JP2003164416A - Method and apparatus for obtaining fluorescence spectrum - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide, at high S/N, data on an intensity distribution of the spectrum of fluorescence generated from living tissue, using a method and apparatus for obtaining fluorescence spectra. <P>SOLUTION: Light caused to impinge through a light propagation probe 5 when the living tissue 1 is irradiated with excitation light emitted from an LD light source 61 is subjected to spectrophotometry a plurality of times by a spectrophotometry means 10 so as to obtain a plurality of fluorescence spectrum intensity distribution data. A dispersion computing means 15 computes the dispersion of the data according to values involved in the data, and based on the dispersion, a contact determining means 20 determines whether or not the probe 5 is in contact with the living tissue 1; when the probe 5 is determined to be in contact with the tissue 1, the fluorescence spectrum intensity distribution data obtained at that time is employed to obtain data on the spectrum intensity distribution of the fluorescence at high S/N. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a fluorescence spectrum acquisition method and apparatus, and more particularly to a fluorescence spectrum acquisition method and apparatus for spectrophotometrically injecting fluorescence generated from living tissue by excitation light irradiation through a light propagation probe. It is a thing.

[0002]

2. Description of the Related Art Conventionally, a light propagation probe in which an optical fiber is connected to a glass sphere has been used, and after inserting the light propagation probe into a living tissue, the glass sphere portion is brought into contact with the living tissue and irradiated with excitation light. Fluorescence generated from the biological tissue is made incident from the glass sphere and propagated through the optical fiber, and the fluorescence propagated through the optical fiber is spectrophotometrically measured to measure the spectrum of the fluorescence generated from the biological tissue. The device is known.

In such an apparatus, in order to improve the measurement accuracy of the fluorescence spectrum to be acquired, the spectral intensity distribution data of a plurality of fluorescence obtained by a plurality of spectrophotometric measurements are added and averaged, and the fluorescence generated from the above-mentioned biological tissue. Has acquired the spectrum of.

[0004]

However, when the fluorescence spectrum is measured, the glass bulb is not in stable contact with the living tissue but is simply in contact therewith, so that the living tissue is not contacted. During the measurement of the fluorescence emitted from the tissue (hereinafter referred to as tissue fluorescence), the positional relationship between the living tissue and the glass bulb portion (the gap between them, the angle, etc.) may become unstable. When the positional relationship becomes unstable, the mucus that covers the living tissue invades between the living tissue and the glass sphere portion, and the spectral intensity distribution different from the tissue fluorescence generated from the mucus under the irradiation of excitation light is generated. Fluorescence that becomes noise (hereinafter referred to as mucus fluorescence) is incident on the light propagation probe together with the tissue fluorescence and spectrophotometrically measured to obtain S /
The tissue intensity spectrum intensity distribution data of low N may be acquired.

Therefore, during a plurality of measurements, the above-mentioned positional relationship becomes unstable and the glass bulb part does not contact the living tissue, or the area where the glass bulb part contacts the living tissue. When the area of the tissue becomes larger or smaller, the ratio of the amounts of tissue fluorescence and mucus fluorescence, which are simultaneously incident on the light propagation probe and spectrophotometrically changed, changes. There is a problem in that a plurality of acquired spectral intensity distribution data have variations, and it is difficult to obtain tissue fluorescence spectral intensity distribution data with high S / N even if these spectral intensity distribution data are added and averaged.

The present invention has been made in view of the above circumstances, and provides a fluorescence spectrum acquisition method and apparatus capable of acquiring spectrum intensity distribution data of fluorescence generated from living tissue with a high S / N. The purpose is.

[0007]

A first method for obtaining a fluorescence spectrum of the present invention is that fluorescence generated from living tissue by irradiation of excitation light is made incident through a light propagation probe, and the fluorescence made incident through this light propagation probe is dispersed. In the fluorescence spectrum acquisition method of photometrically acquiring the spectral intensity distribution data of the fluorescence, a plurality of optical spectra are obtained by spectrophotometrically measuring the light incident through the light propagation probe while the excitation light is being applied to the biological tissue. Obtaining the intensity distribution data, obtain the variation of the plurality of optical spectrum intensity distribution data from the values of the plurality of optical spectrum intensity distribution data, whether the light propagation probe is in contact with the biological tissue based on this variation It is characterized by making a judgment.

The second fluorescence spectrum acquisition method of the present invention is such that fluorescence generated from living tissue by irradiation of excitation light is made incident through a light propagation probe, and the fluorescence made incident through this light propagation probe is spectrophotometrically measured to obtain this fluorescence. In the fluorescence spectrum acquisition method for acquiring the spectrum intensity distribution data of, when the living tissue is illuminated by the background light, and the excitation light is not applied to the living tissue, a plurality of the background lights incident through the light propagation probe are A plurality of background light intensity data representing the intensity of this background light is acquired by performing photometry, the variations of these background light intensity data are obtained from the values of the above-mentioned background light intensity data, and the light is propagated based on this variation. It is characterized by determining whether or not the probe is in contact with the living tissue.

The first fluorescence spectrum acquisition apparatus of the present invention comprises a light-propagation probe for injecting fluorescence generated from living tissue by irradiation of excitation light, and spectrophotometric measurement of the fluorescence incident through the light-propagation probe to obtain the fluorescence. In the fluorescence spectrum acquisition device having a spectrophotometric means for acquiring the spectral intensity distribution data of the spectrophotometric means, the spectrophotometric means, when the excitation light is applied to the biological tissue, makes the light incident through the light propagation probe a plurality of times. This is to obtain a plurality of light spectrum intensity distribution data by spectrophotometry, and calculate the variation of the plurality of light spectrum intensity distribution data based on the values of the plurality of light spectrum intensity distribution data obtained by this spectrophotometric means. Variation calculation means for
It is characterized by further comprising contact determination means for determining whether or not the light propagation probe is in contact with the biological tissue based on the variation calculated by the variation calculation means.

