WO2022209262A1

WO2022209262A1 - Lighting device for biological specimen observation device, biological specimen observation device, lighting device for observation device, and observation system

Info

Publication number: WO2022209262A1
Application number: PCT/JP2022/004051
Authority: WO
Inventors: 航松井; 智之大木
Original assignee: ソニーグループ株式会社
Priority date: 2021-03-31
Filing date: 2022-02-02
Publication date: 2022-10-06
Also published as: JP2022157546A

Abstract

Provided is a lighting device for a biological specimen observation device. The lighting device includes: a plurality of light-emitting elements (522); and a phosphor layer (524) which absorbs excitation light from the plurality of light-emitting elements and radiates fluorescent light. The lighting device also comprises: a light source (500) which irradiates light onto a biological specimen; and an optical filter (420) disposed between the light source and the biological specimen. The optical filter transmits the fluorescent light, and cuts at least some of the excitation light having a shorter wavelength than the fluorescent light.

Description

Illumination device for biological sample observation device, biological sample observation device, illumination device for observation device, and observation system

The present disclosure relates to an illumination device for a biological sample observation device, a biological sample observation device, an illumination device for an observation device, and an observation system.

In recent years, as a pathological diagnosis method responsible for the definitive diagnosis of diseases, in addition to the conventional method of making a pathological diagnosis while confirming a pathological specimen (biological sample) with a microscope (observation device), the pathological specimen is imaged with a scanner and an image is obtained. A method of digitizing and displaying the image and referring to the image to make a pathological diagnosis has been proposed. In addition, there is a demand for an illumination device capable of illuminating the entire observation area of a pathological specimen with light having a high color rendering property so that information necessary for diagnosis is not overlooked in order to perform an appropriate pathological diagnosis. .

　Lamp light sources (halogen lamps and xenon lamps) were widely used as light sources for lighting equipment for observing pathological specimens due to their good color rendering properties and high brightness. However, since such a lamp light source has a short life and thus requires a running cost, a white LED (Light Emitting Diode) light source has come to be used in recent years.

JP 2011-41156 A

However, when considering the use of the LED light source as a light source for observing pathological specimens, there is a significant difference in the color temperature of the LED light source due to manufacturing variations (individual differences) and aging deterioration in long-term use. It can be said that there is room for improvement.

Therefore, the present disclosure proposes an illuminating device for a biological sample observation device, a biological sample observation device, an illuminating device for an observation device, and an observation system that can suppress color temperature changes due to individual differences and aging deterioration.

According to the present disclosure, a light source for irradiating a biological sample with light, the light source and the an optical filter provided between the biological sample and the biological sample, wherein the optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence. An illumination device for a viewing device is provided.

Further, according to the present disclosure, there is provided a biological sample observation apparatus including an illumination device for illuminating a biological sample, wherein the illumination device includes a plurality of light-emitting elements, and fluorescence emitted from the plurality of light-emitting elements by absorbing excitation light emitted from the plurality of light-emitting elements. and a light source for irradiating the biological sample with light; and an optical filter provided between the light source and the biological sample, wherein the optical filter transmits the fluorescence. and cuts at least part of the excitation light having a shorter wavelength than the fluorescence.

Further, according to the present disclosure, a light source that includes a plurality of light emitting elements and a phosphor layer that absorbs excitation light from the plurality of light emitting elements and emits fluorescence, and irradiates a sample with light; an optical filter provided between the sample and the sample, wherein the optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence. A lighting device is provided.

Furthermore, according to the present disclosure, an observation device that observes a biological sample, and a computer that controls the observation device and processes signals obtained from the observation device, the observation device includes a plurality of light emitting elements and , a phosphor layer that absorbs the excitation light from the plurality of light emitting elements and emits fluorescence, a light source that irradiates the biological sample with light, and an optical device provided between the light source and the biological sample Observation comprising a filter and an imaging unit for imaging the biological sample, wherein the optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence. A system is provided.

1 is a block diagram showing a configuration example of an observation system 10 according to an embodiment of the present disclosure; FIG. 2 is a diagram showing a configuration example of an illumination unit 102 shown in FIG. 1; FIG. 3 is a diagram showing a configuration example of a light source 500 shown in FIG. 2; FIG. FIG. 5 is a diagram showing a configuration example of a light source 500a according to a comparative example; FIG. 5 is a diagram showing a spectrum distribution of emitted light from a light source 500a according to a comparative example; FIG. 5 is a diagram showing differences in spectral distribution of emitted light due to individual differences in a light source 500a according to a comparative example; FIG. 5 is a diagram showing changes in spectrum distribution of radiated light due to aged deterioration of a light source 500a according to a comparative example; 1 is a diagram showing a configuration example of a light source 500 according to an embodiment of the present disclosure; FIG. FIG. 5 shows a spectral distribution of emitted light from a light source 500 according to an embodiment of the present disclosure; 4A and 4B are diagrams showing transmittance characteristics of an optical filter 420 according to an embodiment of the present disclosure; 1 is a diagram (part 1) showing a configuration example of an optical system 400 according to an embodiment of the present disclosure; FIG. FIG. 2 is a diagram (part 2) showing a configuration example of an optical system 400 according to an embodiment of the present disclosure; FIG. 2 is a diagram (Part 1) showing the spectral distribution of illumination light from the illumination unit 102 according to the embodiment of the present disclosure; FIG. 5 is a diagram showing differences in spectral distribution of emitted light due to individual differences in the light source 500 according to the embodiment of the present disclosure; FIG. 2 is a diagram (part 2) showing the spectral distribution of illumination light from the illumination unit 102 according to the embodiment of the present disclosure; FIG. 5 is a diagram showing changes in the spectral distribution of emitted light due to aging of the light source 500 according to an embodiment of the present disclosure; FIG. 3 is a diagram (Part 3) showing the spectral distribution of illumination light from the illumination unit 102 according to the embodiment of the present disclosure; FIG. 10 is a diagram showing transmittance characteristics of an optical filter 420 according to a modified example of the embodiment of the present disclosure; It is a figure which shows roughly the whole structure of a microscope system. It is a figure which shows the example of an imaging system. It is a figure which shows the example of an imaging system. 1 is a block diagram showing an example of a schematic configuration of a diagnostic support system; FIG.

Preferred embodiments of the present disclosure will be described in detail below with reference to the accompanying drawings. In the present specification and drawings, constituent elements having substantially the same functional configuration are denoted by the same reference numerals, thereby omitting redundant description. In addition, in this specification and drawings, a plurality of components having substantially the same or similar functional configuration may be distinguished by attaching different alphabets after the same reference numerals. However, when there is no particular need to distinguish between a plurality of components having substantially the same or similar functional configurations, only the same reference numerals are used.

In the following description, a tissue section or cell that is part of a tissue (eg, organ or epithelial tissue) obtained from a living body (eg, human body, plant, etc.) is referred to as a biological sample. In addition, the biological sample described below may be subjected to various staining as necessary. In other words, in each of the embodiments described below, unless otherwise specified, the biological sample does not have to be dyed in various ways. Furthermore, for example, staining includes not only general staining represented by HE (hematoxylin-eosin) staining, Giemsa staining or Papanicolaou staining, but also periodic acid-Schiff (PAS) staining used when focusing on a specific tissue. and fluorescent staining such as FISH (Fluorescence In-Situ Hybridization) and enzyme antibody method.

Also, the description shall be given in the following order.
1. Background leading to creation of embodiments of the present disclosure 1.1 Observation system 1.2 Scanner 1.3 Illumination unit 1.4 Light source 1.5 Background 2. Embodiment 2.1 Light source 2.2 Optical filter 2.3 Illumination light 2.4 Modification 3. Summary 4. Application examples 4.1 Microscope system 4.2 Pathological diagnosis system 5. supplement

<<1. Background leading to the creation of the embodiments of the present disclosure>>
<1.1 Observation system>
First, before describing the embodiment of the present disclosure, a configuration example of an observation system 10 according to the first embodiment of the present disclosure will be described with reference to FIG. FIG. 1 is a block diagram showing a configuration example of an observation system 10 according to an embodiment of the present disclosure. The observation system 10 according to this embodiment is a scanner system that digitally photographs a slide 300 on which a biological sample (for example, cell tissue or the like) is mounted.

