JP2005049282A - Fluorescent observation apparatus and laser light irradiation device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a fluorescent observation apparatus capable of miniaturizing the whole device, facilitating handleability, and reducing photobreaching. <P>SOLUTION: This fluorescent observation apparatus is characterized by being equipped with an imaging device 1 for imaging an image by fluorescence from an observation object 6, an illumination device 2 having a stroboscopic light source 22 for emitting illumination light to the observation object 6, an excitation light generation filter 3 provided on the position on the observation object 6 side relative to the illumination device 2, for extracting light having a prescribed excitation wavelength from the illumination light from the illumination device 2, and generating excitation light for generating the fluorescence, and an excitation light removal filter 4 provided on the position on the observation object 6 side relative to the imaging device 1, for removing the light having the prescribed excitation wavelength. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a fluorescence observation apparatus and a laser beam irradiation apparatus.

  In recent years, photodynamic diagnosis of tumors has been made using the property that fluorescent substances are emitted when a photosensitive substance that is accumulated more in tumor cells than in normal cells is administered to a living body, and the photosensitive substance is excited by light of a specific wavelength. Photodynamic diagnosis (PDD) and photodynamic therapy of tumors (PDT: photodynamic diagnosis) utilizing the property of generating singlet oxygen and destroying tumor cells when a photosensitive substance is excited by light of a specific wavelength Photodynamic therapy) is drawing attention.

  Photosensitive substances used in PDD and PDT include, for example, 5-ALA (manufactured by Cosmo Research Institute), ATX-S10 (manufactured by Photochemical Research Institute), and the like. These are known to emit red fluorescence when excited with blue light. When a blue excitation light is irradiated to a diagnostic object to which these substances have been administered in advance, red fluorescence is generated more strongly from tumor cells that accumulate more of these substances than normal cells. Therefore, by observing this fluorescence, the tumor site in the living body can be specified.

  In general, in order to observe fluorescence, a fluorescence observation apparatus including a light source that emits excitation light applied to an observation target and an imaging unit that images fluorescence generated from the observation target is used. However, since the fluorescence emitted from the above-mentioned photosensitive substance is buried in the excitation light and is weak, it is difficult to observe.

  However, in recent years, with the emergence of high-sensitivity detectors capable of observing this fluorescence, research and development are being promoted toward specific diagnosis of tumor sites. For example, Non-Patent Document 1 describes a fluorescence observation apparatus aimed at finding skin cancer.

Note that prior art document information relating to the invention of this application includes the following.
Japanese Patent Laid-Open No. 2001-078205 Japanese Patent No. 3028421 JP 2001-299941 A Journal of Modern Optics, 2000 Vol.47, No.11, 2021-2027

  In a conventional fluorescence observation apparatus, a xenon lamp or the like is used as a light source, and a camera or the like equipped with a cooled CCD or an ICCD using an image intensifier is used as an imaging unit.

  However, with such a configuration, there is a problem that the entire apparatus is large and handling is not easy. For this reason, such an apparatus cannot be said to be sufficient in terms of applicability and practicality of the apparatus, such as that its use is mainly for research. In addition, when a normal xenon lamp or the like is used as an excitation light source, there has been a problem of photobleaching in which the fluorescence characteristics of the photosensitive material change when irradiated with excitation light for a long time.

  The present invention has been made in order to solve such problems, and the fluorescence observation apparatus and the fluorescence observation apparatus in which the apparatus is miniaturized, the handling is facilitated, and the photobleaching is reduced. An object of the present invention is to provide a laser beam irradiation apparatus using the above.

  In order to achieve such an object, a fluorescence observation apparatus according to the present invention includes an imaging unit that captures an image of fluorescence from an observation target, an illumination unit that includes a strobe light source that emits illumination light to the observation target, and an illumination. An excitation light generating means that is provided at a position on the observation object side with respect to the means, extracts light of a predetermined excitation wavelength from the illumination light from the illumination means, and generates excitation light for generating fluorescence; and imaging And a light removing unit that is provided at a position on the observation target side with respect to the unit and removes light having a predetermined excitation wavelength.

  Thus, in the fluorescence observation apparatus according to the present invention, the illuminating means having the strobe light source (flash lamp) provided together with the imaging means is used as the excitation light source. For this reason, compared with the case where the conventional xenon lamp is used etc., the whole apparatus is reduced in size and handling becomes easy. Furthermore, by using the flash light emitted from the strobe light source as the excitation light, the time for irradiating the excitation light can be shortened and the photobleaching of the photosensitive substance can be reduced. Further, by providing excitation light generation means and light removal means for these imaging means and illumination means, respectively, it becomes possible to make the observation device having the above configuration function well as a fluorescence observation device.

  In addition, it is preferable to use an imaging device in which the imaging unit and the illumination unit are integrally provided in advance as the imaging unit and the illumination unit.

  In this case, the entire apparatus can be further reduced in size as compared with the case where the imaging unit and the illumination unit are provided separately. For example, a commercially available camera with a strobe can be used as the imaging device.

  The excitation light generation means has a first filter that transmits light of a predetermined excitation wavelength and is detachable from the illumination means. The light removal means removes light of the predetermined excitation wavelength and serves as an imaging means. It is preferable to have the 2nd filter which can be attached or detached with respect to.

  In this case, a normal image can be acquired by capturing an image with the first filter and the second filter removed from the apparatus. That is, although there is only one apparatus, both normal light observation and fluorescence observation can be performed by a simple switching operation of attaching / detaching a filter.

  The fluorescence observation apparatus further includes a scale projection unit that projects a scale onto the observation target with illumination light from the illumination unit, and the imaging unit captures an image of fluorescence and an image of the scale projected onto the observation target. It is good also as a feature.

  In this case, since an image obtained by superimposing the fluorescence image and the scale image can be obtained, the actual size can be measured from the image, and the size of the specific part in the observation target can be known. .

  The laser light irradiation apparatus according to the present invention is provided on the optical path of the above-described fluorescence observation apparatus, a laser light source that emits laser light for irradiating the observation target, and the laser light that travels from the laser light source to the observation target. And a spatial light modulation means for spatially modulating the laser beam based on the information of the image of the fluorescence imaged in (1).

  Thus, in the laser beam irradiation apparatus according to the present invention, the laser beam generated from the laser light source is spatially modulated based on the information of the fluorescence image captured by the fluorescence observation apparatus before irradiating the observation target. Received from light modulation means. For this reason, it is possible to selectively irradiate a specific part in the observation target with the laser beam based on the fluorescence image.

