阅读说明：本技术 一种兼顾电生理信号记录的微型荧光成像系统 (Miniature fluorescence imaging system considering electrophysiological signal recording ) 是由 裴为华 吴晓婷 王阳 于 2021-06-30 设计创作，主要内容包括：本公开提供了一种兼顾电生理信号记录的微型荧光成像系统,包括：一种微型荧光成像系统和电极探针。电极探针的第一端贴附于微型成像系统的成像透镜端面,且电极探针自成像透镜端面延伸贴附设置于成像透镜侧面；电极探针的第二端用于连接电生理采集设备。本公开将单一的光学信号检测系统改进为光学、电学信号同时检测的系统而不增加原有系统的体积、重量及应用复杂度等,可用于自由活动动物的皮层及深脑神经元活动的高时空分辨率的记录。(The present disclosure provides a micro fluorescence imaging system for electrophysiological signal recording, comprising: a micro fluorescence imaging system and an electrode probe. The first end of the electrode probe is attached to the end face of the imaging lens of the miniature imaging system, and the electrode probe extends from the end face of the imaging lens and is attached to the side face of the imaging lens; the second end of the electrode probe is used for connecting with an electrophysiological acquisition device. The present disclosure improves a single optical signal detection system into a system for simultaneously detecting optical and electrical signals without increasing the volume, weight, application complexity, etc. of the original system, and can be used for recording the cortical and deep brain neuron activities of freely moving animals with high spatial and temporal resolution.)

1. A micro fluorescence imaging system compatible with electrophysiological signal recording, comprising:

the first end of the electrode probe is attached to the end face of the imaging lens of the miniature imaging system, and the electrode probe extends from the end face of the imaging lens and is attached to the side face of the imaging lens;

and the second end of the electrode probe is used for being connected with an electrophysiological acquisition device.

2. The micro fluorescence imaging system compatible with electrophysiological signal recording of claim 1, wherein the electrode probe comprises:

the head part is used as a first end of the electrode probe and is attached to the end face of the imaging lens;

the first end of the neck is connected with the head, and the neck is bent for 90 degrees;

the first end of the wing part is connected with the second end of the neck part, and the wing part is attached to the side surface of the imaging lens; and

the tail part is used as the second end of the electrode probe, the first end of the tail part is connected with the second end of the wing part, and the second end of the tail part is connected with the electrophysiological acquisition device.

3. The micro fluorescence imaging system compatible with electrophysiological signal recording of claim 2, wherein the head has a diameter equal to a diameter of the end face of the imaging lens.

4. The electrophysiological signal recording enabled micro fluorescence imaging system of claim 2, wherein the electrode probe sequentially comprises, from bottom to top: a lower insulating layer, a metal layer, and an upper insulating layer.

5. The micro fluorescence imaging system compatible with electrophysiological signal recording of claim 4, wherein the metal layer comprises:

at least one gold electrode recording point for electrical signal recording of neurons of an imaging region, the gold electrode recording point corresponding to the head position;

at least one pad corresponding to the tail position; and

and two ends of the metal wire are respectively connected with the gold electrode recording point and the bonding pad.

6. The micro fluorescence imaging system compatible with electrophysiological signal recording of claim 5, further comprising:

and the PCB is connected with the welding disc in a welding way, and the electrophysiological acquisition device is connected with the PCB.

7. The micro fluorescence imaging system compatible with electrophysiological signal recording of claim 5, wherein the upper insulating layer comprises:

the first window is arranged on the upper insulating layer and corresponds to the position of the gold electrode recording point; and

and the second window is arranged on the upper insulating layer and corresponds to the position of the bonding pad.

8. The micro fluorescence imaging system for electrophysiological signal recording of claim 5, wherein the number of the gold electrode recording points is four.

9. The micro fluorescence imaging system for electrophysiological signal recording of claim 1 to 8, wherein the electrode probe is made of biocompatible flexible transparent material parylene C.

Technical Field

The present disclosure relates to the field of microelectronics, and more particularly to a micro fluorescence imaging system for electrophysiological signal recording.

Background

The research on brain function and neural circuits has very important significance for the diagnosis, regulation and treatment of brain diseases. The calcium imaging technology is a technology for reflecting the change of calcium ion concentration in neurons by using a calcium ion indicator, combines the change of an optical signal with the electrical activity behavior of the neurons, and can realize simultaneous monitoring of a large number of neuron states. The technology can provide very good spatial resolution imaging record and provides an effective technical means for analyzing the functional connection of the neurons. At present, the calcium imaging technology can be realized by a fluorescence microscope, a fiber-optic microscope, a two-photon microscope and the like, the two-photon microscope can realize three-dimensional depth imaging, and the two-photon microscope is widely applied to the calcium imaging technology. However, the calcium imaging technology has the disadvantage of poor time resolution, because the fluorescent calcium indicator has long quenching time after the fluorescence is excited, and the change of the fluorescent calcium indicator is difficult to follow the change of the action potential of high frequency.