As shown in FIG. 1, an observation system 10 according to the present embodiment can include a scanner (observation device) 100 and an image processing device 200 . Note that the scanner 100 and the image processing apparatus 200 may be communicatively connected to each other via various wired or wireless communication networks. Further, the number of scanners 100 and image processing devices 200 included in the observation system 10 according to the present embodiment is not limited to the number illustrated in FIG. 1, and may include more. Furthermore, the observation system 10 according to this embodiment may include other servers, devices, and the like (not shown). Below, an outline of each device included in the observation system 10 according to the present embodiment will be described.

(Scanner 100)
The scanner 100 irradiates the slide 300 of the biological sample placed on the stage 108 of the scanner 100 with predetermined illumination light, and emits light transmitted through the slide 300 or light emitted from the slide 300. can be photographed (imaged). For example, the scanner 100 can be a microscope including a magnifying glass (not shown) and a digital camera (not shown) that can magnify and photograph a biological sample. Note that the scanner 100 may be implemented by any device having a photographing function, such as a smartphone, tablet, game machine, or wearable device. Further, the scanner 100 is driven and controlled by an image processing device 200, which will be described later, and the image captured by the scanner 100 is stored in the image processing device 200, for example. A detailed configuration of the scanner 100 will be described later.

(Image processing device 200)
The image processing device 200 is a device having a function of controlling the scanner 100 and processing an image (signal) captured by the scanner 100 . Specifically, the image processing apparatus 200 controls the scanner 100 to capture a digital image of the biological sample, and performs predetermined image processing on the obtained digital image. The image processing device 200 is realized by any device having a control function and an image processing function, such as a PC (Personal Computer), a tablet, a smartphone, or the like.

Note that in the present embodiment, the scanner 100 and the image processing device 200 may be integrated devices, that is, they may not be realized by a single device. Further, in this embodiment, the scanner 100 and the image processing apparatus 200 described above may be realized by a plurality of devices that are connected via various wired or wireless communication networks and cooperate with each other. Furthermore, the image processing apparatus 200 described above can be realized by, for example, a hardware configuration of a computer described later.

<1.2 Scanner>
Further, the detailed configuration of the scanner 100 according to this embodiment will be described with reference to FIG. As shown in FIG. 1, the scanner 100 can mainly have an illumination section (illumination device) 102, a sensor section (imaging section) 104, a control section 106, and a stage . Each functional block of the scanner 100 will be sequentially described below.

(Illumination unit 102)
The illumination unit 102 is provided on the side of the stage 108 opposite to the slide placement surface on which the slide 300 can be placed, and irradiates the slide 300 of the biological sample with illumination light under the control of the control unit 106, which will be described later. It is a lighting device that can Also, the illumination unit 102 may have, for example, a lens (optical system) (not shown) that collects the illumination light emitted from the illumination unit 102 and guides it to the slide 300 on the stage 108 . A detailed configuration of the illumination unit 102 will be described later.

(Sensor unit 104)
The sensor unit 104 is provided on the side of the slide arrangement surface of the stage 108, and is a color sensor that detects, for example, red (R), green (G), and blue (B) light, which are the three primary colors. More specifically, the sensor unit 104 can have, for example, an objective lens (not shown) and an imaging element (not shown). Then, the sensor unit 104 can digitally photograph (image) the biological sample and output the photographed digital image to the image processing apparatus 200 under the control of the control unit 106 to be described later.

Specifically, the objective lens (not shown) is provided on the side of the slide arrangement surface of the stage 108, and makes it possible to magnify and photograph the biological sample. That is, the transmitted light transmitted through the slide 300 arranged on the stage 108 is condensed by the objective lens, and the imaging element (illustrated omitted).

The imaging device (not shown) has a photographing range having a predetermined horizontal width and vertical width on the slide arrangement surface of the stage 108 according to the pixel size of the imaging device and the magnification of the objective lens (not shown). An image is formed. It should be noted that when part of the biological sample is magnified by the objective lens, the imaging range described above is sufficiently narrower than the imaging range of the imaging element. More specifically, the imaging element can be realized by an imaging element such as a CCD (Charge Coupled Device) or a CMOS (Complementary Metal Oxide Semiconductor).

In this embodiment, the sensor unit 104 may directly photograph the biological sample without using an objective lens or the like, or may photograph the biological sample via an objective lens or the like, and is not particularly limited. do not have.

(control unit 106)
The control unit 106 can comprehensively control the operation of the scanner 100, and includes processing circuits realized by, for example, a CPU (Central Processing Unit), a ROM (Read Only Memory), a RAM (Random Access Memory), and the like. . For example, the control unit 106 can control the lighting unit 102 and the sensor unit 104 described above. Further, the controller 106 may control a stage drive mechanism (not shown) that moves the stage 108 in various directions.

For example, the control unit 106 may control the number of shots N and the shooting time of the sensor unit 104 according to commands output from the image processing device 200 . More specifically, the control unit 106 may control the sensor unit 104 to intermittently perform imaging N times at predetermined intervals. Also, the control unit 106 may control the wavelength, irradiation intensity, or irradiation time of the illumination light emitted from the illumination unit 102 . Furthermore, the control unit 106 controls a stage drive mechanism (not shown) that moves the stage 108 in various directions according to the region of interest so that a preset region of interest (ROI) is imaged. good too. The term "region of interest" as used herein means a region (target region) of a biological sample that a user pays attention to for analysis or the like.

(Stage 108)
The stage 108 is a mounting table on which the slide 300 is mounted. Further, the stage 108 may be provided with a stage drive mechanism (not shown) for moving the stage 108 in various directions. For example, by controlling the stage drive mechanism, the stage 108 is freely moved in a direction parallel to the slide arrangement surface (X-axis-Y-axis direction) and in a direction orthogonal to the slide arrangement surface (Z-axis direction). be able to. Further, in this embodiment, the stage 108 may be provided with a sample transport device (not shown) that transports the slide 300 to the stage 108 . By providing such a transport device, the slide 300 to be photographed can be automatically placed on the stage 108, and the replacement of the slide 300 can be automated.

<1.3 Lighting unit>
Next, the basic configuration of the lighting unit 102 according to this embodiment will be described with reference to FIG. FIG. 2 is a diagram showing a configuration example of the lighting unit 102 shown in FIG. For example, as shown in FIG. 2, the illumination unit 102 includes a plurality of lenses to irradiate a slide 300 on which a biological sample such as a pathological specimen is mounted with illumination light that is uniform and has high color rendering properties. It includes an optical system 400 including 402 and the like, and a light source 500 . Details of each block of the illumination unit 102 will be described below.

The optical system 400, as shown in FIG. 2, is a Koehler illumination composed of

lenses

402a, 402b, and 402c, a field stop 412, and an aperture stop 414.

Specifically, in the Koehler illumination, the focus of the condenser lens (lens 402c in FIG. 2) passes through the condenser lens (lens 402a in FIG. 2) and the relay lens (lens 402b in FIG. 2) to the light source ( In FIG. 2, the condenser lens, condenser lens, and relay lens are arranged so as to coincide with the point where the image of the light source 500) is formed, and furthermore, the aperture stop 414 is arranged at the coincident point. . The condenser lens and the relay lens are arranged so that the focal point of the relay lens coincides with the point where the image of the object is formed through the condenser lens, and the field stop 412 is arranged between the condenser lens and the relay lens. be done. Such Koehler illumination not only irradiates uniform light but also does not directly irradiate the light from the light source, so the distance between the subject and the light source can be increased. The impact can be suppressed. Further, in Koehler illumination, the magnification can be adjusted by adjusting the lens or the like, so the light source can be made small.

More specifically, the optical system 400 will be described in order from the light source 500 side in FIG. can be done. A field stop 412 is provided above the lens 402a and can adjust the illumination range. Furthermore, since the field stop 412 cuts unnecessary light, it is possible to suppress the occurrence of flare, ghost, etc., and obtain a clear field of view. Also, the lens 402b is provided above the field stop 412 and can converge the substantially parallel light. An aperture stop 414 is provided above the lens 402b and can adjust the brightness. Furthermore, the lens 402c is provided above the aperture stop 414, and can make the condensed light into substantially parallel light again.