  The fluorescence observation apparatus according to the present invention uses illumination means having a strobe light source as an excitation light source. For this reason, compared with the case where the conventional xenon lamp is used etc., the whole apparatus is reduced in size and handling becomes easy.

  Furthermore, by using the flash light emitted from the strobe light source as the excitation light, the time for irradiating the observation target with the excitation light is shortened. For this reason, when performing fluorescence observation using a photosensitive substance, photobleaching of the photosensitive substance can be reduced.

  The fluorescence observation apparatus according to the present invention can function well as a fluorescence observation apparatus by providing excitation light generation means and light removal means for the imaging means and illumination means, respectively.

  Hereinafter, preferred embodiments of a fluorescence observation apparatus and a laser beam irradiation apparatus according to the present invention will be described in detail with reference to the drawings. In the description of the drawings, the same elements are denoted by the same reference numerals, and redundant description is omitted. Further, the dimensional ratios in the drawings do not necessarily match those described.

(First embodiment of fluorescence observation apparatus)
FIG. 1 is a side view schematically showing a configuration of a first embodiment of a fluorescence observation apparatus according to the present invention. This fluorescence observation apparatus irradiates the observation object 6 with excitation light and images the fluorescence generated in the observation object 6 as a two-dimensional image. As shown in the figure, the fluorescence observation apparatus includes an imaging device 1, an illumination device 2, an excitation light generation filter (first filter) 3, and an excitation light removal filter (second filter) 4.

  The imaging device 1 is an imaging unit that captures an image (fluorescence image) of fluorescence from the observation target 6 with a predetermined imaging axis. The imaging device 1 includes an imaging element 11, a lens unit 7, a control unit 12, and an operation unit 14. The image sensor 11 is an element such as a CCD having pixels arranged two-dimensionally, and converts a received optical image into an electric signal to generate two-dimensional image data. A lens unit 7 is provided on the front surface of the imaging device 1 (a surface facing the observation object 6) with respect to the imaging element 11. The lens unit 7 includes an optical system that forms an image of incident light such as fluorescence from the observation target 6 onto the image sensor 11.

  The control unit 12 is a control unit that controls the imaging operation of the imaging apparatus 1. The control unit 12 is connected to the image sensor 11 and controls the operation of the image sensor 11. An operation unit 14 is connected to the control unit 12. The operation unit 14 is provided on the outer surface of the imaging apparatus 1 and can be operated by an observer. The operation unit 14 instructs the control unit 12 to execute imaging, or if necessary, instructs imaging conditions.

  In addition, the imaging device 1 includes an image recording unit 13 and an image display unit 8. The image recording unit 13 is connected to the image sensor 11 and records image data acquired by the image sensor 11 in a memory. The image display unit 8 is preferably a liquid crystal display, and is provided on the outer surface of the imaging device 1 in the same manner as the operation unit 14. The image display unit 8 displays the image of the observation target 6 captured by the image sensor 11 based on the image data recorded in the memory of the image recording unit 13. In addition, the imaging apparatus 1 is positioned and installed by a support member 9 such as a tripod so that its imaging axis faces the observation target 6.

  The illumination device 2 is an illumination unit that illuminates the observation target 6 with a predetermined illumination axis. The illumination device 2 includes a strobe light source 22 and a light source drive unit 21. The strobe light source 22 emits illumination light for the observation target 6. The light source driving unit 21 is connected to the strobe light source 22 and controls driving of the strobe light source 22. The light source driving unit 21 is connected to the control unit 12 of the imaging device 1 directly or via another control device or the like, and can drive and control the strobe light source 22 in accordance with the imaging timing of the imaging device 1. It is possible. The illuminating device 2 is attached to the imaging device 1 so that its illumination axis forms a predetermined angle with the imaging axis and faces the observation object 6.

  The excitation light generation filter 3 is excitation light generation means for extracting light with a predetermined excitation wavelength (light with a predetermined excitation wavelength band) from the illumination light from the strobe light source 22 of the illumination device 2. The excitation light generation filter 3 is provided at a position on the observation object 6 side with respect to the illumination device 2 so that light traveling from the strobe light source 22 toward the observation object 6 passes. As a result, excitation light for generating fluorescence is generated from the illumination light emitted from the strobe light source 22 and irradiated onto the observation object 6.

  The excitation light removal filter 4 is light removal means for removing light having a predetermined excitation wavelength (light having a wavelength band including at least the excitation wavelength band). Further, the excitation light removal filter 4 is provided at a position on the observation object 6 side with respect to the imaging apparatus 1 so that light traveling from the observation object 6 to the imaging apparatus 1 passes.

  The fluorescence observation apparatus in the present embodiment further includes a projection scale plate 5. The projection scale plate 5 is a scale projection unit that projects the scale onto the observation target 6 with the illumination light from the illumination device 2 and is used as necessary. In FIG. 1, only the optical path of illumination light branched to the excitation light generation filter 3 and the projection scale plate 5 is schematically shown. Specific examples of the configuration of these optical systems will be described later.

  Next, the operation of the fluorescence observation apparatus according to the present embodiment will be described. When the observer gives an imaging instruction to the operation unit 14, the operation unit 14 sends an imaging instruction signal to the control unit 12. Upon receiving the instruction signal, the control unit 12 operates the light source driving unit 21 and the image sensor 11 while synchronizing the emission of the illumination light from the strobe light source 22 and the image pickup by the image sensor 11.

  The strobe light source 22 driven by the light source driving unit 21 emits illumination light. This illumination light passes through the excitation light generation filter 3 to become excitation light, and is irradiated on the observation object 6.

  Fluorescence from the observation target 6 generated by the excitation light enters the image sensor 11 through the excitation light removal filter 4 and the lens unit 7. At this time, the excitation light removal filter 4 removes light having a predetermined excitation wavelength including excitation light reflected by the observation object 6. The image sensor 11 captures an optical image by fluorescence. The obtained image data is recorded in the memory in the image recording unit 13 and the captured image is displayed in the image display unit 8.

  As described above, in this embodiment, the strobe light source 22 is used as the excitation light source. For this reason, this fluorescence observation apparatus is reduced in size as a whole and easy to handle as compared with the conventional case where a xenon lamp is used as a light source. Furthermore, by using the flash light emitted from the strobe light source 22 as the excitation light, the time for irradiating the excitation light can be sufficiently shortened with a simple configuration.