Further, the variation calculating means calculates the variation based on the difference between the profiles of the spectrum intensity distributions represented by the plurality of optical spectrum intensity distribution data, or calculates the light incident through the light propagation probe a plurality of times. The variation may be calculated based on a difference between two continuous data among a plurality of optical spectrum intensity distribution data obtained by spectrophotometry.

In the fluorescence spectrum acquisition device, when the excitation light is intermittently irradiated in the presence of the background light, the spectrophotometric means emits the light incident through the light propagation probe while the excitation light is irradiated. The background light spectrum obtained by spectrophotometrically measuring the background light incident through the light propagation probe when the excitation light is not irradiated following the irradiation of the excitation light from the light spectrum intensity distribution data obtained by spectrophotometric measurement It is preferable that the background light spectrum intensity distribution data representing the intensity distribution is subtracted to obtain the light spectrum intensity distribution data used for calculating the variation.

Further, the fluorescence spectrum acquisition device may be provided with display means for displaying the values of a plurality of fluorescence spectrum intensity distribution data used for calculating the variation.

The second fluorescence spectrum acquisition apparatus of the present invention comprises a light-propagation probe for injecting fluorescence generated from living tissue by irradiation of excitation light, and spectrophotometric measurement of fluorescence incident through the light-propagation probe to obtain the fluorescence. In a fluorescence spectrum acquisition device equipped with a spectrophotometric means for acquiring the spectral intensity distribution data of the background, the background incident through the light propagation probe when the living tissue is illuminated by the background light and the excitation light is not applied to the living tissue. A background light intensity metering unit that measures a plurality of times of light to obtain a plurality of background light intensity data representing the intensity of the background light, and a variation of the plurality of background light intensity data based on the plurality of background light intensity data. Variation calculating means for calculating, and the light propagation probe based on the variation calculated by this variation calculating means And it is characterized in that it comprises a contact determination means for determining whether or not in contact.

The variation calculating means calculates the variation based on the difference of the plurality of background light intensity data, or measures the background light incident through the light propagation probe a plurality of times. Of the background light intensity data, the above variation may be calculated based on the difference between two consecutive data.

Further, the fluorescence spectrum acquisition apparatus may be provided with display means for displaying the values of the plurality of background light intensity data.

Further, the fluorescence spectrum acquisition apparatus may be provided with an informing means for informing the result of the determination by the contact determining means.

The variations of the plurality of background light intensity data are, for example, a value (standard deviation, etc.) representing a statistically determined distribution state of these data, a maximum value of a difference between these data, It is meant to represent the distribution of these data (including the distribution shown by the difference between two data), such as the difference between continuously acquired data.

Further, the variation of the plurality of optical spectrum intensity distribution data is a distribution (2 which is shown based on a difference in profile shape of these optical spectrum intensity distributions.
(Including distributions indicated by the difference in the shape of the two profiles).

Further, "the light propagation probe is in contact with the living tissue" means that substantially the entire area of the incident surface of the light propagation probe on which the spectrophotometric light is incident is in close contact with the living tissue,
This means a state in which there is almost no air or mucus covering the biological tissue in the region sandwiched between the biological tissue and the light propagation probe in the optical path through which the spectrophotometric light is propagated.
Hereinafter, the above state will be referred to as a “predetermined contact state”.

The background light means light that illuminates living tissue with a constant intensity, such as outdoor light, indoor illumination light, and illumination light that illuminates living tissue. May illuminate the biological tissue through the epidermis of the biological body.

The "background light incident through the light propagation probe when the excitation light is not irradiated subsequent to the irradiation of the excitation light" means that the excitation light continued before the irradiation of the excitation light. The background light may be the background light that is incident when the excitation light is not irradiated, or the background light that is incident when the excitation light is not irradiated subsequent to the irradiation of the excitation light.

[0022]

The present invention has been accomplished based on the following findings.

In spectrophotometry in which tissue fluorescence is made incident through a light propagation probe to obtain spectral intensity distribution data, the living tissue and the light propagation probe are contacted with each other only when the light propagation probe comes into contact with the living tissue in a predetermined contact state. The positional relationship of is stable. Therefore, a plurality of spectral intensity distribution data acquired by a plurality of measurements in this predetermined contact state, there is little variation in the ratio of the amount of light of the tissue fluorescence and the mucus fluorescence incident from the light propagation probe and spectrophotometrically dispersed. The S / N ratio is suppressed to a low level, and intrusion of mucus between the biological tissue and the light propagation probe is prevented, so that the mixture of mucus fluorescence, which is noise incident from the light propagation probe, is reduced. This is the spectrum intensity distribution data of the tissue fluorescence that it has.

On the other hand, when the light propagation probe is not in contact with the living tissue in a predetermined contact state and the positional relationship between the living tissue and the light propagation probe is not stable, the tissue fluorescence spectrophotometrically measured as described above Since the ratio of the amount of light to the mucus fluorescence changes for each measurement, the variation of the acquired plurality of spectral intensity distribution data becomes large.