Note that the optical system 400 is not limited to the Kohler illumination as described above, and may be critical illumination that does not include the condenser lens 402c.

(Light source 500)
The light source 500 is a white LED (Light Emitting Diode) lighting device that emits white light. A detailed configuration of the light source 500 will be described later.

<1.4 Light source>
Next, the basic configuration of the light source 500 according to this embodiment will be described with reference to FIG. FIG. 3 is a diagram showing a configuration example of the light source 500 shown in FIG. For example, the light source 500 has a housing 510, a plurality of LED chips (light emitting elements) 522, and a phosphor layer 524, as shown in FIG. Details of each block of the light source 500 will be described below.

(Case 510)
Specifically, the housing 510 has an open top surface, and a plurality of LED chips 522 are mounted on the inner side of the bottom surface (substrate) facing the top surface.

(LED chip 522)
The LED chip 522 is a diode that emits light when a voltage is applied, and includes an electrode (not shown) provided on a semiconductor substrate (not shown), a light emitting layer (not shown), and the like. Furthermore, the light source 500 preferably has multiple LED chips 522 to increase brightness. Details of the LED chip 522 according to the present embodiment will be described later.

(Phosphor layer 524)
The phosphor layer 524 is provided above the plurality of LED chips 522 and can absorb excitation light from the LED chips 522 and emit fluorescence in a wavelength range different from that of the excitation light. The phosphor layer 524 contains one or more kinds of phosphors, and preferably contains plural kinds of phosphors in order to improve the color rendering properties. In addition, a plurality of types of phosphors emit light in different wavelength ranges. The details of the phosphor layer 524 according to this embodiment will be described later.

<1.5 Background>
Next, the background leading to the creation of the embodiments of the present disclosure by the present inventors will be described with reference to FIGS. 4 to 7. FIG. FIG. 4 is a diagram showing a configuration example of a light source 500a according to a comparative example, and FIG. 5 is a diagram showing a spectral distribution of emitted light from the light source 500a according to a comparative example. Also, FIG. 6 is a diagram showing differences in spectral distribution of radiated light due to individual differences in the light source 500a according to the comparative example, and FIG. It is a figure which shows the change of. Here, the comparative example means the lighting unit 102 and the light source 500 that were repeatedly studied by the present inventors before the embodiment of the present disclosure.

As described above, in recent years, as a pathological diagnosis method for definitive diagnosis of a disease, a method has been proposed in which a pathological specimen is imaged by the scanner 100, the image is digitized and displayed, and the image is referred to for pathological diagnosis. It is In addition, the illumination unit 102 of the scanner 100 for performing pathological diagnosis has a color-rendering property for the entire observation area of the pathological specimen so that information necessary for diagnosis is not overlooked in order to appropriately perform a pathological diagnosis. Illumination light having a high optical spectrum (for example, a Color Rendering Index (CRI) of 90 or higher) is required. The color rendering properties (color shades) of the illumination unit 102 are directly related to how pathological specimens are viewed, and are therefore a very important factor in pathological diagnosis. Color rendering refers to the properties of a lighting device, etc., that affect how the color of an object appears when the lighting device, etc. irradiates light onto the object. It is said that it is preferable to be close to the direction. More specifically, the color rendering property is indicated by an index called CRI. CRI is 100 in sunlight, and CRI approaches 100 as the appearance of an object is closer to that in sunlight.

Further, until now, lamp light sources (halogen lamps and xenon lamps) have been widely used as the light source 500 of the illumination unit 102 due to their good color rendering properties and high luminance. However, as described above, such a lamp light source has a short life and thus requires high running costs. Therefore, in recent years, white LED light sources have come to be used.

For example, as shown in FIG. 4, a light source 500a according to a comparative example that has been studied until now has excitation light (an arrow corresponds), and the fluorescence emitted from the phosphor layer 524a (specifically, including a yellow phosphor that emits yellow light) that has absorbed the excitation light (corresponds to the short arrow shown in the figure) Mixed white light can be emitted. Specifically, the radiated light emitted from the light source 500a according to the comparative example exhibits a spectral distribution as shown in FIG. More specifically, the light source 500a according to the comparative example emits excitation light with a peak wavelength of around 450 nm and fluorescent light with a wavelength range of about 500 nm to about 650 nm. The CRI of the light source 500a according to the comparative example is about 70.

Furthermore, in the light source 500a according to the comparative example, due to manufacturing variations (individual differences) of the LED chips 522a and deterioration over time of the LED chips 522a and the phosphor contained in the phosphor layer 524 during long-term use, the color temperature of the light source 500a is reduced to about 100K. I know it makes a difference. Specifically, as shown in FIG. 6, the peak wavelength (450 nm±5 nm) of the excitation light varies due to manufacturing variations (individual differences) of the LED chips 522a. It is 6510K (median value 6652K), and the maximum difference is 100K or more.

In addition, due to deterioration over time of the LED chip 522a and the phosphor, the intensity of the excitation light component (peak wavelength 450 nm) changes. It has been found that a degree of color temperature variation occurs. Specifically, as shown in FIG. 7, due to aging, the range of color temperature variation is from 5812K (aging deterioration of the LED chip 522) to 7440K (aging deterioration of the phosphor) (initial value 6652K). A difference of several thousand K or more will exist.

In this way, since there are significant differences due to individual differences and aging deterioration in the color temperature of the light source 500a, for example, when digitizing an image of a pathological specimen, color correction for each light source 500a and color correction corresponding to aging are required. A correction will be required. However, since there is a limit to making the appearance constant by correction, it has been required to suppress such color temperature changes due to individual differences and aging deterioration.

Therefore, the present inventors diligently studied the light source 500 in such a situation and independently obtained the knowledge that the difference in color temperature of the light source 500 is mainly due to the change in the excitation light component. Based on such findings, the present inventors came up with the idea of using an optical filter that appropriately cuts the excitation light component. According to the embodiments of the present disclosure created by the present inventors, by using such an optical filter, it is possible to reduce the effects of changes in the components of the excitation light. can be obtained. Hereinafter, the details of the embodiments of the present disclosure created by the present inventors will be sequentially described.

<<2. Embodiment>>
<2.1 Light source>
First, the detailed configuration of the light source 500 according to the embodiment of the present disclosure will be described with reference to FIGS. 8 and 9. FIG. FIG. 8 is a diagram showing a configuration example of the light source 500 according to this embodiment, and FIG. 9 is a diagram showing a spectral distribution of emitted light from the light source 500 according to this embodiment. The light source 500 according to this embodiment is a lighting device that emits white light, and has a CRI of 90 or higher.

Specifically, the light source 500 has a housing 510, a plurality of LED chips (light emitting elements) 522, and a phosphor layer 524, as shown in FIG. Details of each block of the light source 500 according to the present embodiment will be described below.

(Case 510)
The housing 510 has an open top surface, and an LED chip 522 is mounted on the inner side of the bottom surface (substrate) facing the top surface, as in the comparative example described above.

(LED chip 522)
As in the comparative example, the LED chip 522 is a diode that emits light when a voltage is applied, and includes an electrode (not shown) and a light emitting layer (not shown) provided on a semiconductor substrate (not shown). Furthermore, the light source 500 according to this embodiment preferably has a plurality of LED chips 522 to increase brightness.

Furthermore, in the present embodiment, unlike the comparative example (which has a peak wavelength of excitation light of 450 nm), the LED chip 522 emits excitation light having a peak wavelength of 425 nm or less, which is a wavelength band from violet to blue-violet ( corresponding to the thick arrow). For example, the LED chip 522 emits excitation light with a peak wavelength of 420 nm. In this embodiment, the LED chip 522 preferably emits excitation light that does not easily affect the color rendering properties of the light source 500, and has a wavelength band separate from the wavelength band of fluorescence emitted from the phosphor layer 524 described later. It is preferable to emit excitation light with

(Phosphor layer 524)
As in the comparative example, the phosphor layer 524 is provided above the plurality of LED chips 522, as shown in FIG. can radiate. In this embodiment, the phosphor layer 524 includes a plurality of phosphors (not shown) that emit fluorescence in different wavelength ranges. For example, each phosphor emits red light, green light, and blue light. , respectively (corresponding to the thin arrows in FIG. 8).