  In such a configuration, the excitation light generation filter 3 is provided as excitation light generation means for the illumination device 2. Therefore, the strobe light source 22 that emits white light can be used as the excitation light source, and the observation object 6 can be irradiated with the excitation light with a simple configuration.

  Further, the imaging device 1 is provided with an excitation light removal filter 4 as light removal means. Thereby, excess light such as excitation light reflected by the observation object 6 can be effectively removed from the light incident on the imaging device 1 from the observation object 6. Such removal of light is particularly effective when weak fluorescence is buried in excitation light. That is, by providing the excitation light removal filter 4 as described above, it is possible to remove excitation light and reliably observe weak fluorescence.

  The excitation light generating means and the excitation light removing means are not limited to the transmission type optical filters 3 and 4 as used in this embodiment, and various kinds of them may be used. For example, a reflective optical filter may be used.

  Furthermore, in the present embodiment, the captured image is configured to be displayed on the image display unit 8 provided in the imaging device 1. For this reason, the captured image can be immediately displayed to the observer with a simple configuration. As such an image display part 8, it is preferable to use what can be installed on the outer surface of the imaging device 1, such as a liquid crystal display. Alternatively, a configuration in which a CRT display or the like is installed externally and connected to the imaging apparatus 1 may be employed. Alternatively, the configuration may be such that the image display unit 8 is not provided when unnecessary, such as when automatically analyzing a captured image.

  Such a fluorescence observation apparatus can be used for PDD for skin cancer diagnosis, for example. In PDD using a fluorescence observation apparatus, a living body to which a photosensitive substance has been administered in advance is used as an observation target 6, and fluorescence from a photosensitive substance (drug fluorescence) selectively accumulated in a tumor site and fluorescence from a normal site (in-house) An image by fluorescence is captured.

  In this case, it is possible to diagnose the presence / absence of a tumor and the position / size / range of the tumor site from the captured image. At this time, since autofluorescence from the normal site also appears in the image, the position and range of the tumor site in the living body can be easily identified.

  Further, flash light emitted from the strobe light source 22 is used as excitation light. For this reason, the time for which the observation target 6 is irradiated with the excitation light is shortened, and photobleaching of the photosensitive substance is reduced.

  FIG. 2 is a graph showing an example of an excitation light spectrum and a fluorescence spectrum when the fluorescence observation apparatus is used for PDD. In this example, it is assumed that ATX-S10 is used as a photosensitive substance. In the figure, the horizontal axis represents wavelength (nm) and the vertical axis represents the relative intensity of excitation light or fluorescence. The wavelength is in the visible light range of 400-700 nm.

  In this case, as the excitation light, light having a wavelength range of approximately 400 to 450 nm as indicated by the excitation light spectrum 31 in the figure can be used. In order to generate such excitation light, a band pass filter that transmits light of approximately 400 to 450 nm may be used as the excitation light generation filter 3.

  Autofluorescence and drug fluorescence are generated on the longer wavelength side of the excitation light spectrum 31 as indicated by autofluorescence spectrum 32 and drug fluorescence spectrum 33 in the figure. In this example, the autofluorescence spectrum 32 is generated in a wavelength range of approximately 500 to 700 nm. In addition, the drug fluorescence spectrum 33 is generated in a wavelength region near 660 nm.

  Further, as the excitation light removal filter 4, as shown by the transmittance spectrum 34 of the excitation light removal filter in the figure (the vertical axis of the graph is shown as the transmittance), a cut that transmits light having a wavelength of about 480 nm or more is transmitted. An off filter can be used. In this case, autofluorescence and drug fluorescence in the wavelength range of 480 nm or more pass through the excitation light removal filter 4. On the other hand, the excitation light in the wavelength range shorter than 480 nm does not pass through the excitation light removal filter 4. For this reason, the excitation light reflected by the observation object 6 out of the light from the observation object 6 is removed, and autofluorescence and drug fluorescence selectively enter the imaging device 1.

  In addition, as the imaging device 1 and the illumination device 2, an imaging device in which they are integrally provided in advance may be used. In this case, the entire apparatus can be further downsized, and the fluorescence observation apparatus can be configured more easily.

  As such an imaging apparatus, for example, a commercially available digital camera with a strobe can be used. In such a digital camera, a shutter button is provided as the operation unit 14, and when the observer presses the shutter button and issues an instruction for imaging, emission of illumination light and imaging are executed in synchronization.

  Moreover, it is preferable that the excitation light generation filter 3 and the excitation light removal filter 4 are detachably attached to the strobe light source 22 and the imaging device 1, respectively.

  In this case, if the strobe light source 22 is operated with the excitation light generation filter 3 removed, the observation object 6 can be irradiated with white light. For this reason, the strobe light source 22 can be used as an excitation light source and a white light source. Further, when the imaging element 11 is operated with the excitation light removal filter 4 removed, an image (normal image) by reflected light from the observation target 6 can be captured. For this reason, the observer can observe both the normal image and the fluorescent image only by performing a simple switching operation of attaching and detaching the filter.

  Furthermore, in the present embodiment, a projection scale plate 5 may be used as necessary. As the projection scale plate 5, for example, a transparent plastic plate or a glass plate on which scales of actual dimensions are written can be used. When the projection scale plate 5 is used, as schematically shown in FIG. 1, the illumination light from the strobe light source 22 is divided into an optical path incident on the excitation light filter 3 and an optical path incident on the projection scale 5 plate. Branch. Then, a scale image is projected onto the observation object 6 by the light transmitted through the projection scale plate 5.

  In this case, the imaging device 1 captures an image of the scale projected on the observation target 6 together with the fluorescent image. As a result, an image in which the fluorescent image and the scale image are superimposed can be observed. For this reason, the actual size can be measured from the image, and the size of the specific part of the observation target 6 can be known. For example, when this fluorescence observation apparatus is used for PDD, the size of a tumor site or the like can be measured.

  FIG. 3 is a diagram showing an example of an image that can be observed when the projection scale plate 5 is used. As shown in the figure, the fluorescence image 51 and the scale image 52 appear on one image. In FIG. 3A, a cross scale is projected, and in FIG. 3B, a square scale is projected.