That is, when there are few variations in the acquired plurality of spectral intensity distribution data, it can be considered that the light propagation probe is in contact with the living tissue in a predetermined contact state, and at this time, mucus fluorescence which becomes noise is mixed. Tissue fluorescence is spectrophotometrically measured in a state in which there is little.

Therefore, the plurality of spectrum intensity distribution data obtained when the variation of the plurality of acquired spectrum intensity distribution data is small can be adopted as the spectrum intensity distribution data of the tissue fluorescence having a high S / N.

When the living tissue is not irradiated with the excitation light, the background light illuminating the living tissue is made incident through the light propagation probe, and the intensity of the background light made incident through the light propagation probe is measured. Even in this case, the positional relationship between the biological tissue and the light propagation probe is stabilized only when the light propagation probe is brought into contact with the biological tissue in a predetermined contact state, that is, the mucus between the biological tissue and the light propagation probe. The amount of background light reflected by the boundary surface between biological tissue and mucus or the boundary surface between mucus and air is kept constant (the amount of incident light is extremely small), which is prevented by the penetration of air and air. The variation in the plurality of background light intensity data acquired by the plurality of measurements in the predetermined contact state can be suppressed.

On the other hand, when the light propagation probe is not in contact with the living tissue in a predetermined contact state and the positional relationship between the living tissue and the light propagating probe is not stable, the mucus is placed between the living tissue and the light propagating probe. And air enter, and the state of mucus and air entering is not stable, so the background light reflected by the interface between biological tissue and mucus or the interface between mucus and air is incident on the light propagation probe. The light amount is not stable, and the variations in the plurality of background light intensity data acquired by the plurality of measurements increase.

That is, when the variation in the acquired background light intensity data is small, it can be considered that the light propagation probe is in contact with the living tissue in a predetermined contact state, and the light propagation probe is acquired at a time close to the photometric time of the background light. The acquired spectral intensity distribution data can also be regarded as acquired when the light propagation probe is in contact with the biological tissue in a predetermined contact state.

Therefore, the spectral intensity distribution data acquired by spectrophotometry at a time close to the time when a plurality of background light intensity data with little variation are acquired is converted into a high S / N ratio.
It can be used as the spectral intensity distribution data of the tissue fluorescence obtained in.

According to the first fluorescence spectrum acquisition method and apparatus of the present invention, a plurality of optical spectra are obtained by spectrophotometrically measuring the light incident through the light propagation probe a plurality of times while the living tissue is irradiated with the excitation light. Obtaining the intensity distribution data, obtain the variation of the plurality of optical spectrum intensity distribution data from the values of the plurality of optical spectrum intensity distribution data, whether the light propagation probe is in contact with biological tissue based on this variation. I decided to make a decision,
A plurality of light spectrum intensity distribution data acquired when it is determined that the light propagation probe is in contact with the biological tissue in a predetermined contact state can be adopted as the light spectrum intensity distribution data of the tissue fluorescence, whereby The spectral intensity distribution data of tissue fluorescence can be acquired with a high S / N.

Further, the variation calculating means calculates the variation based on the difference of the profiles of the spectrum intensity distributions represented by the plurality of optical spectrum intensity distribution data, or splits the light incident through the light propagation probe a plurality of times. Among the plurality of light spectrum intensity distribution data obtained by photometry, if the above-mentioned variation is calculated based on the difference between two consecutive data, the light propagation probe contacts the biological tissue. It is possible to more reliably determine whether or not there is.

Further, the spectrophotometric means spectrophotometrically measures the light incident through the light propagation probe while the excitation light is being radiated, and the excitation light following the irradiation of the excitation light is obtained from the optical spectrum intensity distribution data obtained. The background light spectrum intensity distribution data representing the background light spectrum intensity distribution obtained by spectrophotometrically measuring the background light incident through the light propagation probe when not irradiated is subtracted from the background light spectrum intensity distribution data to be used in the calculation of the above-mentioned variation. If the distribution data is acquired, the above variation is calculated based on the optical spectrum intensity distribution data in which less noise is mixed, so it is possible to more reliably determine whether or not the light propagation probe is in contact with the biological tissue. can do.

If a display means for displaying a plurality of values of the fluorescence spectrum intensity distribution data used for calculating the variation is further provided, the variation of the light spectrum intensity distribution data can be visually confirmed.
The variation can be calculated more reliably.

According to the second fluorescence spectrum acquisition method and apparatus of the present invention, when the living tissue is illuminated by the background light and the living tissue is not irradiated with the excitation light,
A plurality of background light intensity data representing the intensity of the background light is obtained by measuring the background light incident through the light propagation probe a plurality of times, and the plurality of background light intensity data is obtained from the values of the plurality of background light intensity data. Since it is determined whether the light propagation probe is in contact with the biological tissue based on this variation, it is determined that the light propagation probe is in contact with the biological tissue in a predetermined contact state. It is possible to employ a plurality of spectral intensity distribution data obtained at a time close to the time point as the tissue fluorescence spectral intensity distribution data, and thereby obtain the tissue fluorescence spectral intensity distribution data with a high S / N. it can.

Further, the variation calculating means calculates the variation based on the difference between the plurality of background light intensity data,
For example, if the background light intensity data obtained by measuring the background light incident through the light propagation probe a plurality of times is calculated based on the difference between two consecutive data, It is possible to more reliably determine whether or not the light propagation probe is in contact with the living tissue.

Further, if a display means for displaying a plurality of values of the background light intensity data is further provided, the variation of the background light intensity data can be visually confirmed, and the above variation can be calculated more reliably. can do.