Specifically, the radiated light emitted from the light source 500 according to this embodiment exhibits a spectral distribution as shown in FIG. More specifically, the light source 500 emits excitation light with a peak wavelength of 420 nm and fluorescent light with a wavelength band from about 450 nm to about 650 nm, which is longer than the excitation light.

<2.2 Optical Filter>
Details of the optical filter 420 according to an embodiment of the present disclosure will now be described with reference to FIGS. 10-12. FIG. 10 is a diagram showing transmittance characteristics of the optical filter 420 according to this embodiment, and FIGS. 11 and 12 are diagrams showing configuration examples of the optical system 400 according to this embodiment.

In this embodiment, an optical filter 420 (see FIGS. 11 and 12) is provided between the light source 500 and the slide 300 on which the biological sample is mounted. The optical filter 420 can transmit fluorescence from the light source 500 and cut at least part of the excitation light from the light source 500 . In the present embodiment, the excitation light has a wavelength band that is less likely to affect the color rendering properties and is separate from the wavelength band of the fluorescence emitted from the phosphor layer 524. Therefore, the optical filter 420 is can be transmitted, and the excitation light can be cut.

Specifically, in this embodiment, the optical filter 420 preferably cuts at least part of the peak wavelength component of the excitation light. It is preferable to have a transmittance of 50% or less at a wavelength longer than the peak wavelength by a predetermined wavelength. Furthermore, in the present embodiment, it is preferable that the predetermined wavelength is set to, for example, about 10 nm in consideration of the variation.

Specifically, as shown in FIG. 10, the optical filter 420 has, for example, a transmittance of 50% or less (specifically, 38.4%) at 430 nm, and transmits light with a wavelength longer than 430 nm. be able to. For example, an optical filter having such transmittance characteristics is a UV (ultraviolet) cut filter.

Furthermore, in the present embodiment, a lens 402a made of a collimating lens is provided on the side of the light source 500 of the optical system 400 included in the illumination unit 102 so that the light beam from the light source 500 becomes substantially parallel light. The optical filter 420 can be provided between the light source 500 and the lens 402a, as shown in FIG. is narrower, an effect of suppressing chromatic aberration can be expected. Alternatively, the optical filter 420 can be provided between the lens 402a and the slide 300, as shown in FIG. In such a case, the lens 402a narrows the incident angle distribution of the light rays incident on the optical filter 420, so that it is expected to suppress the influence of the difference in the transmission characteristics of the optical filter due to the difference in the incident angle of the light. can.

Although the optical system 400 shown in FIGS. 11 and 12 has the above-described Koehler illumination configuration, the present embodiment is not limited to such Koehler illumination, and may be critical illumination. . Specifically, the optical system 400 includes an aperture stop 414 provided between the light source 500 and the slide 300 , between the light source 500 and the aperture stop 414 , and between the aperture stop 414 and the slide 300 . Critical illumination with multiple lenses 402 . Note that the critical illumination is an illumination method that allows the image of the light source 500 to be directly focused on the observation area. can do. However, since the image of the light source 500 can be directly focused on the observation area, illumination unevenness is likely to occur, and the subject is likely to be affected by the heat from the light source 500 .

<2.3 Illumination light>
Next, illumination light from the illumination unit 102 according to this embodiment will be described with reference to FIGS. 13 to 17. FIG. FIG. 13 is a diagram showing the spectral distribution of illumination light from the illumination unit 102 according to this embodiment. FIG. 14 is a diagram showing differences in spectral distribution of radiated light due to individual differences in the light source 500 according to this embodiment, and FIG. 15 shows differences in spectral distribution of radiated light due to individual differences as shown in FIG. FIG. 10 is a diagram showing a spectral distribution of illumination light from an illumination unit 102 when it exists. FIG. 16 is a diagram showing changes in spectral distribution of radiated light due to aged deterioration of the light source 500 according to the present embodiment, and FIG. 17 shows spectral distribution of radiated light due to aged deterioration as shown in FIG. FIG. 10 is a diagram showing the spectral distribution of illumination light from the illumination unit 102 when there is a change;

Illumination light emitted from the illumination unit 102 including the light source 500 and the optical filter 420 according to this embodiment exhibits a spectral distribution as shown in FIG. More specifically, the light source 500 cuts the excitation light component from the light source 500 by the optical filter 420 compared to the comparative example shown in FIG. Can be irradiated.

Here, in the light source 500 according to the present embodiment, as shown in FIG. 14, due to manufacturing variations (individual differences) of the LED chips 522, the peak wavelength of the excitation light varies within a range of 420 nm±5 nm. think about. Even in such a case, by using the optical filter 420 according to the present embodiment, it is possible to suppress the influence of the variations described above. Specifically, the illumination light emitted from the illumination unit 102 according to the present embodiment exhibits a spectrum distribution as shown in FIG. The difference in color temperature can be suppressed to about 10K. On the other hand, in the comparative example (not using the optical filter 420 according to the present embodiment) described above, as shown in FIG. The color temperature range is from 6510K to 6690K (the median value is 6652K), and the maximum color temperature difference is 100K or more.

Here, in the light source 500 according to the present embodiment, due to aged deterioration of the phosphor contained in the LED chip 522 and the phosphor layer 524, as shown in FIG. Consider the case where the proportions of the components vary. Specifically, in FIG. 16, the intensity of the excitation light component decreases due to deterioration of the LED chip 522, and the deterioration of the phosphor makes it difficult for the excitation light component to be absorbed by the phosphor, reducing the intensity of the excitation light component. shown to do. Even in such a case, by using the optical filter 420 according to the present embodiment, the influence of the aged deterioration can be suppressed. Specifically, the illumination light emitted from the illumination unit 102 according to the present embodiment exhibits a spectrum distribution as shown in FIG. deterioration) (initial value 4691K), and the difference in color temperature can be suppressed to about K. On the other hand, in the comparative example (not using the optical filter 420 according to the present embodiment) described above, as shown in FIG. ) to 7440K (degradation of phosphor over time) (initial value 6652K), and the difference in color temperature becomes several thousand K or more at maximum.

<2.4 Modifications>
Moreover, this embodiment may be modified, and a modification according to this embodiment will be described with reference to FIG. 18 . FIG. 18 is a diagram showing transmittance characteristics of an optical filter 420 according to a modification of this embodiment.

In this modified example, the optical filter 420 may have transmittance characteristics as shown in FIG. Specifically, as shown in FIG. 18, the optical filter 420 has a transmittance of 50% or less at 430 nm and can transmit light with wavelengths longer than 430 nm. Furthermore, the optical filter 420 has a transmittance of 50% or less (specifically 48.1%) at 700 nm. For example, an optical filter 420 having such transmittance characteristics may be a UV-IR (infrared) cut filter. Since the optical filter 420 according to this modification can also function as an IR filter for avoiding infrared light from entering the sensor unit 104, it has the advantage of suppressing an increase in the number of parts of the scanner 100 described above. have.

<<3. Summary>>
As described above, according to the embodiments of the present disclosure, by using an optical filter that appropriately cuts the components of the excitation light, it is possible to reduce the effects of changes in the components of the excitation light. It is possible to obtain the lighting unit 102 with little change in color temperature due to deterioration. As a result, according to the present embodiment, it is possible to reduce the need for color correction for each lighting unit 102 and color correction corresponding to aging when digitizing an image of a pathological specimen, for example. Furthermore, according to the present embodiment, it is possible to reduce the need for adjusting the imaging element (not shown) of the sensor unit 104 .

Note that the illumination unit (illumination device) 102 according to the present embodiment is not limited to being applied to the scanner 100 as described above. may be used as an illumination device for an optical microscope (not shown). Further, the scanner 100 is not limited to being used in the observation system 10 as described above, and may be used alone.

In addition, in the above-described embodiments of the present disclosure, observation targets are not limited to biological samples. In addition, the above-described embodiments of the present disclosure are not limited to application to medical or research applications, and industrial microscopes and the like that require high-precision analysis using images. is not particularly limited as long as it is used for

<<4. Application example >>
<4.1 Microscope system>
Also, the technology according to the present disclosure can be applied to, for example, a microscope system. A configuration example of an applicable microscope system will be described below with reference to FIG. FIG. 19 shows a configuration example of a microscope system.