(Second embodiment of fluorescence observation apparatus)
FIG. 4 is a configuration diagram of a second embodiment of the fluorescence observation apparatus according to the present invention. Here, a configuration including an optical system is shown. As shown in the figure, the fluorescence observation apparatus according to the present embodiment includes an imaging system 100 and an illumination system 200.

  The imaging system 100 includes the imaging device 1, the excitation light removal filter 4, and an optical system. The imaging system 100 is installed such that its imaging axis faces the observation target 6. The imaging device 1 includes an imaging element 11, a control unit 12, an image recording unit 13, and an image display unit 8. The configuration of each part is the same as that of the embodiment shown in FIG. In addition, an excitation light removal filter 4 is installed at a predetermined position on the observation object 6 side with respect to the imaging apparatus 1.

  Between the imaging device 1 and the excitation light removal filter 4, a light guide optical system that guides the light from the observation object 6 that has passed through the excitation light removal filter 4 to the imaging device 1 is disposed along the imaging axis. Yes. In FIG. 4, the light guide optical system includes an imaging optical lens L <b> 4 that forms an image of incident light such as fluorescence on the imaging element 11.

  The illumination system 200 includes the illumination device 2, the excitation light generation filter 3, and an optical system. The illumination system 200 is installed such that its illumination axis is at a predetermined angle with the imaging axis of the imaging system 100 and faces the observation target 6. The lighting device 2 includes a strobe light source 22 and a light source driving unit 21. The configuration of each part is the same as that of the embodiment shown in FIG. Further, the excitation light generation filter 3 is installed at a predetermined position on the observation object 6 side with respect to the illumination device 2.

  Between the illumination device 2 and the excitation light generation filter 3, a light guide optical system that guides light from the strobe light source 22 to the observation object 6 through the excitation light generation filter 3 is disposed along the illumination axis. Yes. In FIG. 4, the light guide optical system includes condensing lenses L1 and L2 for condensing incident light, and a parallel light lens L3 for converting incident light into a parallel light flux. The condensing lenses L1 and L2 and the parallel light lens L3 are arranged in this order from the strobe light source 22 toward the excitation light generation filter 3.

  Next, the operation of the fluorescence observation apparatus according to the present embodiment will be described. Note that the operations of the portions not specifically described are the same as those in the above embodiment. The illumination light from the strobe light source 22 is condensed by the condensing lenses L1 and L2, and converted into a parallel light beam by the parallel light lens L3, and then enters the excitation light generation filter 3. The excitation light generated by the excitation light generation filter 3 is incident on the observation object 6.

  The fluorescence from the observation object 6 enters the imaging optical lens L4 through the excitation light removal filter 4. The imaging optical lens L4 condenses fluorescence so as to form an image on the imaging element 11. This fluorescent image is captured and displayed by the imaging device 1.

  As described above, in this embodiment, the strobe light source 22 is used as the excitation light source. For this reason, the fluorescence observation apparatus according to the present embodiment is downsized as a whole and is easy to handle.

  Furthermore, in this embodiment, no scale projection means is provided. For this reason, the fluorescence observation apparatus according to the present embodiment enables the fluorescence observation with a simple configuration when the scale projection means is not required.

(Third embodiment of fluorescence observation apparatus)
FIG. 5 is a configuration diagram of a third embodiment of the fluorescence observation apparatus according to the present invention. As shown in the figure, the fluorescence observation apparatus according to the present embodiment includes an imaging system 100, an illumination system 210, and a scale projection system 300. Among these, the configuration of the imaging system 100 is the same as that of the embodiment shown in FIG.

  The illumination system 210 includes an illumination system 2, an excitation light generation filter 3, and an optical system having lenses L1 to L3, similarly to the illumination system 200 in FIG. Furthermore, the illumination system 210 includes a half mirror HM1 that branches incident light into reflected light and transmitted light. The half mirror HM1 is disposed between the parallel light lens L3 and the excitation light generation filter 3 so as to be inclined at a predetermined angle with respect to the illumination axis. The light transmitted through the half mirror HM1 is irradiated to the observation object 6 as excitation light through the excitation light generation filter 3. On the other hand, the light reflected by the half mirror HM1 enters the scale projection system 300.

  The scale projection system 300 includes the projection scale plate 5 and an optical system. The scale projection system 300 is installed so that the projection axis thereof faces the observation target 6 so as to form a predetermined angle with respect to both the imaging axis of the imaging system 100 and the illumination axis of the illumination system 210. .

  The scale projection system 300 is provided with a mirror M1 for guiding the illumination light from the strobe light source 22 to the observation object 6 via the projection scale plate 5. The mirror M1 reflects the illumination light reflected by the half mirror HM1 of the illumination system 210, and changes its optical path in a direction along the projection axis. Further, a condensing lens L5 and a parallel light lens L6 are arranged in this order along the projection axis toward the observation target 6 between the projection scale plate 5 and the observation target 6. Further, the projection scale plate 5 is disposed at the focal position of the condensing lens L5.

  Next, the operation of the fluorescence observation apparatus according to the present embodiment will be described. Note that the operations of the portions not specifically described are the same as those in the above embodiment. The light from the strobe light source 22 enters the half mirror HM1 through the condensing lenses L1 and L2 and the parallel light lens L3. The half mirror HM1 splits the illumination light into excitation light generation transmitted light incident on the excitation light generation filter 3 and scale projection reflection light incident on the projection scale plate 5.

  The light for generating the excitation light is incident on the excitation light generation filter 3. The excitation light generated by the excitation light generation filter 3 is incident on the observation object 6.

  On the other hand, the light for scale projection is incident on the projection scale plate 5 after the optical path is changed by the mirror M1. The light for scale projection transmitted through the projection scale plate 5 is condensed by the condensing lens L5, converted into a parallel light beam by the parallel light lens L6, and then incident on the observation object 6.

  At this time, in the imaging system 100, the scale image projected onto the observation object 6 is captured and displayed together with the fluorescent image.

  As described above, in this embodiment, the strobe light source 22 is used as the excitation light source. For this reason, the fluorescence observation apparatus according to the present embodiment is downsized as a whole and is easy to handle.

  Further, in the present embodiment, a projection scale plate 5 is provided. For this reason, the actual size can be measured from the captured image, and the size of the specific part of the observation target 6 can be known.