Further, by further providing an informing means for informing the result of the determination by the contact determining means, it is possible to more surely recognize whether or not the light propagation probe is in contact with the living tissue.

[0039]

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described below with reference to the drawings. FIG. 1 is a block diagram showing a schematic configuration of the fluorescence spectrum acquisition apparatus according to the first embodiment, FIG. 2 is a diagram showing a state where a light propagation probe is in contact with a biological tissue in a predetermined contact state, FIG. 3 and FIG. 5 is a diagram showing a first variation calculation method in the first embodiment, FIG. 4 is a diagram showing a state where the light propagation probe is not in contact with a biological tissue in a predetermined contact state, and FIGS. 6 and 7 are First
It is a figure which shows the calculation method of the 2nd variation in embodiment of this.

The fluorescence spectrum acquisition apparatus 101 according to the first embodiment of the present invention is generated from the living tissue 1 by the LD light source 61 which emits the excitation light of wavelength 410 nm and the irradiation of the excitation light which is emitted from the LD light source 61. The light propagating probe 5 for injecting the fluorescent light, the spectrophotometric means 10 for spectrophotometrically measuring the fluorescent light incident through the light propagating probe 5 to obtain the spectral intensity distribution data of the fluorescence, and the incident light through the light propagating probe 5. The fluorescence propagation optical system 62 for making the fluorescence enter the spectrophotometric means 10 and a part of the fluorescence propagation optical system 62, and the fluorescence made incident on the light propagation probe 5 is emitted from the side where the light propagation probe 5 is emitted. An excitation light propagation optical system 63 for making the excitation light incident on the light propagation probe 5 is provided.

The light propagation probe 5 comprises a glass sphere portion 6 and an optical fiber portion 7 connected to the glass sphere portion 6, and L
The excitation light emitted from the D light source 61 is the excitation light propagation optical system 6
3 is emitted from the glass sphere portion 6 toward the living tissue 1 through the optical fiber portion 7, and the tissue fluorescence generated from the living tissue 1 by the irradiation of the excitation light is made incident from the glass sphere portion 6 and the optical fiber portion 7 and the fluorescence. The light enters the spectrophotometer 10 through the propagation optical system 62.

The excitation light propagating optical system 63 collimates the excitation light emitted from the LD light source 61 into parallel light.
0, a half mirror 71 that transmits and reflects the parallel excitation light, a first convex lens 72 that collects the excitation light reflected by the half mirror 71, and an excitation light that is collected by the first convex lens 72 Pump power controller 73 for controlling the power of the LD light source 61 so that the intensity of the pump light is constant, and a wavelength of 440 nm or more for reflecting the pump light transmitted through the half mirror 71 and a wavelength for transmitting the pump light. It has a dichroic mirror 74 that reflects light of less than 440 nm, and a second convex lens 75 that collects the excitation light reflected by the dichroic mirror 74 and makes it incident on the end face T of the optical fiber section 7.

The fluorescence propagation optical system 62 includes the light propagation probe 5
The second convex lens 75, which collimates the light that is incident on the light propagation probe 5 through the glass sphere portion 6 and is emitted from the end surface T of the optical fiber portion 7 into parallel light, is converted into parallel light by the dichroic mirror. 74 by 44
Third convex lens 7 that collects the parallel light that has transmitted only the wavelength component of 0 nm or more and makes it enter the spectrophotometer 10.
Have six.

The spectrophotometer 10 spectrophotometrically measures the light incident through the light propagation probe 5 a plurality of times while the living tissue 1 is being irradiated with the excitation light, and obtains a plurality of optical spectrum intensity distribution data. It is a thing.

Further, this fluorescence spectrum acquisition apparatus 10
1 is a variation calculation means 15 for calculating the variation of the plurality of optical spectrum intensity distribution data based on the values of the plurality of optical spectrum intensity distribution data acquired by the spectrophotometric means 10, and the variation calculation means 15. The contact determination means 20 for determining whether or not the light propagation probe 5 is in contact with the living tissue 1 based on the determined variation, the notification means 25 for notifying the result of the determination by the contact determination means 20, and the light propagation probe 5. And an averaging unit 30 that calculates the tissue fluorescence spectrum intensity distribution data by averaging the plurality of optical spectrum intensity distribution data obtained when it is determined that the tissue is in contact with the biological tissue 1.
Display timing of operation in each of the above-mentioned elements and display means 35 for displaying the tissue fluorescence spectrum intensity distribution data and displaying the values of a plurality of optical spectrum intensity distribution data used for calculation of the variations output from the variation computing means 15. And a controller (not shown) for controlling the above.

The variation calculation means 15 calculates the variation based on the difference between the profiles of the spectrum intensity distributions represented based on the plurality of optical spectrum intensity distribution data, or calculates the plurality of optical spectrum intensity distribution data. Among them, the variation is calculated based on the difference between two continuous data.

Next, the operation of the above embodiment will be described.

After inserting the light propagation probe 5 into the living body and bringing the glass sphere portion 6 into contact with the living tissue 1, the LD light source 61 is turned on and the excitation light emitted from the LD light source 61 is excited by the excitation light propagation optical system. 63, optical fiber portion 7 and glass bulb portion 6
The biological tissue 1 is irradiated through the through. The tissue fluorescence generated from the living tissue 1 by the irradiation of the excitation light from the light propagation probe 5 enters from the glass sphere portion 6 in contact with the living tissue 1, and is spectrophotometrically measured through the optical fiber portion 7 and the fluorescence propagation optical system 62. It is incident on the means 10. Spectrophotometer 10
At this time, the light incident from the glass sphere portion 6 is spectrophotometrically measured to obtain light spectrum intensity distribution data.