A microscope system 5000 shown in FIG. 19 includes a microscope device 5100 , a control section 5110 and an information processing section 5120 . The microscope device 5100 has a light irradiation section 5101 , an optical section 5102 and a signal acquisition section 5103 . The microscope device 5100 may further have a sample placement section 5104 on which the biological sample S is placed. Note that the configuration of the microscope device is not limited to that shown in FIG. It may be used as the light irradiation unit 5101 . Further, the light irradiation section 5101 may be arranged such that the sample mounting section 5104 is sandwiched between the light irradiation section 5101 and the optical section 5102, and may be arranged on the side where the optical section 5102 exists, for example. The microscope apparatus 5100 may be configured to be able to perform one or more of bright field observation, phase contrast observation, differential interference contrast observation, polarization observation, fluorescence observation, and dark field observation.

The microscope system 5000 may be configured as a so-called WSI (Whole Slide Imaging) system or a digital pathology imaging system, and can be used for pathological diagnosis. Microscope system 5000 may also be configured as a fluorescence imaging system, in particular a multiplex fluorescence imaging system.

For example, the microscope system 5000 may be used to perform intraoperative pathological diagnosis or remote pathological diagnosis. In the intraoperative pathological diagnosis, while the surgery is being performed, the microscope device 5100 acquires data of the biological sample S obtained from the subject of the surgery, and transmits the data to the information processing unit 5120. and can be sent. In the remote pathological diagnosis, the microscope device 5100 can transmit the acquired data of the biological sample S to the information processing unit 5120 located in a place (another room, building, etc.) away from the microscope device 5100. . In these diagnoses, the information processing section 5120 receives and outputs the data. Further, the user of the information processing section 5120 can make a pathological diagnosis based on the output data.

(Biological sample S)
The biological sample S may be a sample containing a biological component. The biological components may be tissues, cells, liquid components of a living body (blood, urine, etc.), cultures, or living cells (cardiomyocytes, nerve cells, fertilized eggs, etc.). Also, the biological sample may be a solid substance, a specimen fixed with a fixing reagent such as paraffin, or a solid substance formed by freezing. The biological sample can be a section of the solid. A specific example of the biological sample is a section of a biopsy sample.

The above biological sample may be one that has undergone processing such as staining or labeling. The treatment may be staining for indicating the morphology of biological components or for indicating substances (surface antigens, etc.) possessed by biological components, such as HE (Hematoxylin-Eosin) staining, immunohistochemistry staining, and the like. can be mentioned. Also, the biological sample may have been subjected to the above treatment with one or more reagents, and the reagents may be fluorescent dyes, coloring reagents, fluorescent proteins, or fluorescently labeled antibodies.

In addition, the specimen may be one prepared from a tissue sample for the purpose of pathological diagnosis or clinical examination. Moreover, the specimen is not limited to the human body, and may be derived from animals, plants, or other materials. The type of specimen used (e.g., organs or cells, etc.), the type of target disease, the subject's attributes (e.g., age, sex, blood type, race, etc.), or the subject's lifestyle (For example, eating habits, exercise habits, smoking habits, etc.) have different properties. Therefore, the specimens may be managed with identification information (one-dimensional or two-dimensional code such as bar code or QR code (registered trademark)) that allows each specimen to be identified.

(Light irradiation unit 5101)
The light irradiation unit 5101 is a light source for illuminating the biological sample S and an optical system for guiding the light irradiated from the light source to the specimen. The light source may irradiate the biological sample with visible light, ultraviolet light, or infrared light, or a combination thereof. The light source may be one or more of a halogen light source, a laser light source, an LED light source, a mercury light source, and a xenon light source. A plurality of light source types and/or wavelengths may be used in fluorescence observation, and may be appropriately selected by those skilled in the art. The light irradiation unit 5101 can have a transmissive, reflective, or episcopic (coaxial or lateral) configuration.

(Optical section 5102)
The optical section 5102 is configured to guide the light from the biological sample S to the signal acquisition section 5103 . The optical unit 5102 can be configured to allow the microscope device 5100 to observe or image the biological sample S. Optics 5102 may include an objective lens. The type of objective lens may be appropriately selected by a person skilled in the art according to the observation method. Also, the optical unit 5102 may include a relay lens for relaying the image magnified by the objective lens to the signal acquisition unit. The optical section 5102 may further include optical components other than the objective lens and relay lens, eyepiece lens, phase plate, condenser lens, and the like. In addition, the optical section 5102 may further include a wavelength separation section configured to separate light having a predetermined wavelength from the light from the biological sample S. The wavelength separation section may be configured to selectively allow light of a predetermined wavelength or range of wavelengths to reach the signal acquisition section. The wavelength separator may include, for example, one or more of a filter that selectively transmits light, a polarizing plate, a prism (Wollaston prism), and a diffraction grating. The optical components included in the wavelength separation section may be arranged, for example, on the optical path from the objective lens to the signal acquisition section. The wavelength separation unit is provided in the microscope device when fluorescence observation is performed, particularly when an excitation light irradiation unit is included. The wavelength separator may be configured to separate fluorescent light from each other or white light and fluorescent light.

(Signal acquisition unit 5103)
The signal acquisition unit 5103 can be configured to receive light from the biological sample S and convert the light into an electrical signal, particularly a digital electrical signal. The signal acquisition unit 5103 may be configured to acquire data regarding the biological sample S based on the electrical signal. The signal acquisition unit 5103 may be configured to acquire data of an image (image, particularly a still image, a time-lapse image, or a moving image) of the biological sample S. can be configured to acquire data for an image obtained by The signal acquisition unit 5103 includes one or more imaging elements having a plurality of pixels arranged one-dimensionally or two-dimensionally, a CMOS (Complementary Metal Oxide Semiconductor), a CCD (Charge CoupLED Device), or the like. The signal acquisition unit 5103 may include an image sensor for acquiring a low-resolution image and an image sensor for acquiring a high-resolution image, or an image sensor for sensing for AF (Auto Focus) and the like and an image sensor for observation. An imaging device for image output may also be included. In addition to a plurality of pixels, the image sensor includes a signal processing unit (CPU (Central Processing Unit) that performs signal processing using pixel signals from each pixel, a DSP (Digital Signal Processor), and one or two memories. above), and an output control unit that controls output of image data generated from the pixel signals and processed data generated by the signal processing unit. Also, the imaging device including the plurality of pixels, the signal processing section, and the output control section can preferably be configured as a one-chip semiconductor device.

Note that the microscope system 5000 may further include an event detection sensor. The event detection sensor includes a pixel that photoelectrically converts incident light, and can be configured to detect, as an event, a change in luminance of the pixel exceeding a predetermined threshold. The event detection sensor may be asynchronous.

(control unit 5110)
The control unit 5110 controls imaging by the microscope device 5100 . For imaging control, the control unit 5110 can drive the movement of the optical unit 5102 and/or the sample placement unit 5104 to adjust the positional relationship between the optical unit 5102 and the sample placement unit 5104. . The control unit 5110 can move the optical unit 5102 and/or the sample mounting unit 5104 in a direction toward or away from each other (for example, the optical axis direction of the objective lens). Also, the control section 5110 may move the optical section 5102 and/or the sample placement section 5104 in any direction on a plane perpendicular to the optical axis direction. The control unit 5110 may control the light irradiation unit 5101 and/or the signal acquisition unit 5103 for imaging control.

(Sample mounting portion 5104)
The sample mounting section 5104 may be configured such that the position of the biological sample S on the sample mounting section 5104 can be fixed, and may be a so-called stage. The sample placement section 5104 can be configured to move the position of the biological sample S in the optical axis direction of the objective lens and/or in a direction perpendicular to the optical axis direction.

