse.
17. The method of claim 16 , wherein the live being is a mouse.
18. The method of claim 17 , wherein the housing is fixed onto a head of the mouse.
19. The method of claim 1 , wherein said target portion comprises brain tissue.
20. The method of claim 1 , wherein said response light is limited to a limited point of focus of said stimulation light to said target portion.
21. The method of claim 1 , comprising passing said stimulation light to said target portion while said freely moving live being is moving about said live being's home setting.
22. The method of claim 1 , wherein said light-directing arrangement is operably coupled via wireless links to an external device.
23. The method of claim 22 , wherein the external device is a computer which is configured to selectively control one or more components of the light directing arrangement.
24. The method of claim 1 , wherein said generating said image of said target portion occurs while said freely moving live being is generally free to move about as would under regular living conditions.
25. The method of claim 1 , further comprising holding the light-directing arrangement via a coupling in a position relative to the live being.
26. The method of claim 1 , wherein the light-directing arrangement comprising the light source and enclosed in the housing further comprises the light detector.
27. The method of claim 1 , wherein the light-directing arrangement comprising the light source and enclosed in the housing further comprises a GRIN lens.
28. The method of claim 27 , wherein the GRIN lens is about 30-1000 μM in diameter.
29. The method of claim 1 , wherein the light-directing arrangement comprising the light source and enclosed in the housing further comprises a MEMS mirror.
30. The method of claim 1 , wherein the light-directing arrangement comprising the light source and enclosed in the housing further comprises an objective lens.
31. The method of claim 1 , wherein the light-directing arrangement comprising the light source and enclosed in the housing further comprises a dichroic mirror.
32. The method of claim 1 , wherein the housing has a dimension of about 35 mm or less.
33. The method of claim 1 , wherein the housing comprises a base plate and a protective cover that is selectively removable from the base plate.