(Fourth embodiment of fluorescence observation apparatus)
FIG. 6 is a configuration diagram of a fourth embodiment of the fluorescence observation apparatus according to the present invention. As shown in the figure, the fluorescence observation apparatus according to the present embodiment includes an imaging system 100, an illumination system 220, a scale projection system 310, and an optical axis adjustment system 500. Among these, the configuration of the imaging system 100 is the same as that of the embodiment shown in FIG.

  The illumination system 220 includes an illumination system 2, an excitation light generation filter 3, and an optical system having lenses L1 to L3, similarly to the illumination system 200 in FIG. Furthermore, the illumination system 220 includes a half mirror HM2 that branches incident light into reflected light and transmitted light. The half mirror HM2 is disposed on the downstream side of the parallel light lens L3 so as to be inclined with respect to the optical axis. The light reflected by the half mirror HM2 enters the half mirror HM3 provided in the optical axis adjustment system 500 as excitation light via the excitation light generation filter 3. On the other hand, the light transmitted through the half mirror HM2 enters the mirror M2 provided in the scale projection system 310.

  The scale projection system 310 includes the projection scale plate 5 and an optical system. The scale projection system 310 is provided with a mirror M 2 for guiding illumination light from the strobe light source 22 to the projection scale plate 5. The mirror M2 reflects the illumination light transmitted through the half mirror HM2 of the illumination system 220, and changes its optical path in a direction along a predetermined axis. A condensing lens L7 that condenses light for scale projection from the projection scale plate 5 is provided on the downstream side of the projection scale plate 5. The light that has passed through the projection scale plate 5 and the condensing lens L7 enters the half mirror HM4 of the optical axis adjustment system 500.

  The optical axis adjustment system 500 includes an objective lens L8 and half mirrors HM3 and HM4. The optical axis adjustment system 500 is installed so that its optical axis coincides with the imaging axis of the imaging system 100. The objective lens L8 is provided at a predetermined position facing the observation target 6, condenses the excitation light from the illumination system 220 and the light for scale projection from the scale projection system 310 on the observation target 6, and The fluorescence from the observation object 6 is guided to the imaging system 100.

  The half mirror HM3 is disposed between the objective lens L8 and the imaging system 100. The half mirror HM3 reflects the excitation light from the illumination system 220 and transmits the fluorescence from the observation target 6 and the like. The half mirror HM4 is disposed between the objective lens L8 and the half mirror HM3. The half mirror HM4 reflects the light for scale projection from the scale projection system 310 and transmits the fluorescence from the observation target 6 or the like. Further, the angles of the half mirror HM3 and the half mirror HM4 are adjusted so that the excitation light reflected by the half mirror HM4 and the light for scale projection are along the imaging axis of the imaging system 100.

  Next, the operation of the fluorescence observation apparatus according to the present embodiment will be described. Note that the operations of the portions not specifically described are the same as those in the above embodiment. Illumination light from the strobe light source 22 is split into excitation light generation light and scale projection light by the half mirror HM2. The excitation light emitted from the excitation light generation filter 3 and the scale projection light emitted from the projection scale plate 5 are incident on the half mirror HM3 and the half mirror HM4, respectively. The excitation light reflected by the half mirror HM3 and the light for scale projection reflected by the half mirror 4 enter the observation object 6 via the objective lens L8. At this time, the optical path through which these lights pass is along the imaging axis of the imaging system 100.

  The fluorescence from the observation object 6 and the light for scale projection reflected by the observation object 6 enter the imaging system 100 via the objective lens L8 and the half mirrors HM4 and HM3. In the imaging system 100, a fluorescent image and a scale image are captured and displayed.

  As described above, in this embodiment, the strobe light source 22 is used as the excitation light source. For this reason, the fluorescence observation apparatus according to the present embodiment is downsized as a whole and is easy to handle.

  Furthermore, in this embodiment, the light for scale projection from the projection scale plate 5 enters the observation target 6 through the optical path along the imaging axis of the imaging system 100 by the optical axis adjustment system 500. For this reason, it is possible to capture a scale image that does not include a dimensional error such as distortion. In addition, the size of the specific part of the observation target 6 can be obtained with high accuracy.

(Fifth embodiment of fluorescence observation apparatus)
FIG. 7 is a diagram showing a part of the configuration of the fifth embodiment of the fluorescence observation apparatus according to the present invention. Here, only the configuration of the optical system or the like in the case where the projection scale plate 5 is used is shown, and the other portions are not shown.

  In the present embodiment, as shown in the figure, a half mirror HM5, an excitation light generation filter 3, and a half mirror HM6 are provided between the illumination device 2 and the observation target 6. The illumination light from the illuminating device 2 that has passed through the half mirror HM5 passes through the excitation light generation filter 3 to become excitation light, and further passes through the half mirror HM6 and is irradiated onto the observation object 6.

  On the other hand, the light reflected by the half mirror HM5 is guided to the projection scale plate 5. Between the half mirror HM5 and the projection scale plate 5, a mirror M3 that reflects the illumination light from the half mirror HM5 and guides it to the projection scale plate 5 is provided. Further, a condensing lens L9 and a parallel light lens L10 are provided between the half mirror HM5 and the mirror M3. The illumination light from the half mirror HM5 is condensed by the condensing lens L9, converted into a parallel light beam by the parallel light lens L10, reflected by the mirror M3, and then incident on the projection scale plate 5.

  Further, a mirror M4 that reflects light from the projection scale plate 5 and guides it to the half mirror HM6 is provided between the projection scale plate 5 and the half mirror HM6. The light for scale projection that has passed through the projection scale plate 5 is reflected by the mirror M4 and then enters the half mirror HM6. The scale projection light is reflected by the half mirror HM6 and then enters the observation object 6.

(First embodiment of laser beam irradiation apparatus)
FIG. 8 is a plan view schematically showing the configuration of the first embodiment of the laser beam irradiation apparatus according to the present invention. This laser light irradiation apparatus selectively irradiates a specific part of the observation target 6 with laser light based on information of a fluorescent image captured by the fluorescence observation apparatus 10. As shown in the figure, the laser beam irradiation apparatus includes a fluorescence observation apparatus 10, an irradiation laser light source 41, and a spatial light modulation module 42. Among these, the fluorescence observation apparatus 10 captures / displays a fluorescence image or the like from the observation object 6, and for example, the one having the configuration shown in FIG. 1 is used.

  The irradiation laser light source 41 emits laser light for irradiating the observation target 6. The irradiation laser light source 41 is provided at a position where the laser light enters the observation object 6 via the spatial light modulation module 42.