(Information processing unit 5120)
The information processing section 5120 can acquire data (such as imaging data) acquired by the microscope device 5100 from the microscope device 5100 . The information processing section 5120 can perform image processing on captured data. The image processing may include an unmixing process, in particular a spectral unmixing process. The unmixing process is a process of extracting data of light components of a predetermined wavelength or wavelength range from the imaging data to generate image data, or removing data of light components of a predetermined wavelength or wavelength range from the imaging data. It can include processing and the like. Further, the image processing may include an autofluorescence separation process for separating the autofluorescence component and the dye component of the tissue section, and a fluorescence separation process for separating the wavelengths between dyes having different fluorescence wavelengths. In the autofluorescence separation processing, a process of removing an autofluorescence component from the image information of the other specimen using an autofluorescence signal extracted from one of a plurality of specimens having the same or similar properties may be performed. The information processing section 5120 may transmit data for imaging control by the control section 5110, and the control section 5110 receiving the data may control imaging by the microscope apparatus 5100 according to the data.

The information processing section 5120 may be configured as an information processing device such as a general-purpose computer, and may have a CPU, RAM (Random Access Memory), and ROM (Read Only Memory). The information processing section 5120 may be included in the housing of the microscope device 5100 or may be outside the housing. Various processing or functions by the information processing section 5120 may be realized by a server computer or cloud connected via a network.

A method of imaging the biological sample S by the microscope device 5100 may be appropriately selected by a person skilled in the art according to the type of the biological sample S, the purpose of imaging, and the like. An example of the imaging method will be described below with reference to FIGS. 20 and 21. FIG. 20 and 21 are diagrams showing examples of imaging methods.

An example of the imaging method is as follows. The microscope device 5100 can first identify an imaging target region. The imaging target region may be specified so as to cover the entire region in which the biological sample S exists, or a target portion of the biological sample S (a target tissue section, a target cell, or a target lesion where the target lesion exists). may be specified to cover the Next, the microscope device 5100 divides the imaging target region into a plurality of divided regions of a predetermined size, and the microscope device 5100 sequentially images each divided region. As a result, an image of each divided area is acquired.

As shown in FIG. 20, the microscope device 5100 identifies an imaging target region R that covers the entire biological sample S. Then, the microscope device 5100 divides the imaging target region R into 16 divided regions. Then, the microscope device 5100 can image the divided region R1, and then any region included in the imaging target region R, such as a region adjacent to the divided region R1. Furthermore, the microscope device 5100 captures images of the divided areas until there are no unimaged divided areas. Note that the microscope device 5100 may also capture an area other than the imaging target area R based on the captured image information of the divided area. At this time, the positional relationship between the microscope device 5100 and the sample mounting section 5104 is adjusted in order to image the next divided area after imaging a certain divided area. The adjustment can be performed by moving the microscope device 5100, moving the sample placement section 5104, or moving both of them.

In this example, the imaging device that captures each divided area may be a two-dimensional imaging device (area sensor) or a one-dimensional imaging device (line sensor). The signal acquisition unit 5103 may image each divided area via the optical unit 5102 . In addition, the imaging of each divided region may be performed continuously while moving the microscope device 5100 and/or the sample placement unit 5104, or when imaging each divided region, the microscope device 5100 and/or Alternatively, the movement of the sample placement section 5104 may be stopped. Furthermore, the imaging target region may be divided so that the divided regions partially overlap each other, or the imaging target region may be divided so that the divided regions do not overlap. Each divided area may be imaged multiple times by changing imaging conditions such as focal length and/or exposure time.

Also, the information processing section 5120 can stitch a plurality of adjacent divided regions to generate image data of a wider region. By performing the stitching process over the entire imaging target area, it is possible to obtain an image of a wider area of the imaging target area. Further, image data with lower resolution can be generated from the image of the divided area or the image subjected to the stitching process.

Other examples of imaging methods are as follows. The microscope device 5100 can first identify an imaging target region. The imaging target region may be specified so as to cover the entire region where the biological sample S is present, or a target portion (a target tissue section or a portion where target cells are present) of the biological sample S. may be specified to cover. Next, the microscope device 5100 scans a partial region (also referred to as a “divided scan region”) of the imaging target region in one direction (also referred to as a “scanning direction”) within a plane perpendicular to the optical axis. Take an image. After completing the scan of the divided scan area, the microscope device 5100 scans the divided scan area next to the scan area. Then, the microscope device 5100 repeats these scanning operations until the entire imaging target region is imaged.

As shown in FIG. 21, the microscope device 5100 identifies the region (gray portion) where the tissue section exists in the biological sample S as the imaging target region Sa. Then, the microscope device 5100 scans the divided scan area Rs in the imaging target area Sa in the Y-axis direction. After completing scanning of the divided scan region Rs, the microscope device 5100 next scans an adjacent divided scan region in the X-axis direction. The microscope device 5100 repeats this operation until scanning is completed for the entire imaging target area Sa.

The positional relationship between the microscope device 5100 and the sample placement section 5104 is adjusted for scanning each divided scan area and for imaging the next divided scan area after imaging a certain divided scan area. The adjustment may be performed by moving the microscope device 5100, moving the sample placement section 5104, or moving both of them. In this example, the imaging device that captures each divided scan area may be a one-dimensional imaging device (line sensor) or a two-dimensional imaging device (area sensor). The signal acquisition unit 5103 may capture an image of each divided area via an enlarging optical system. Also, the imaging of each divided scan region may be performed continuously while moving the microscope device 5100 and/or the sample placement unit 5104 . The imaging target area may be divided such that the divided scan areas partially overlap each other, or the imaging target area may be divided so that the divided scan areas do not overlap. Each divided scan area may be imaged multiple times while changing imaging conditions such as focal length and/or exposure time.

Also, the information processing section 5120 can stitch a plurality of adjacent divided scan regions to generate image data of a wider region. By performing the stitching process over the entire imaging target area, it is possible to obtain an image of a wider area of the imaging target area. Further, image data with lower resolution can be generated from images of divided scan regions or images subjected to stitching processing.

<4.2 Pathological diagnosis system>
In addition, the technology according to the present disclosure is applied, for example, to a pathological diagnosis system and its support system (hereinafter referred to as a diagnosis support system) in which a doctor or the like observes cells and tissues collected from a patient and diagnoses a lesion. may This diagnosis support system may be a WSI (Whole Slide Imaging) system that diagnoses or supports diagnosis of lesions based on images acquired using digital pathology technology.

FIG. 22 is a diagram showing an example of a schematic configuration of a diagnostic support system 5500 to which the technology according to the present disclosure is applied. As shown in FIG. 22, the diagnostic support system 5500 includes one or more pathology systems 5510. A medical information system 5530 and a derivation device 5540 may also be included.

Each of the one or more pathology systems 5510 is a system mainly used by pathologists, and is installed in laboratories and hospitals, for example. Each pathology system 5510 may be installed in a different hospital, and each uses various networks such as WAN (Wide Area Network) (including the Internet), LAN (Local Area Network), public line network, and mobile communication network. It is connected to the medical information system 5530 and the derivation device 5540 via.

Each pathology system 5510 includes a microscope (specifically, a microscope used in combination with digital imaging technology) 5511, a server 5512, a display control device 5513, and a display device 5514.

The microscope 5511 has the function of an optical microscope, photographs an observation object contained in a glass slide, and acquires a pathological image, which is a digital image. Observation objects are, for example, tissues and cells collected from a patient, and may be pieces of flesh of organs, saliva, blood, and the like. For example, microscope 5511 functions as scanner 30 shown in FIG.

The server 5512 stores and saves pathological images acquired by the microscope 5511 in a storage unit (not shown). Further, when receiving a viewing request from the display control device 5513 , the server 5512 searches for pathological images from a storage unit (not shown) and sends the searched pathological images to the display control device 5513 .

The display control device 5513 sends to the server 5512 the pathological image viewing request received from the user. Then, the display control device 5513 displays the pathological image received from the server 5512 on the display device 5514 using liquid crystal, EL (Electro-Luminescence), CRT (Cathode Ray Tube), or the like. Note that the display device 5514 may be compatible with 4K or 8K, and is not limited to one device, and may be a plurality of devices.

Here, if the observation target is a solid object such as a piece of flesh of an organ, the observation target may be, for example, a stained slice. A sliced piece may be produced, for example, by slicing a block piece excised from a specimen such as an organ. Also, when slicing, the block pieces may be fixed with paraffin or the like.