Similarly to the above, when the living tissue 1 is irradiated with the excitation light, the light incident through the light propagation probe is spectrophotometrically measured by the spectrophotometric means 10 a plurality of times to obtain a plurality of optical spectrum intensity distribution data. get.

The plurality of optical spectrum intensity distribution data are output from the spectrophotometer 10 to the variation calculator 15, and the variation calculator 15 calculates the plurality of optical spectrum intensity distribution data from the values of the plurality of optical spectrum intensity distribution data. Variation is required.

The variation calculated by the variation calculating means 15 is output to the contact determining means 20, and the contact determining means 20 determines whether or not the light propagation probe 5 is in contact with the living tissue 1 based on this variation. . Further, a profile of a standardized spectrum intensity distribution, which will be described later, obtained from the variation calculation means 15 on the basis of the plurality of optical spectrum intensity distribution data is displayed on the display means 3.
5, and the profile is displayed on the display means 35.

Here, acquisition of the optical spectrum intensity distribution data by the spectrophotometric means 10 and variation calculation means 15
The calculation of the variation in and the above determination by the contact determination means 20 based on this variation will be described in detail.

As shown in FIG. 2, when the light propagation probe 5 comes into contact with the living tissue 1 in a predetermined contact state and the positional relationship between the living tissue and the light propagation probe is stable, the spectrophotometric light propagates. Since there is almost no mucus 2 covering the living tissue 1 in the region E sandwiched between the living tissue 1 and the glass sphere portion 6 in the optical path to be formed, at this time, the light spectrophotometrically measured by the spectrophotometric means 10 a plurality of times Tissue fluorescence occupies most of the amount of light, and the acquired plurality of light spectrum intensity distribution data are the tissue fluorescence spectrum intensity distribution data with a small variation and a high S / N. The normalized profile of the spectrum intensity distribution created by inputting these optical spectrum intensity distribution data to the variation calculation means 15 is created with the ordinate as the normalized intensity and the abscissa as the measurement wavelength.
There is little variation in shape, as indicated by P1, P2, and P3 inside. In addition, to normalize means to represent the intensity of fluorescence at a specific measurement wavelength spectrophotometrically represented by the ratio of the value of this intensity and the value obtained by integrating the intensity of the fluorescence over the entire measurement wavelength region. The area S surrounded by the profile of the spectrum intensity distribution standardized in this way becomes equal for all the acquired light spectrum intensity distribution data. The mucus 2 is covered with air 3.

On the other hand, when the light propagation probe 5 is not in contact with the living tissue 1 in a predetermined contact state and the positional relation between the living tissue 1 and the light propagation probe 5 is not stable, the As shown in FIG. 4, the state where the glass sphere 6 does not contact the living tissue 1 or the state where the area where the glass sphere 6 contacts the living tissue 1 increases or decreases Repeatedly, the mucus 2 enters the area E sandwiched between the living tissue 1 and the glass bulb 6,
Moreover, since the range of this region E changes, the ratio of the respective amounts of tissue fluorescence and mucus fluorescence incident on the light propagation probe and spectrophotometrically changed, and at this time, a plurality of light spectrophotometrically measured by the spectrophotometric means 10 is changed. The spectral intensity distribution data will be different each time spectrophotometry is performed, and the variation will be large.
Therefore, the standardized profile of the spectrum intensity distribution created by inputting these optical spectrum intensity distribution data to the variation calculating means 15 is P4 in FIG.
As shown in P5 and P6, the variation in shape becomes large.

Next, FIG. 3 showing the normalized profiles P1, P2, P3 for the calculation of the variation in the variation calculation means 15 and the above determination by the contact determination means 20, and the above-mentioned normalized profile. It demonstrates using FIG. 5 in which P4, P5, and P6 are shown.

The first variation calculation method calculates the variation based on the difference between a plurality of normalized profiles of the spectrum intensity distribution, and has a specific wavelength, for example, wavelength 450 nm, wavelength 550 nm, wavelength 65.
The variation calculation means 15 calculates the variation widths W1, W2, W3 of the plurality of profiles at 0 nm.

Then, when all of the widths W1, W2, W3 of the variation by the contact determination means 20 are less than a predetermined width R1 (W1≤R1, and W2≤R1,
When W3 ≦ R1), it is determined that the light propagation probe is in contact with the biological tissue, and any one of W1, W2, and W3 is larger than a predetermined width R1 (W
1> R1 or W2> R1 or W3> R1), it is determined that the light propagation probe is not in contact with the biological tissue.

Therefore, as shown in FIG. 3, P1, P
In the measurement in which the data of 2 and P3 were acquired, it was determined that the light propagation probe was in contact with the biological tissue (W1 ≦ R
1, and W2 ≦ R1 and W3 ≦ R1), and in the measurement in which the data of P4, P5, and P6 are acquired as shown in FIG. 5, it is determined that the light propagation probe is not in contact with the biological tissue (W2 > R1, W3> R1).

The second variation calculation method is to calculate the variation based on the difference between two consecutive data among the plurality of optical spectrum intensity distribution data. For example, assuming that the data of P4, P5, and P6 shown in FIG. 5 are acquired in this order, they are continuously acquired at the same specific wavelength as the above, for example, wavelength 450 nm, wavelength 550 nm, and wavelength 650 nm. P4, P5
Between the data differences V1, V2, V3 (see FIG. 6) and P
5 and P6 data difference V1 ', V2', V3 '(see FIG. 7).
And (see reference) are calculated by the variation calculation means 15.