The electrophysiological recording technology is a means for directly recording the rapid potential change of the neuron. This technique is still currently the gold standard for electrical tracing of neurons. However, this technique also has some disadvantages, for example, the number of channels of the electrode probe for recording electrophysiological signals is limited due to the size limitation, and in addition, the spatial resolution of this technique is limited because a single electrophysiological recording can only roughly determine the electrical activity of a certain region, cannot locate a specific neuron, and cannot interpret the functional connection between neurons.

Therefore, the combination of optical imaging technology and electrophysiological recording technology has become an effective tool for simultaneously observing and recording the neuronal activity of the brain in the field of neuroscience. At present, many research groups propose and prepare some cortical recording electrode probes aiming at the combination technology, and the cortical recording electrode probes are used for carrying out two-photon imaging while realizing cortical electroencephalogram recording. The current combination of simultaneous photoelectric recording is actually two separate bodies, i.e. electrode probes are laid on the cerebral cortex of an animal and then moved to a two-photon microscope for imaging, and usually the animal is fixed on a laboratory bench, so the research paradigm is limited.

The micro fluorescence imaging system is provided, the neuron activity observation and recording of freely movable animals are realized by the ultra-light weight and the extremely small volume, but the electrophysiological signals are difficult to realize synchronous acquisition and recording with fluorescence imaging in a non-fixed scene.

Disclosure of Invention

Technical problem to be solved

The present disclosure provides a micro fluorescence imaging system for electrophysiological signal recording to solve the above-mentioned technical problems.

(II) technical scheme

According to one aspect of the present disclosure, there is provided a micro fluorescence imaging system compatible with electrophysiological signal recording, comprising:

the first end of the electrode probe is attached to the end face of the imaging lens of the miniature imaging system, and the electrode probe extends from the end face of the imaging lens and is attached to the side face of the imaging lens;

and the second end of the electrode probe is used for being connected with an electrophysiological acquisition device.

In some embodiments of the present disclosure, the electrode probe includes:

the head part is used as a first end of the electrode probe and is attached to the end face of the imaging lens;

the first end of the neck is connected with the head, and the neck is bent for 90 degrees;

the first end of the wing part is connected with the second end of the neck part, and the wing part is attached to the side surface of the imaging lens; and

the tail part is used as the second end of the electrode probe, the first end of the tail part is connected with the second end of the wing part, and the second end of the tail part is connected with the electrophysiological acquisition device.

In some embodiments of the present disclosure, the diameter of the head is equal to the imaging lens end face diameter.

In some embodiments of the present disclosure, the electrode probe comprises, in order from bottom to top: a lower insulating layer, a metal layer, and an upper insulating layer.

In some embodiments of the present disclosure, the metal layer comprises:

at least one gold electrode recording point for electrical signal recording of neurons of an imaging region, the gold electrode recording point corresponding to the head position;

at least one pad corresponding to the tail position; and

and two ends of the metal wire are respectively connected with the gold electrode recording point and the bonding pad.

In some embodiments of the present disclosure, further comprising: and the PCB is connected with the welding disc in a welding way, and the electrophysiological acquisition device is connected with the PCB.

In some embodiments of the present disclosure, the upper insulating layer includes:

the first window is arranged on the upper insulating layer and corresponds to the position of the gold electrode recording point; and

and the second window is arranged on the upper insulating layer and corresponds to the position of the bonding pad.

In some embodiments of the present disclosure, the number of the gold electrode recording points is four.

In some embodiments of the present disclosure, the material of the electrode probe is biocompatible flexible transparent material parylene C.

(III) advantageous effects

According to the technical scheme, the micro fluorescence imaging system compatible with electrophysiological signal recording disclosed by the invention has at least one or part of the following beneficial effects:

according to the imaging lens, the specially designed electrode probe is integrated on the end face of the imaging lens, so that the imaging lens has an optical imaging function and an imaging area nerve signal recording function after being implanted, the size, the weight and the implantation complexity are not increased, and the photoelectric simultaneous recording of the nerve signals of freely moving animals is realized.

Drawings

FIG. 1 is a schematic view of a micro fluorescence imaging system for electrophysiological signal recording in accordance with an embodiment of the present disclosure.