Various types of staining are available for staining thin sections, including general staining that indicates the morphology of tissues such as HE (Hematoxylin-Eosin) staining, immunostaining that indicates the immune status of tissues such as IHC (Immunohistochemistry) staining, and fluorescent immunostaining. may be applied. At that time, one thin section may be stained with a plurality of different reagents, or two or more thin sections (also referred to as adjacent thin sections) continuously cut out from the same block piece may be stained with different reagents. may be dyed using

The microscope 5511 can include a low-resolution imaging unit for low-resolution imaging and a high-resolution imaging unit for high-resolution imaging. The low-resolution imaging section and the high-resolution imaging section may be different optical systems, or may be the same optical system. In the case of the same optical system, the resolution of the microscope 5511 may be changed according to the imaging target.

The glass slide containing the observation target is placed on the stage located within the angle of view of the microscope 5511. The microscope 5511 first acquires the entire image within the angle of view using the low-resolution imaging unit, and specifies the region of the observation object from the acquired entire image. Subsequently, the microscope 5511 divides the region where the observation target exists into a plurality of divided regions of a predetermined size, and sequentially captures each divided region by the high-resolution imaging unit, thereby obtaining a high-resolution image of each divided region. do. In switching the target divided area, the stage may be moved, the imaging optical system may be moved, or both of them may be moved. In addition, each divided area may overlap adjacent divided areas in order to prevent occurrence of an imaging omission area due to unintended slippage of the glass slide. Furthermore, the whole image may contain identification information for associating the whole image with the patient. This identification information may be, for example, a character string, a QR code (registered trademark), or the like.

A high-resolution image acquired by the microscope 5511 is input to the server 5512. The server 5512 divides each high resolution image into smaller size partial images (hereinafter referred to as tile images). For example, the server 5512 divides one high-resolution image into a total of 100 tile images of 10×10. At that time, if adjacent divided areas overlap, the server 5512 may perform stitching processing on adjacent high-resolution images using a technique such as template matching. In that case, the server 5512 may generate tile images by dividing the entire high-resolution image stitched together by the stitching process. However, the generation of tile images from high-resolution images may be performed before the stitching process.

Also, the server 5512 can generate tile images of smaller sizes by further dividing the tile images. Such generation of tile images may be repeated until a tile image having the size set as the minimum unit is generated.

After generating tile images of the minimum unit in this way, the server 5512 performs tile composition processing for generating a single tile image by compositing a predetermined number of adjacent tile images for all tile images. This tile synthesis process can be repeated until finally one tile image is generated. Through such processing, a pyramid-structured tile image group in which each layer is composed of one or more tile images is generated. In this pyramid structure, a tile image in one layer and a tile image in a different layer have the same number of pixels, but different resolutions. For example, when synthesizing a total of four 2×2 tile images to generate one upper layer tile image, the resolution of the upper layer tile image is half the resolution of the lower layer tile image used for synthesis. It has become.

By constructing such a pyramid-structured tile image group, it is possible to switch the level of detail of the observation target displayed on the display device, depending on the hierarchy to which the tile image to be displayed belongs. For example, when the tile image of the lowest layer is used, a narrow area of the observation object is displayed in detail, and the wider the area of the observation object is displayed more coarsely as the tile image of the upper layer is used. can.

The generated pyramid-structured tile image group is stored in a storage unit (not shown) together with, for example, identification information (referred to as tile identification information) that can uniquely identify each tile image. When the server 5512 receives a tile image acquisition request including tile identification information from another device (for example, the display control device 5513 or the derivation device 5540), the server 5512 transmits the tile image corresponding to the tile identification information to the other device. do.

Note that tile images, which are pathological images, may be generated for each imaging condition such as focal length and staining conditions. When tile images are generated for each imaging condition, a specific pathological image and other pathological images corresponding to imaging conditions different from the specific imaging condition and having the same region as the specific pathological image are generated. They may be displayed side by side. Specific shooting conditions may be specified by the viewer. Further, when a plurality of imaging conditions are designated by the viewer, pathological images of the same region corresponding to each imaging condition may be displayed side by side.

In addition, the server 5512 may store the pyramid-structured tile image group in a storage device other than the server 5512, such as a cloud server. Furthermore, part or all of the tile image generation processing as described above may be executed by a cloud server or the like.

The display control device 5513 extracts a desired tile image from the pyramid-structured tile image group according to the user's input operation, and outputs it to the display device 5514 . Through such processing, the user can obtain the feeling of observing the observation object while changing the observation magnification. That is, the display control device 5513 functions as a virtual microscope. The virtual observation magnification here actually corresponds to the resolution.

Any method may be used to capture high-resolution images. A high-resolution image may be obtained by photographing the divided areas while the stage is repeatedly stopped and moved, or a high-resolution image on the strip may be obtained by photographing the divided areas while moving the stage at a predetermined speed. good too. In addition, the process of generating tile images from high-resolution images is not an essential configuration, and by changing the resolution of the entire high-resolution image stitched together by the stitching process, images with gradual changes in resolution can be created. may be generated. Even in this case, it is possible to present the user with a step-by-step process from a low-resolution image of a wide area to a high-resolution image of a narrow area.

The medical information system 5530 is a so-called electronic medical record system, and stores information related to diagnosis, such as patient identification information, patient disease information, test information and image information used for diagnosis, diagnosis results, and prescription drugs. For example, a pathological image obtained by photographing an observation target of a certain patient can be temporarily stored via the server 5512 and then displayed on the display device 5514 by the display control device 5513 . A pathologist using the pathological system 5510 makes a pathological diagnosis based on the pathological image displayed on the display device 5514 . Pathological diagnosis results made by the pathologist are stored in the medical information system 5530 .

A derivation device 5540 may perform analysis on pathological images. A learning model created by machine learning can be used for this analysis. The derivation device 5540 may derive a classification result of a specific region, a tissue identification result, or the like as the analysis result. Further, the deriving device 5540 may derive identification results such as cell information, number, position, brightness information, and scoring information for them. These pieces of information derived by the derivation device 5540 may be displayed on the display device 5514 of the pathology system 5510 as diagnosis support information.

Note that the derivation device 5540 may be a server system configured with one or more servers (including cloud servers). Also, the derivation device 5540 may be configured to be incorporated in, for example, the display control device 5513 or the server 5512 in the pathological system 5510 . That is, various analyzes on pathological images may be performed within the pathological system 5510 .

The technology according to the present disclosure can be preferably applied to the microscope 5511 as described above among the configurations described above. By applying the technology according to the present disclosure to the microscope 5511, it is possible to obtain a clearer pathological image, and thus it is possible to more accurately diagnose a lesion.

It should be noted that the configuration described above can be applied not only to the diagnostic support system, but also to general biological microscopes such as confocal microscopes, fluorescence microscopes, and video microscopes that use digital imaging technology. Here, the object to be observed may be a biological sample such as cultured cells, fertilized eggs, or sperm, a biological material such as a cell sheet or a three-dimensional cell tissue, or a living body such as a zebrafish or mouse. Further, the object to be observed is not limited to the glass slide, and can be observed while being stored in a well plate, petri dish, or the like.

Furthermore, a moving image may be generated from still images of the observed object acquired using a microscope that utilizes digital imaging technology. For example, a moving image may be generated from still images captured continuously over a predetermined period of time, or an image sequence may be generated from still images captured at predetermined intervals. In this way, by generating moving images from still images, it is possible to observe the movements of cancer cells, nerve cells, myocardial tissue, sperm, etc. such as pulsation, elongation, and migration, and the division process of cultured cells and fertilized eggs. It becomes possible to analyze the dynamic features of objects using machine learning.

<<5. Supplement >>
Although the preferred embodiments of the present disclosure have been described in detail above with reference to the accompanying drawings, the technical scope of the present disclosure is not limited to such examples. It is obvious that a person having ordinary knowledge in the technical field of the present disclosure can conceive of various modifications or modifications within the scope of the technical idea described in the claims. are naturally within the technical scope of the present disclosure.

It should be noted that, in the embodiments of the present disclosure described above, each component of each illustrated device is functionally conceptual, and does not necessarily need to be physically configured as illustrated. In other words, the specific form of distribution and integration of each device is not limited to the one shown in the figure, and all or part of them can be functionally or physically distributed and integrated in arbitrary units according to various loads and usage conditions. Can be integrated and configured.