Then, by the contact determination means 20, V1,
When all of V2, V3, V1 ', V2', and V3 'are less than or equal to a predetermined width R2, it is determined that the light propagation probe is in contact with the living tissue, and V1, V2, V3, V
When any one of 1 ', V2', and V3 'is larger than the predetermined width R2, it is determined that the light propagation probe is not in contact with the living tissue.

When the contact determining means 20 determines that the light propagation probe 5 is not in contact with the living tissue 1, the contact determining means 20 outputs this determination result to the notifying means 25 and the lamp 26 included in the notifying means 25. Is lit.
On the other hand, when the contact determination means 20 determines that the light propagation probe 5 is in contact with the biological tissue 1, the contact determination means 20 outputs this determination result to the averaging means 30, and inputs this determination result. The averaging means 30 inputs the plurality of acquired optical spectrum intensity distribution data from the variation calculating means 15 and adds and averages them to calculate the tissue fluorescence spectrum intensity distribution data. The averaging means 30 further outputs the profile of the tissue fluorescence spectrum intensity distribution obtained on the basis of the tissue fluorescence spectrum intensity distribution data to the display means 35, and the display means 35 displays this profile.

When the excitation light is intermittently emitted from the LD light source 61 in the presence of background light, the spectrophotometric means 10
According to the above, a plurality of optical spectrum intensity distribution data may be acquired as follows. That is, from the LD light source 61 following or before the excitation light is irradiated from the light spectrum intensity distribution data obtained by spectrophotometrically measuring the light incident through the light propagation probe 5 while the excitation light is being irradiated. A plurality of data used for calculating the above-mentioned variation by subtracting background light spectrum intensity distribution data representing the background light spectrum intensity distribution obtained by spectrophotometrically measuring the light incident through the light propagation probe 5 when the excitation light is not irradiated. You may make it acquire the optical spectrum intensity distribution data of.

The timing of the above operation is controlled by the controller (not shown). Further, the spectrophotometer 10 can be configured using, for example, a prism and a linear CCD.

FIG. 8 is a block diagram showing the schematic arrangement of a fluorescence spectrum acquisition apparatus according to the second embodiment of the present invention, and FIG.
FIG. 10 is a diagram showing a first variation calculation method in the second embodiment, and FIG. 11 is a diagram showing a second variation calculation method in the second embodiment. The fluorescence spectrum acquisition apparatus of the second embodiment measures the intensity of light incident through a light propagation probe instead of the variation calculation means 15 and the contact determination means 20 of the fluorescence spectrum acquisition apparatus of the first embodiment. , A background light intensity photometric means 40 having a function of performing calculation and determination, a variation calculation means 45, and a contact determination means 50 are arranged. Others are the same as those of the first embodiment. Become. Hereinafter, the same components as those in the first embodiment will be designated by the same reference numerals and the description thereof will be omitted.

The fluorescence spectrum acquisition apparatus 102 of the second embodiment includes a light propagation probe 5 which makes fluorescence emitted from the living tissue 1 enter by irradiation of the excitation light emitted from the LD light source 61, and the light propagation probe 5. Spectrophotometric means 10 for spectrophotometrically measuring the fluorescence incident through the above to obtain the spectral intensity distribution data of the fluorescence, and the light propagation probe 5.
Fluorescence propagation optical system 62 for making the above-mentioned fluorescence incident through the above into the spectrophotometric means 10, and this fluorescence propagation optical system 62.
And an excitation light propagation optical system 63 for making the excitation light incident on the light propagation probe 5 from the side where the fluorescence incident on the light propagation probe 5 is emitted.

Further, this fluorescence spectrum acquisition apparatus 10
Reference numeral 2 denotes a white light source 81 that emits white light that is background light that illuminates the living tissue 1, the living tissue 1 is illuminated by this background light (white light), and the living tissue 1 is not irradiated with excitation light. The background light intensity photometric means 4 which obtains a plurality of background light intensity data representing the intensity of the background light by photometrically measuring the background light that is incident through the light propagation probe 5 a plurality of times.
0, a variation calculation unit 45 that calculates variations of the plurality of background light intensity data based on the plurality of background light intensity data, and the light propagation probe 5 uses the biological tissue 5 based on the variations calculated by the variation calculation unit 45. The contact determination means 50 for determining whether or not it is in contact with 1.

The background light intensity photometric means 40 is disposed between the third convex lens 76 and the spectrophotometric means 10, and the third light intensity photometric means 40 is provided.
The background light half mirror 41 reflects the light that is condensed by the convex lens 76 and goes toward the spectrophotometric means 10 and makes it enter the background light intensity photometric means 40.

Next, the operation of the above embodiment will be described.

After inserting the light propagation probe 5 into the living body and bringing the glass bulb portion 6 into contact with the living tissue 1, the white light source 81 is turned on to illuminate the living tissue 1 with white light.

When the living tissue 1 is illuminated by the white light and the exciting light is not applied to the living tissue, the white light incident through the light propagation probe 5 and the fluorescence propagation optical system 62 is measured as the background light intensity measuring means. The background light intensity photometric means 40 obtains a plurality of background light intensity data representing the intensity of the white light obtained by measuring the light multiple times by 40.