FIG. 2 is a schematic structural diagram of the electrode probe in FIG. 1.

[ description of main reference numerals in the drawings ] of the embodiments of the present disclosure

1-an imaging device;

2-an imaging lens;

3-an electrode probe;

4-a lower insulating layer;

5-a metal layer;

6-upper insulating layer;

7-a head;

8-gold electrode recording dots;

9-neck part;

10-a wing portion;

11-tail.

Detailed Description

Aiming at the problems, the invention provides a micro fluorescence imaging system considering electrophysiological signal recording for simultaneously detecting nerve photoelectric signals, and integrates a specially designed electrode probe on the end surface of an imaging lens, so that the imaging lens has the functions of optical imaging and imaging area nerve signal recording after being implanted, the volume, the weight and the implantation complexity are not increased, and the nerve signal photoelectric simultaneous recording of freely moving animals is realized.

In a first exemplary embodiment of the present disclosure, a micro fluorescence imaging system is provided that allows for electrophysiological signal recording. FIG. 1 is a schematic view of a micro fluorescence imaging system for electrophysiological signal recording in accordance with an embodiment of the present disclosure. As shown in fig. 1, the present disclosure is a micro fluorescence imaging system for electrophysiological signal recording, comprising: an imaging device 1 and an electrode probe 3. An end portion of the imaging device 1 is provided with an imaging lens 2; the first end of the electrode probe 3 is attached to the end face of the imaging lens 2, and the electrode probe 3 extends from the end face of the imaging lens 2 and is attached to the side face of the imaging lens 2; the electrophysiological acquisition device is connected to the second end of the electrode probe 3. The present disclosure improves a single optical signal detection system into a system for simultaneously detecting optical and electrical signals without increasing the volume, weight, application complexity, etc. of the original system, and can be used for recording the cortical and deep brain neuron activities of freely moving animals with high spatial and temporal resolution.

The present disclosure is based on the design of a miniature imaging system, and can be integrated into the miniature imaging system, and a single optical probe is improved into an optical-electric dual-function probe without increasing the volume, weight and application complexity of the original system.

The respective components will be described in detail below.

As shown in fig. 2, the electrode probe 3 includes: head 7, neck 9, wings 10 and tail 11. The head part 7 serves as a first end of the electrode probe 3, the tail part 11 serves as a second end of the electrode probe 3, and the neck part 9 and the wing part 10 are sequentially arranged between the first end of the electrode probe 3 and the second end of the electrode probe 3. The head 7 has a contour which coincides with the end face of the lens, i.e. the diameter of the head 7 is equal to the diameter of the end face of the imaging lens 2. The head 7 is attached to the end face of the imaging lens 2. The contour of the neck portion 9 is not particularly limited, and may be a strip-like structure as shown in the drawings, or a contour structure thereof that meets the design requirements. The neck 9 can be bent by 90 ° so that the wing 10 is attached to the side of the imaging lens 2. The profile of the wing portion 10 is not limited in particular, and may be an oval structure in the figure, and generally, the surface area of the wing portion 10 should be larger than that of the neck portion 9 to better cover the side surface of the imaging lens 2. The wing part 10 is wrapped and attached to the lower side of the imaging lens 2. The tail 11 can be connected to the electrophysiological acquisition device.

Electrode probe 3 is the lamellar structure, includes in order from bottom to top: a lower insulating layer 4, a metal layer 5 and an upper insulating layer 6. The metal layer 5 includes: the gold electrode recording point 8, the pad and the metal wire, the gold electrode recording point 8 and the pad are connected at two ends of the metal wire, one end of the metal layer 5, where the gold electrode recording point 8 is arranged, corresponds to the head portion 7, and one end of the metal layer 5, where the pad is arranged, corresponds to the tail portion 11. In a specific embodiment four gold electrode recording dots 8 are provided, which are circular in shape. The number of the bonding pads is four corresponding to the gold electrode recording points 8, and the shape of the bonding pads is rectangular. Namely, the tail part 11 can lead out a metal lead and a bonding pad and is welded on a PCB board by gold wire ball to connect with the back end electrophysiological acquisition equipment. A first window and a second window are opened with respect to the upper insulating layer 6. Specifically, the first window is opened on the upper insulating layer 6, and corresponds to the position of the gold electrode recording point 8. The second window is opened on the upper insulating layer 6 and corresponds to the position of the pad.

The electrode probe is integrally made of biocompatible flexible transparent material parylene C, has good light transmittance, and does not influence the observation and collection of the micro imaging system on fluorescence signals.