  The spatial light modulation module 42 is a spatial light modulation unit that spatially modulates the laser light incident from the irradiation laser light source 41 based on information of the fluorescent image captured by the fluorescence observation apparatus 10. The spatial light modulation module 42 is provided on the optical path of the laser light that travels from the irradiation laser light source 41 to the observation target 6 so that the emitted laser light is incident on the observation target 6. In FIG. 8, a reflective spatial light modulation module 42 is used.

  Next, the operation of the laser beam irradiation apparatus according to the present embodiment will be described. The laser light irradiation apparatus includes (1) imaging of a fluorescent image by the fluorescence observation apparatus 10, (2) storage of a spatial modulation pattern by the spatial light modulation module 42, and (3) irradiation of laser light by the irradiation laser light source 42. Works in order. The operation of the fluorescence observation apparatus 10 is as described above.

  First, the fluorescence observation apparatus 10 captures a fluorescence image of the observation target 6. The spatial light modulation module 42 reads the information of the fluorescent image recorded in the memory and stores the spatial modulation pattern based on this information. Details of the operation of the spatial light modulation module 42 will be described later. Thereafter, when the operator gives an instruction, the irradiation laser light source 41 emits irradiation laser light. This irradiation laser beam is incident on the spatial light modulation module 42. The spatial light modulation module 42 spatially modulates the laser light with the stored spatial modulation pattern. The spatially modulated laser light is emitted from the spatial light modulation module 42 and applied to the observation object 6.

  As described above, in the present embodiment, the observation target 6 is irradiated with the laser light that has been spatially modulated in a predetermined pattern based on the fluorescence image. For this reason, it is possible to selectively irradiate the specific part of the observation target 6 with the laser beam based on the fluorescence image captured by the fluorescence observation apparatus 10. For example, when the laser beam irradiation apparatus according to the present embodiment is used for PDT treatment, it is possible to selectively irradiate the tumor site with the laser beam for treatment.

  As the spatial modulation pattern stored in the spatial light modulation module 42, the fluorescence pattern acquired by the fluorescence observation apparatus 10 may be used as it is. Alternatively, a computer for image processing may be further provided for the fluorescence observation apparatus 10. In this case, based on the information of the fluorescence image captured by the fluorescence observation device 10, a spatial modulation pattern is generated in this computer and sent to the spatial light modulation module 42.

  In such a configuration, it is possible to irradiate the laser beam after specifying the irradiation pattern in more detail. For example, by performing a process of deleting other pixel information while leaving a pixel that exhibits fluorescence of a predetermined intensity or higher, only a portion that emits fluorescence of a predetermined intensity or higher is irradiated with laser light. be able to. Alternatively, by performing processing such as deleting the other pixel information while leaving the pixel in which the fluorescence in the predetermined wavelength region appears, the laser light is irradiated only to the portion emitting the fluorescence in the predetermined wavelength region. You can also. In addition, the intensity of the laser beam applied to each part of the observation target 6 (the part corresponding to each pixel of the image to be captured) can be set to a constant value, and the intensity and wavelength of the fluorescence generated for each part. It can be a different value depending on.

  FIG. 9 is a diagram illustrating a configuration example of the spatial light modulation module 42. As shown in the figure, the spatial light modulation module 42 includes a spatial light modulator (SLM) 61, a writing light source 62, and a liquid crystal display (LCD) 63.

  The SLM 61 is a reflection type optical address type spatial light modulator, and is formed by having a transparent substrate 61A on the writing light incident side and a transparent substrate 61G on the reading light incident side. Between the transparent substrates 61A and 61G, in order from the transparent substrate 61A to the transparent substrate 61G, a transparent electrode 61B, a photoconductive layer 61C as an address layer, a dielectric mirror 61D, a liquid crystal layer 61E as a light modulation layer, and a transparent substrate An electrode 61F is formed.

  As the transparent substrate 61A constituting the SLM 61, for example, a glass substrate can be used. Alternatively, an optical image transmission element such as a fiber optical block may be used as the transparent substrate 61A. Further, when the distance between the LCD 63 and the optical image transmission element is set appropriately, the pixel structure of the LCD 63 is removed. In addition, as the transparent electrodes 61B and 61F, for example, ITO is used.

  At a position facing the transparent substrate 61A of the SLM 61, a writing light source 62 such as a semiconductor laser is disposed. The writing light source 62 emits light for writing a necessary spatial modulation pattern in the SLM 61.

  An LCD 63 is disposed between the SLM 61 and the writing light source 62. The LCD 63 is an electrical address type LCD for generating a predetermined pattern of writing light by the light from the writing light source 62. The LCD 63 includes a transparent substrate 63A on the writing light incident side and a transparent substrate 63E on the side facing the SLM 61. Between the transparent substrates 63A and 63E, an addressing element 63B, a liquid crystal layer 63C, and a transparent electrode 63D are formed in order from the transparent substrate 63A to the transparent substrate 63E. Note that the positional relationship between the addressing element 63B and the transparent electrode 63D may be reversed.

  In the above configuration, the spatial modulation pattern data from the fluorescence observation apparatus 10 is input to the LCD 63 by a VGA signal or the like, and a corresponding image is displayed on the LCD 63. Then, by passing the light from the light source 62 for writing through the LCD 63, writing light having a predetermined spatial modulation pattern is generated. Furthermore, when this writing light is incident on the SLM 61 from the transparent substrate 61A side, the impedance at each position of the photoconductive layer 61C changes according to the spatial modulation pattern. At this time, the voltage value applied to each position of the liquid crystal layer 61E changes, the alignment state of the liquid crystal changes, and the spatial modulation pattern is stored in the SLM 61.

  The SLM 61 in which the spatial modulation pattern is stored is irradiated with laser light from the irradiation laser light source 41 (see FIG. 8) as readout light from the transparent substrate 61G side. The laser light is reflected from the dielectric mirror 61D and then emitted from the SLM 61. At this time, the laser light undergoes spatial luminance modulation according to the alignment state of the liquid crystal layer 61E that passes therethrough. As a result, laser light spatially modulated into a predetermined pattern is emitted from the SLM 61. As the spatial light modulation means, various configurations other than those shown in FIG. 9 can be used.