The plurality of background light intensity data obtained by the background light intensity photometry means 40 are input to the variation calculation means 45, and the variation calculation means 45 obtains the variations of the plurality of background light intensity data. Then, the contact determination means 50, which has received the variation from the variation calculation means 45, determines whether or not the light propagation probe 5 is in contact with the living tissue 1 based on this variation.

Now, the acquisition of the background light intensity data by the background light intensity photometry means 40, the calculation of the variation in the variation calculation means 45, and the above determination by the contact determination means 20 based on this variation will be described in detail.

When the light propagation probe 5 is brought into contact with the living tissue 1 in a predetermined contact state and the positional relationship between the living tissue 1 and the light propagation probe 5 is stable, as shown in FIG. Since there is almost no air 3 or mucus 2 covering the living tissue 1 in the area E sandwiched by the portion 6, the optical path through which the white light emitted from the white light source 81 enters the light propagation probe 5 is almost closed. At this time, the amount of light that is measured a plurality of times by the background light intensity photometric means 40 is extremely small and its variation is small. The intensity of the background light represented by the background light intensity data acquired by the background light intensity photometric means 40 is also G1, G2, G3, G4 in FIG. 9 in which the vertical axis is the light intensity and the horizontal axis is the measurement time. As shown by, there is little variation.

On the other hand, when the light propagation probe 5 is not in contact with the living tissue 1 in a predetermined contact state and the positional relationship between the living tissue 1 and the light propagation probe 5 is not stable, it is shown in FIG. As described above, the state in which the glass bulb portion 6 does not contact the living tissue 1 or the state in which the area of the region where the glass bulb portion 6 contacts the living tissue 1 increases or decreases is repeated, Air 3 and mucus 2 enter the area E sandwiched between the living tissue 1 and the glass bulb portion 6,
Moreover, since the range of the area E changes, the amount of the background light reflected by these boundary surfaces and incident on the light propagation probe also changes, and at this time, the background light measured by the background light intensity photometric means 40. The intensity data is different for each measurement, and the light intensity represented by this background light intensity data has large variations as shown by G5, G6, G7, and G8 in FIG.

Next, regarding the calculation of the variation in the variation calculation means 15 and the above determination by the contact determination means 20, FIG. 9 in which the intensity of the background light is shown by G1, G2, G3 and G4, and G5, G6 and G7. , G8, the intensity of the background light is shown in FIG.

The first variation calculation method is to calculate the variation based on the difference in light intensity.
Width W of variation in light intensity represented by 1, G2, G3, G4 (see FIG. 9), or G5, G6, G7, G
The width W (see FIG. 10) of the variation in the light intensity represented by 8 is calculated by the variation calculation means 45.

Then, the contact determining means 20 determines that the light propagation probe is in contact with the living tissue when the width W of the variation is less than or equal to the predetermined width L1, and the width W of the variation is determined in advance. Specified predetermined width L1
When it is larger, it is determined that the light propagation probe is not in contact with the living tissue.

Therefore, as shown in FIG. 9, G1, G
In the measurement in which the data of 2, G3, and G4 were acquired, it was determined that the light propagation probe was in contact with the biological tissue (W
≦ L1), G5, G6, G7, G8 as shown in FIG.
In the measurement in which the data of 1 is acquired, it is determined that the light propagation probe is not in contact with the biological tissue (W> L1).

The second variation calculation method is to calculate the variation based on the difference between two consecutive data among the plurality of background light intensity data. For example, as shown in FIG. 11, continuously acquired data G5, G
Difference V1 between 6 and data G6 and G7 continuously acquired
The difference calculation unit 45 calculates a difference V2 between them and a difference V3 between the continuously acquired data G7 and G8.

Then, the contact determination means 50 causes V1,
When all of V2 and V3 are less than or equal to a predetermined width L2 that is predetermined, it is determined that the light propagation probe is in contact with the biological tissue, and any one of V1, V2, and V3 is a predetermined width that is predetermined. When it is larger than L2, it is determined that the light propagation probe is not in contact with the living tissue.

When the contact determination means 50 determines that the light propagation probe 5 is not in contact with the living tissue 1, the contact determination means 50 outputs this result to the notification means 25, and the lamp 26 included in the notification means 25 operates. It is lit. On the other hand, when the contact determination means 50 determines that the light propagation probe 5 is in contact with the biological tissue 1, the white light source 81 is turned off, the LD light source 61 is turned on, and the LD light source 6 is turned on.
The fluorescence generated from the living tissue 1 by the irradiation of the excitation light emitted from the device 1 is spectrophotometrically measured by the spectrophotometric device 10, and a plurality of optical spectrum intensity distribution data are acquired as in the first embodiment. Then, the plurality of light spectrum intensity distribution data are input to the averaging means 30 and added and averaged to obtain the tissue fluorescence spectrum intensity distribution data.
This tissue fluorescence spectrum intensity distribution data is displayed by the display means 35.
Will be output and displayed.

The timing of the above operation is controlled by a controller (not shown).

Further, the background light may include background light different from the white light emitted from the white light source.

Further, the fluorescence spectrum acquisition device may be configured so that the spectrophotometric means also serves as the background light photometric means, and the determination method by the contact determination means is not limited to the above method, and any method may be used. good.

Further, the notifying means for notifying the result of the determination is not limited to the lamp and may be one for notifying by sound or vibration, and the result of the determination may be displayed on the display means.

The light source for emitting the excitation light is not limited to the LD light source, and may be a light source using a gas laser, a mercury lamp, or the like, and the wavelength of the excitation light is not limited to 410 nm, and fluorescence is emitted from the living tissue. The light may have any wavelength as long as it can.