The imaging device 1 is applied to calcium imaging recording of freely moving animals. Some research groups have developed lightweight miniature imaging devices that can be worn on the brains of freely moving animals for real-time imaging. The Miniscope is one kind of miniature imaging device, and mainly comprises a CMOS camera, an optical path system and a separable lens. The miniature imaging device can realize the recording of the brain waves of the cortex, namely the deep brain implantation and the multi-brain-area implantation recording.

In a first exemplary embodiment of the present disclosure, there is provided a method for preparing an electrode probe in a micro fluorescence imaging system compatible with electrophysiological signal recording, comprising:

step 1: designing the size and the outline of the electrode probe according to an imaging lens of the miniature imaging device;

step 2: preparing an electrode probe by using an MEMS (micro-electromechanical systems) process, wherein the process comprises the steps of growing a parylene C film by using a Chemical Vapor Deposition (CVD) method, photoetching, thermally evaporating metal, stripping, Reactive Ion Etching (RIE), electrolysis and the like;

and step 3: testing the light transmittance of the electrode probe substrate material by using a fiber optic spectrometer;

and 4, step 4: welding the electrode on a PCB by gold wire ball bonding to test the electrical characteristics of the electrode points;

and 5: the back of the head of the electrode probe with the PCB packaged is attached to the imaging end face of the lens by using transparent optical glue, and the neck of the electrode probe is bent upwards by 90 degrees so that the wing part is attached around the side face of the lens.

Step 6: and adhering the back of the wing part of the electrode probe of the packaged PCB to the side surface of the lens by using transparent optical glue.

So far, the embodiments of the present disclosure have been described in detail with reference to the accompanying drawings. It is to be noted that, in the attached drawings or in the description, the implementation modes not shown or described are all the modes known by the ordinary skilled person in the field of technology, and are not described in detail. Further, the above definitions of the various elements and methods are not limited to the various specific structures, shapes or arrangements of parts mentioned in the examples, which may be easily modified or substituted by those of ordinary skill in the art.

From the above description, those skilled in the art should clearly recognize that the present disclosure allows for a miniaturized fluorescence imaging system for electrophysiological signal recording.

In summary, the present disclosure provides a micro fluorescence imaging system for recording electrophysiological signals, in which a single optical probe is changed into a photoelectric probe for simultaneous detection and recording by optics and electricity, and calcium ion imaging in the hippocampus of a mouse and neuron activity electrical signal recording in an imaging region can be realized through multiple in vivo experiments.

It should also be noted that directional terms, such as "upper", "lower", "front", "rear", "left", "right", and the like, used in the embodiments are only directions referring to the drawings, and are not intended to limit the scope of the present disclosure. Throughout the drawings, like elements are represented by like or similar reference numerals. Conventional structures or constructions will be omitted when they may obscure the understanding of the present disclosure.

And the shapes and sizes of the respective components in the drawings do not reflect actual sizes and proportions, but merely illustrate the contents of the embodiments of the present disclosure. Furthermore, in the claims, any reference signs placed between parentheses shall not be construed as limiting the claim.

Unless otherwise indicated, the numerical parameters set forth in the specification and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by the present disclosure. In particular, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term "about". Generally, the expression is meant to encompass variations of ± 10% in some embodiments, 5% in some embodiments, 1% in some embodiments, 0.5% in some embodiments by the specified amount.

Furthermore, the word "comprising" does not exclude the presence of elements or steps not listed in a claim. The word "a" or "an" preceding an element does not exclude the presence of a plurality of such elements.

The use of ordinal numbers such as "first," "second," "third," etc., in the specification and claims to modify a corresponding element does not by itself connote any ordinal number of the element or any ordering of one element from another or the order of manufacture, and the use of the ordinal numbers is only used to distinguish one element having a certain name from another element having a same name.

Similarly, it should be appreciated that in the foregoing description of exemplary embodiments of the disclosure, various features of the disclosure are sometimes grouped together in a single embodiment, figure, or description thereof for the purpose of streamlining the disclosure and aiding in the understanding of one or more of the various disclosed aspects. However, the disclosed method should not be interpreted as reflecting an intention that: that is, the claimed disclosure requires more features than are expressly recited in each claim. Rather, as the following claims reflect, disclosed aspects lie in less than all features of a single foregoing disclosed embodiment. Thus, the claims following the detailed description are hereby expressly incorporated into this detailed description, with each claim standing on its own as a separate embodiment of this disclosure.

The above-mentioned embodiments are intended to illustrate the objects, aspects and advantages of the present disclosure in further detail, and it should be understood that the above-mentioned embodiments are only illustrative of the present disclosure and are not intended to limit the present disclosure, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present disclosure should be included in the scope of the present disclosure.