	US 12,257,448 B2
	Image guided variable spot stimulation-based electrophysiology assessment device to determine changes in the functional health of biological samples during disease progression and therapy
Samarendra Kumar Mohanty, Southlake, TX (US); Sanghoon Kim, Bedford, TX (US); Michael Carlson, Rowlett, TX (US); and Subrata Batabyal, Arlington, TX (US)
Assigned to NANOSCOPE INSTRUMENTS, INC., Bedford, TX (US)
Filed by Nanoscope Instruments, Inc., Bedford, TX (US)
Filed on Nov. 16, 2023, as Appl. No. 18/511,752.
Claims priority of provisional application 63/383,979, filed on Nov. 16, 2022.
Prior Publication US 2024/0157169 A1, May 16, 2024
Int. Cl. A61N 5/06 (2006.01); A61B 3/10 (2006.01); A61B 5/00 (2006.01); A61B 5/398 (2021.01)

CPC A61N 5/0613 (2013.01) [A61B 3/102 (2013.01); A61B 5/398 (2021.01); A61B 5/4842 (2013.01); A61N 2005/0626 (2013.01); A61N 2005/0659 (2013.01)]

11 Claims

1. A device comprising:

i) an image-guided light stimulation beam allowing a variable spot size at a sample, for electrophysiological measurement of the sample,

ii) an image-guided light stimulation beam assembly comprising:

an imaging sub-assembly that provides illumination and collection of back-reflected light from the sample for imaging, and comprises near infrared (NIR) light from a low coherence source, and wherein the near infrared (NIR) light is able to be split into a sample beam and a reference beam for interferometric detection to obtain depth resolved images, and

a light stimulation sub-assembly comprising of light beams of different wavelengths with controllable intensities and/or pulsation rates,

iii) wherein a stimulation beam power at a sample plane ranges from 0.01 to 50 cd. s/m²for each individual stimulation light beam wavelength,

iv) wherein the light stimulation beam is combinable with the sample beam and able to be directed to the sample,

v) wherein the sample is selected from neurons or light-sensitive cells in-vitro or in-vivo,

vi) wherein the region of interest for the electrophysiological measurement on the sample in response to variable spot stimulation is pre-selected and controlled by scanning mirrors for deflecting the light stimulation beam(s),

vii) wherein the variable spot size is generated by a dynamic focusing element,

viii) wherein the light stimulation is able to be targeted to the pre-selected region of interest on the sample,

ix) wherein the light stimulation beam is able to be switched off during preselection of the region of interest for light stimulation, and wherein the preselection of the region of interest is based on morphologic/tomographic imaging,

x) wherein the sample beam for morphologic/tomographic imaging is deliverable via fiber-optic or free-space, and is able to be collimated, and deflected to the sample by the scanning mirrors and optical components,

xi) wherein the optical components are coated with an anti-reflection material for avoiding scattering and multiple reflections,

xii) wherein the back-reflected sample beam for morphologic/tomographic imaging from the sample is able to be routed back to a detector,

xiii) wherein a tomographic image is reconstructable by recording and analysis of interference between the back-reflected sample beam and the reference beam,

xiv) wherein the morphologic/tomographic image is able to be marked with selected regions of variable sizes to match with the light stimulation spots for electrophysiological measurements,

xv) wherein electrophysiological measurement is able to be conducted by electrodes connected to biosensing hardware,

xvi) wherein the image guided light stimulation beam, allowing the variable spot size at the sample, for electrophysiological measurement can be a benchtop system or wearable system,

xvii) wherein the wearable system for electrophysiological measurements comprises goggles or eye glasses, and

xviii) wherein the device is for generating spatial-temporal multiplexed variable-spot light stimulated electrophysiological measurements to enable improved signal to noise ratio from averaging and reducing measurement time, wherein:

i) the device is configured to stimulate a fraction of multiple non-overlapping spots among fractions of total number of spots of interest using a short burst of stimulation,

ii) the device is configured to reduce a time gap between stimulations by stimulating another fraction of multiple spots,

iii) the device is configured to change an order of synchronized stimulation among the fractions of the total number of spots of interest in each burst stimulation window,

iv) the device is configured to measure electrophysiological signals arising from a single set of bursts of stimulation containing linear combination of signal responses from the total number of spots of interest, and

v) the device is configured to solve linear equations based on location and time of stimulations and retrieve averaged signal from individual spots.