[Brief description of drawings]

FIG. 1 is a block diagram showing a schematic configuration of a fluorescence spectrum acquisition device according to a first embodiment.

FIG. 2 is a diagram showing a state in which a light propagation probe is in contact with living tissue in a predetermined contact state.

FIG. 3 is a diagram illustrating a first variation calculation method according to the first embodiment.

FIG. 4 is a diagram showing a state in which the light propagation probe is not in contact with living tissue in a predetermined contact state.

FIG. 5 is a diagram showing a first variation calculation method in the first embodiment.

FIG. 6 is a diagram showing a second variation calculation method in the first embodiment.

FIG. 7 is a diagram showing a second variation calculation method in the first embodiment.

FIG. 8 is a block diagram showing a schematic configuration of a fluorescence spectrum acquisition device according to a second embodiment.

FIG. 9 is a diagram showing a calculation method of a first variation in the second embodiment.

FIG. 10 is a diagram showing a calculation method of a first variation in the second embodiment.

FIG. 11 is a diagram illustrating a second variation calculation method according to the second embodiment.

[Explanation of symbols]

1 living tissue 5 Light propagation probe 6 glass bulbs 7 Optical fiber part 10 spectrophotometric means 10 15 Variation calculation means 20 Contact determination means 25 Notification means 30 means of averaging 35 display means 61 LD light source 62 Fluorescence propagation optical system 63 Excitation light propagation optical system 101 Fluorescence spectrum acquisition device

Continued front page    F-term (reference) 2G043 AA03 BA16 CA09 EA01 FA01                       GA06 GB01 GB21 HA01 HA02                       HA05 HA09 JA01 KA09 LA01                       MA01 NA01                 4C061 AA00 BB02 CC07 DD03 HH51                       JJ17 NN01 NN05 QQ04 SS18                       SS21 WW17

Claims (5)

[Claims] 1. A fluorescence spectrum in which fluorescence generated from a biological tissue by irradiation with excitation light is incident through a light propagation probe, and the fluorescence incident through the light propagation probe is spectrophotometrically measured to obtain spectrum intensity distribution data of the fluorescence. In the acquisition method, a plurality of light spectrum intensity distribution data is acquired by spectrophotometrically measuring the light incident through the light propagation probe when the excitation light is applied to the living tissue, and the plurality of light spectra. A fluorescence spectrum characterized by determining the variation of the plurality of optical spectrum intensity distribution data from the value of the intensity distribution data, and determining whether or not the light propagation probe is in contact with the biological tissue based on the variation. Acquisition method. 2. A fluorescence spectrum in which fluorescence generated from living tissue by irradiation with excitation light is made incident through a light propagation probe, and the fluorescence made incident through the light propagation probe is spectrophotometrically measured to obtain spectrum intensity distribution data of the fluorescence. In the acquisition method, the living tissue is illuminated with background light, and when the excitation light is not applied to the living tissue, the background light incident through the light propagation probe is measured a plurality of times to obtain the background light. Acquiring a plurality of background light intensity data representing the intensity of the, the variation of the plurality of background light intensity data from the value of the plurality of background light intensity data, the light propagation probe to the biological tissue based on the variation. A method for acquiring a fluorescence spectrum, which comprises determining whether or not they are in contact with each other. 3. A light-propagating probe that makes fluorescence emitted from living tissue upon irradiation with excitation light enter, and spectrophotometry that spectrophotometrically measures fluorescence that has entered through the light-propagating probe to obtain spectral intensity distribution data of the fluorescence. In the fluorescence spectrum acquisition device including means, when the excitation light is irradiated to the biological tissue, the spectrophotometric means, spectrophotometric multiple times the light incident through the light propagation probe a plurality of For obtaining the optical spectrum intensity distribution data, a variation calculation means for calculating the variation of the plurality of optical spectrum intensity distribution data based on the values of the plurality of optical spectrum intensity distribution data obtained by the spectrophotometric means, When the light propagation probe comes into contact with the biological tissue based on the variation calculated by the variation calculation means. Fluorescence spectrum acquisition apparatus characterized by comprising a contact determination means for determining whether Luke. 4. When the excitation light is intermittently irradiated in the presence of background light, the spectrophotometric means measures the light incident through the light propagation probe when the excitation light is irradiated. Background light obtained by spectrophotometrically measuring the background light incident through the light propagation probe when the excitation light is not irradiated following the irradiation of the excitation light from the light spectrum intensity distribution data obtained by spectrophotometry 4. The background light spectrum intensity distribution data representing the spectrum intensity distribution is subtracted to obtain the light spectrum intensity distribution data used for calculating the variation.
A fluorescence spectrum acquisition apparatus using the light propagation probe described. 5. A light-propagating probe that makes fluorescence emitted from living tissue upon irradiation with excitation light enter, and spectrophotometry that spectrophotometrically measures fluorescence that has entered through the light-propagating probe to obtain spectral intensity distribution data of the fluorescence. In the fluorescence spectrum acquisition device including means, the background light incident through the light propagation probe when the living tissue is illuminated by background light and the excitation light is not applied to the living tissue is measured a plurality of times. And a background light intensity metering unit for acquiring a plurality of background light intensity data representing the intensity of the background light, and a variation calculation unit for calculating a variation of the plurality of background light intensity data based on the plurality of background light intensity data. And whether or not the light propagation probe is in contact with the biological tissue based on the variation calculated by the variation calculation means. Fluorescence spectrum acquisition apparatus characterized by comprising a contact determination means.