  FIG. 10A is a diagram illustrating a configuration example of a laser beam irradiation apparatus. Here, the configuration of the spatial light modulation module 42 and the optical system for irradiating the observation target 6 with spatially modulated laser light is shown, and the fluorescence observation apparatus 10 is not shown.

  The spatial light modulation module 42 includes an SLM 61, a writing light source 62, and an LCD 63, similar to that shown in FIG. 9. Further, in FIG. 10A, a collimating optical system L11 that guides the writing light from the writing light source 62 to the LCD 63 as a parallel light beam is provided. The collimating optical system L11 is abbreviated as a single lens in the drawing, but actually comprises a lens group for correcting aberrations and the like. In this configuration example, the SLM 61 functions as an intensity modulation element.

  In FIG. 10A, an example is shown in which an image 71 (a hatched portion represents a dark portion of the image) in which a triangle is displayed at the center is input to the spatial light modulation module 42 as spatial modulation pattern data. It is shown as When the spatial light modulation module 42 in which the spatial modulation pattern based on the image 71 is stored is irradiated with laser light from the irradiation laser light source 41, the intensity of the laser light is modulated and reflected by the spatial light modulation module 42. The intensity-modulated laser light is imaged on the observation object 6 by the imaging optical system L12 provided in the optical path. As a result, an irradiation light image 72 having an intensity of 0 in other regions is formed on the observation object 6 while leaving a triangular pattern at the center. At this time, the spatial light modulation module 42 functions as a variable mask processing unit when irradiating the observation target 6 with laser light.

  FIG. 10B is a diagram illustrating another configuration example of the laser beam irradiation apparatus. Here, the configurations of the spatial light modulation module 42 and the optical system are the same as those in FIG. However, in this configuration example, the SLM 61 functions as a phase modulation element.

  FIG. 10B shows an example in which an image 73 having a square at the center is used as the spatial modulation pattern data. Here, the hologram 74 is synthesized from the image 73 of the target spatial modulation pattern using a computer, for example, by the simulated annealing method. Then, this hologram 74 is written into the spatial light modulation module 42. At this time, when the SLM 61 is irradiated with laser light, the SLM 61 performs phase modulation according to the hologram 74 on the laser light. On the observation object 6, an irradiation light image 75 in which the intensity of other regions is 0 is formed while leaving a square pattern at the center.

  In this case, since the laser light from the irradiation laser light source 41 is phase-modulated, all the light amounts of the laser light can be concentrated on the target pattern. For this reason, a laser beam can be used effectively.

  FIG. 11A is a diagram illustrating another configuration example of the laser light irradiation apparatus. Here, the configuration of the spatial light modulation module 42 is the same as that in FIG. In this configuration example, as shown in the figure, an imaging optical system L13 and a cubic half mirror 44 are used as an optical system for guiding laser light. The cubic half mirror 44 reflects the laser light from the irradiation laser light source 41 and guides it to the SLM 61 and transmits the laser light reflected by the SLM 61 to the observation target 6 via the imaging optical system L13. Lead. Further, the cubic half mirror 44 is arranged with an angle adjusted so that the laser light is incident on the SLM 61 substantially perpendicularly.

  Laser light from the irradiation laser light source 41 enters the SLM 61 via the cubic half mirror 44. The laser beam spatially modulated and reflected by the SLM 61 forms an image on the observation target 6 via the cubic half mirror 44 and the imaging optical system L13.

  In this case, since the laser beam is configured to enter the SLM 61 substantially perpendicularly, a uniform light output can be obtained.

  FIG. 11B is a diagram illustrating another configuration example of the laser beam irradiation apparatus. Here, the configurations of the spatial modulation module 42 and the optical system are the same as those in FIG. However, in this configuration example, the SLM 61 functions as a phase modulation element.

  In this configuration, similarly to FIG. 10B, the hologram is written in the spatial modulation module 42, and the spatial modulation module 42 performs spatial modulation by phase modulation on the laser light. For this reason, a laser beam can be used effectively. Further, as in FIG. 11A, a uniform light output can be obtained.

(Second Embodiment of Laser Light Irradiation Device)
FIG. 12 is a configuration diagram of a second embodiment of a laser beam irradiation apparatus according to the present invention. As shown in the drawing, the laser beam irradiation apparatus in this embodiment includes an imaging system 100, an illumination system 230, a scale projection system 320, a laser beam irradiation system 400, and an optical axis adjustment system 500. Among these, the imaging system 100, the illumination system 230, the scale projection system 320, and the optical axis adjustment system 500 constitute the fluorescence observation apparatus 10. Here, the positional relationship among the imaging system 100, the illumination system 230, the scale projection system 320, and the optical axis adjustment system 500 is the same as the positional relationship illustrated in FIG. 6 with respect to the fluorescence observation apparatus. The configurations of the imaging system 100 and the optical axis adjustment system 500 are also the same as those of the embodiment shown in FIG.

  The illumination system 230 includes an illumination system 2, an excitation light generation filter 3, a half mirror HM2, and an optical system having lenses L1 to L3, similarly to the illumination system 220 in FIG. Further, the illumination system 230 is provided with a half mirror HM7 on the downstream side of the excitation light generation filter 3. The half mirror HM7 reflects the laser light from the laser light irradiation system 400, guides it to the half mirror HM3 of the optical axis adjustment system 500, and transmits the excitation light from the excitation light generation filter 3. A condensing lens L17 and a parallel light lens L18 are provided between the half mirror HM7 and the half mirror HM3.

  The laser beam reflected by the half mirror HM7 and the excitation light transmitted through the half mirror HM7 are condensed by the condensing lens L17 and converted into a parallel light beam by the parallel light lens L18, and then the half mirror of the optical axis adjusting system 500. Incident on HM3.

  Similar to the scale projection system 310 in FIG. 6, the scale projection system 320 includes a projection scale plate 5 and a mirror M2. Further, the scale projection system 320 includes a condensing lens L14, a parallel light lens L15, and a condensing lens L16 along the optical axis from the projection scale plate 5 toward the half mirror HM4 of the optical axis adjustment system 500. They are provided in this order.

  The light for scale projection from the projection scale 5 is condensed by the condensing lens L14, converted into a parallel light beam by the parallel light lens L15, and condensed by the condensing lens L16, and then the optical axis adjustment system 500. Is incident on the half mirror HM4. The scale projection system 320 may be configured not to be provided when it is not necessary to capture a scale image.