Also, the effects described in this specification are merely descriptive or exemplary, and are not limiting. In other words, the technology according to the present disclosure can produce other effects that are obvious to those skilled in the art from the description of this specification, in addition to or instead of the above effects.

Note that the present technology can also take the following configuration.
(1)
a light source that includes a plurality of light emitting elements and a phosphor layer that absorbs excitation light from the plurality of light emitting elements and emits fluorescence, and irradiates a biological sample with light;
an optical filter provided between the light source and the biological sample;
with
The optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence.
Illumination device for biological sample observation device.
(2)
The excitation light has a peak wavelength of 425 nm or less,
the optical filter cuts at least part of the light having the peak wavelength;
The illumination device for a biological sample observation device according to (1) above.
(3)
The illumination device for a biological sample observation device according to (2) above, wherein the optical filter has a transmittance of 50% or less at a wavelength longer than the peak wavelength by a predetermined wavelength.
(4)
The illumination device for a biological sample observation device according to (3) above, wherein the predetermined wavelength is 10 nm.
(5)
The illumination device for a biological sample observation device according to (3) or (4) above, wherein the optical filter has a transmittance of 50% or less at a wavelength of 700 nm.
(6)
The illumination device for a biological sample observation device according to any one of (1) to (5) above, wherein the phosphor layer contains a plurality of types of phosphors.
(7)
The illumination device for a biological sample observation device according to (6) above, wherein the plurality of types of phosphors respectively emit fluorescence of red light, green light, and blue light.
(8)
The illumination device for a biological sample observation device according to any one of (1) to (7) above, which is an illumination device that emits white light.
(9)
The illumination device for a biological sample observation device according to (8) above, which has a CRI of 90 or more.
(10)
The illumination device for a biological sample observation device according to any one of (1) to (9) above, wherein the plurality of light emitting elements are light emitting diodes.
(11)
The illumination device for a biological sample observation device according to any one of (1) to (10) above, further comprising a collimating lens provided between the light source and the biological sample.
(12)
The illumination device for a biological sample observation device according to (11) above, wherein the optical filter is provided between the light source and the collimating lens.
(13)
The illumination device for a biological sample observation apparatus according to (11) above, wherein the optical filter is provided between the collimator lens and the biological sample.
(14)
The illumination device for a biological sample observation device according to any one of (1) to (13) above, further comprising an optical system for guiding light from the light source to the biological sample.
(15)
The optical system is
a field stop provided between the light source and the biological sample;
an aperture stop provided between the field stop and the biological sample;
a plurality of lenses provided between the field stop and the aperture stop and between the aperture stop and the biological sample;
The illumination device for a biological sample observation device according to (14) above, comprising:
(16)
The optical system is
an aperture stop provided between the light source and the biological sample;
a plurality of lenses provided between the light source and the aperture stop and between the aperture stop and the biological sample;
The illumination device for a biological sample observation device according to (14) above, comprising:
(17)
A biological sample observation device comprising an illumination device for illuminating a biological sample,
The lighting device
a light source that includes a plurality of light emitting elements and a phosphor layer that absorbs excitation light from the plurality of light emitting elements and emits fluorescence, and irradiates the biological sample with light;
an optical filter provided between the light source and the biological sample;
has
The optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence.
Biological sample observation device.
(18)
The biological sample observation device according to (17) above, which is a microscope device.
(19)
The biological sample observation apparatus according to (17) above, further comprising an imaging unit that captures an image of the biological sample, including an imaging device.
(20)
a light source that irradiates a sample with light, including a plurality of light emitting elements and a phosphor layer that absorbs excitation light from the plurality of light emitting elements and emits fluorescence;
an optical filter provided between the light source and the sample;
with
The optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence.
Illumination device for observation equipment.
(21)
An observation device for observing a biological sample, and a computer for controlling the observation device and processing signals obtained from the observation device,
The observation device is
a light source that includes a plurality of light emitting elements and a phosphor layer that absorbs excitation light from the plurality of light emitting elements and emits fluorescence, and irradiates the biological sample with light;
an optical filter provided between the light source and the biological sample;
an imaging unit that images the biological sample;
has
The optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence.
observation system.

REFERENCE SIGNS LIST 10 observation system 100 scanner 102 illumination section 104 sensor section 106 control section 108 stage 200 image processing device 300 slide 400

optical system

402, 402a, 402b, 402c lens 412 field stop 414 aperture stop 420

optical filter

500, 500a light source 510

housing

522 ,

522a LED chip

524, 524a phosphor layer

Claims

a light source that includes a plurality of light emitting elements and a phosphor layer that absorbs excitation light from the plurality of light emitting elements and emits fluorescence, and irradiates a biological sample with light;
an optical filter provided between the light source and the biological sample;
with
The optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence.
Illumination device for biological sample observation device.
The excitation light has a peak wavelength of 425 nm or less,
the optical filter cuts at least part of the light having the peak wavelength;
The illumination device for a biological sample observation device according to claim 1.
3. The illumination device for a biological sample observation apparatus according to claim 2, wherein said optical filter has a transmittance of 50% or less at a wavelength longer than said peak wavelength by a predetermined wavelength.
The illumination device for a biological sample observation device according to claim 3, wherein said predetermined wavelength is 10 nm.
The illumination device for a biological sample observation device according to claim 3, wherein the optical filter has a transmittance of 50% or less at a wavelength of 700 nm.
The illumination device for a biological sample observation device according to claim 1, wherein the phosphor layer contains a plurality of types of phosphors.
The illumination device for a biological sample observation device according to claim 6, wherein said plurality of types of phosphors respectively emit fluorescence of red light, green light, and blue light.
The illumination device for a biological sample observation device according to claim 1, which is an illumination device that emits white light.
The illumination device for a biological sample observation device according to claim 8, wherein the CRI is 90 or higher.
The illumination device for a biological sample observation device according to claim 1, wherein the plurality of light emitting elements are composed of light emitting diodes.
The illumination device for a biological sample observation device according to claim 1, further comprising a collimating lens provided between said light source and said biological sample.
The illumination device for a biological sample observation device according to claim 11, wherein said optical filter is provided between said light source and said collimating lens.
The illumination device for a biological sample observation apparatus according to claim 11, wherein said optical filter is provided between said collimator lens and said biological sample.
The illumination device for a biological sample observation device according to claim 1, further comprising an optical system for guiding light from said light source to said biological sample.
The optical system is
a field stop provided between the light source and the biological sample;
an aperture stop provided between the field stop and the biological sample;
a plurality of lenses provided between the field stop and the aperture stop and between the aperture stop and the biological sample;
15. The illumination device for a biological sample observation device according to claim 14, comprising:
The optical system is
an aperture stop provided between the light source and the biological sample;
a plurality of lenses provided between the light source and the aperture stop and between the aperture stop and the biological sample;
15. The illumination device for a biological sample observation device according to claim 14, comprising:
A biological sample observation device comprising an illumination device for illuminating a biological sample,
The lighting device
a light source that includes a plurality of light emitting elements and a phosphor layer that absorbs excitation light from the plurality of light emitting elements and emits fluorescence, and irradiates the biological sample with light;
an optical filter provided between the light source and the biological sample;
has
The optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence.
Biological sample observation device.
The biological sample observation device according to claim 17, which is a microscope device.
The biological sample observation device according to claim 17, further comprising an imaging unit that captures an image of the biological sample, including an imaging device.
a light source that irradiates a sample with light, including a plurality of light emitting elements and a phosphor layer that absorbs excitation light from the plurality of light emitting elements and emits fluorescence;
an optical filter provided between the light source and the sample;
with
The optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence.
Illumination device for observation equipment.
an observation device for observing a biological sample, and a computer for controlling the observation device and processing signals obtained from the observation device,
The observation device is
a light source that includes a plurality of light emitting elements and a phosphor layer that absorbs excitation light from the plurality of light emitting elements and emits fluorescence, and irradiates the biological sample with light;
an optical filter provided between the light source and the biological sample;
an imaging unit that images the biological sample;
has
The optical filter transmits the fluorescence and cuts at least part of the excitation light having a shorter wavelength than the fluorescence.
observation system.