  The laser light irradiation system 400 includes an optical system such as an irradiation laser light source 41, a spatial light modulation module 42, and a cubic half mirror 44. A condensing lens L19 and a parallel light lens L20 are provided in this order along the optical axis from the irradiation laser light source 41 to the cubic half mirror 44 between the irradiation laser light source 41 and the cubic half mirror 44. ing. The laser light from the irradiation laser light source 41 is condensed by the condensing lens L19, converted into a parallel light beam by the parallel light lens L20, and then incident on the spatial light modulation module 42 via the cubic half mirror 44. The laser light reflected by the spatial light modulation module 42 is incident on the parallel light lens L21 provided between the cubic half mirror 44 and the half mirror HM7 of the illumination system 230 via the cubic half mirror 44. The parallel light lens L21 guides the incident laser light to the half mirror HM7.

  Next, the operation of the laser beam irradiation apparatus according to the present embodiment will be described. Here, an explanation will be given focusing on the operation at the time of irradiating the observation target 6 with laser light, and other operations will be omitted as appropriate.

  The spatial light modulation module 42 reads information on the fluorescent image captured by the imaging system 100 from the image recording unit 13 of the imaging apparatus 1 and stores a spatial modulation pattern based on this information.

  Thereafter, the irradiation laser light source 41 emits an irradiation laser beam in accordance with an operator's instruction. The laser light is spatially modulated in a predetermined pattern by the spatial light modulation module 42 and then enters the half mirror HM7 of the illumination system 230. The laser beam reflected by the half mirror HM7 is guided to the observation object 6 along the same optical path as the excitation light. As a result, the observation target 6 is irradiated with laser light that has been spatially modulated in a predetermined pattern based on the fluorescence image.

  As described above, in the present embodiment, the laser light from the irradiation laser light source 41 passes through the optical path along the imaging axis of the imaging system 100 in the same manner as the excitation light from the excitation light generation filter 3. Is incident on. For this reason, it is possible to accurately irradiate laser light according to the fluorescence generation pattern from the observation target 6.

  Note that a fluorescent image or the like may be captured while the observation target 6 is irradiated with laser light. At this time, in the imaging system 100, in addition to the fluorescence from the observation target 6, the laser light irradiated on the observation target 6 is also imaged simultaneously. For this reason, it can be confirmed whether the laser beam is irradiated to the desired part on the observation object 6.

  For example, when the laser beam irradiation apparatus according to the present embodiment is used for PDT treatment, in addition to confirming the laser beam irradiation site, the treatment status can also be confirmed. That is, it can be confirmed whether or not the tumor site to be treated has been destroyed by the irradiation of the laser beam.

  Thus, when imaging is performed in a state where the observation target 6 is irradiated with laser light, it is desirable to dispose the laser light attenuation filter 43 in the imaging system 100. This laser light attenuation filter 43 is provided on the upstream side or downstream side of the excitation light removal filter 4 along the imaging axis, and attenuates the laser light reflected by the observation target 6. By changing the attenuation factor of the laser light attenuating filter 43, the intensity ratio between the fluorescence incident on the image sensor 11 and the laser light can be adjusted appropriately. In particular, when the intensity of the fluorescence generated from the observation target 6 is weak with respect to the intensity of the laser beam, it is possible to prevent the fluorescence from being buried in the laser beam and being unable to be observed.

  As such a laser beam attenuation filter 43, for example, an optical filter that attenuates the laser beam intensity in a predetermined wavelength region can be used. This predetermined wavelength range is selected so as to include the wavelength range of the laser light emitted from the irradiation laser light source 41.

It is the side view which showed typically the structure of 1st Embodiment of the fluorescence observation apparatus by this invention. It is the graph which showed an example of the excitation light spectrum and the fluorescence spectrum in the case of using a fluorescence observation apparatus for PDD. It is the figure which showed the example of the image which can be observed when using a scale projection means. It is a block diagram of 2nd Embodiment of the fluorescence observation apparatus by this invention. It is a block diagram of 3rd Embodiment of the fluorescence observation apparatus by this invention. It is a block diagram of 4th Embodiment of the fluorescence observation apparatus by this invention. It is the figure which showed a part of structure of 5th Embodiment of the fluorescence observation apparatus by this invention. It is the top view which showed typically the structure of 1st Embodiment of the laser beam irradiation apparatus by this invention. It is the figure which showed the structural example of the spatial light modulation module. It is the figure which showed the structural example of the laser beam irradiation apparatus. It is the figure which showed another structural example of the laser beam irradiation apparatus. It is a block diagram of 2nd Embodiment of the laser beam irradiation apparatus by this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Imaging device, 2 ... Illuminating device, 22 ... Strobe light source, 3 ... Excitation light generation filter, 4 ... Excitation light removal filter, 5 ... Projection scale board, 10 ... Fluorescence observation apparatus, 41 ... Irradiation laser light source, 42 ... Spatial light modulation module

Claims (5)

An imaging means for capturing an image of fluorescence from an observation target;
Illuminating means having a strobe light source that emits illumination light to the observation object;
Excitation that is provided at a position on the observation target side with respect to the illuminating unit, extracts light having a predetermined excitation wavelength from the illuminating light from the illuminating unit, and generates excitation light for generating the fluorescence. Light generating means;
A light removing unit that is provided at a position on the observation target side with respect to the imaging unit and removes light of the predetermined excitation wavelength;
A fluorescence observation apparatus. The fluorescence observation apparatus according to claim 1, wherein an imaging device in which the imaging unit and the illumination unit are integrally provided in advance is used as the imaging unit and the illumination unit. The excitation light generation means has a first filter that transmits light of the predetermined excitation wavelength and is detachable from the illumination means,
The light removing means has a second filter that removes light of the predetermined excitation wavelength and is detachable from the imaging means.
The fluorescence observation apparatus according to claim 1, wherein: Scale projection means for projecting a scale onto the observation object by the illumination light from the illumination means;
The imaging means captures an image of the fluorescence and an image of the scale projected on the observation target.
The fluorescence observation apparatus according to any one of claims 1 to 3. The fluorescence observation apparatus according to any one of claims 1 to 4,
A laser light source that emits laser light for irradiating the observation object;
Spatial light modulation means that is provided on the optical path of the laser light that travels from the laser light source to the observation target, and that spatially modulates the laser light based on information on the image of the fluorescence imaged by the imaging means;
A laser beam irradiation apparatus comprising:

Images