JP6546976B2 - Light observation apparatus and light observation method - Google Patents

Description

  The present invention relates to a light observation apparatus for observing a light image of an object, and an imaging apparatus used therefor.

  In technical fields such as life sciences, observation light (for example, fluorescence etc.) from a sample is dispersed according to the wavelength, and a plurality of light images dispersed as a result are observed. For example, Non-Patent Document 1 below discloses a dual view microscopy technique using one camera, in which the observation light is dispersed and dispersed by a light splitting assembly attached to the outside of the microscope body. One observation light can be taken using a camera. Further, Patent Document 1 below and Patent Document 2 below disclose light splitting optical systems used in the inspection technique of dual view.

U.S. Pat. No. 5,926,283 U.S. Patent No. 7,667,761

K. Kinosita, Jr. et al., “DualView Microscopy with a Single Camera: Real-Time Imaging of Molecular Orientations and Calcium”, The Journal of Cell Biology, Volume 115, Number 1, October 1991, pp 67-73.

  However, in the above-described conventional microscope inspection technique, two types of light images are observed by one camera, and therefore two types of light are used when a camera adopting a rolling readout method such as a CMOS camera is used. There are cases where imaging timings at specific parts of an image differ from one another. As a result, comparative observation of two types of light images for a sample tends to be difficult.

  Therefore, the present invention has been made in view of such problems, and a light observation device and an imaging device that facilitate comparative observation of two types of light images of an object even when the rolling readout method is adopted. Intended to provide.

  In order to solve the above-mentioned subject, an imaging device concerning one embodiment of the present invention divides observation light of a subject into the 1st and 2nd light from the exterior, and it is 1st and 2nd light in a predetermined mode. An imaging apparatus for use in a light observation apparatus for simultaneously imaging first and second light images generated on the basis, the first pixel array group including a plurality of pixel arrays including a plurality of pixels arranged side by side And a second pixel column group adjacent to the first pixel column group and including a plurality of pixel columns including a plurality of pixels arranged in a line, and a first signal for processing signal readout from the first pixel column group A readout circuit, a second signal readout circuit that processes signal readout from the second pixel column group independently of the first signal readout circuit, and rolling by the first signal readout circuit and the second signal readout circuit And a control unit that controls the reading operation. When the mode is selected, the signal readout in the first pixel array group by the first signal readout circuit is processed by rolling readout, and the direction of the rolling readout in the first pixel array group is a plurality of pixel arrays. The signal readout in the second pixel array group by the second signal readout circuit is processed by rolling readout, and the direction of the rolling readout in the second pixel array group is plural. It sets so that it may become one direction in the parallel direction of the pixel row of this, and is controlled so that the timing of signal readout synchronizes between the 1st pixel row group and the 2nd pixel row group.

  According to this imaging device, the first light image formed on the basis of the first light divided from the observation light of the object is divided by the first pixel row group included in the first region of the imaging device. It is possible to receive light. At the same time, a second light image formed on the basis of the second light divided from the observation light of the object is made receivable by the second group of pixel columns included in the second region of the imaging device. Ru. Then, the detection signals of the first and second light images are read by rolling independently from the first and second pixel column groups by the first and second signal readout circuits, respectively, and the first pixel column group is read. The direction of the rolling readout in the second pixel row and the direction of the rolling readout in the second pixel row are made the same. This makes it easy to align the exposure timing on the same part of the object between the detection image of the first light image and the detection image of the second light image, so the exposure conditions of each part of the two light images are uniform. Can be As a result, comparative observation using detected images of two light images can be facilitated.

  Here, the control unit is configured such that the direction of the rolling readout in the first pixel column group and the direction of the rolling readout in the second pixel column group are the same in the parallel direction of the plurality of pixel columns. The second readout mode in which the readout mode, the rolling readout direction in the first pixel column group, and the rolling readout direction in the second pixel column group are opposite in the parallel direction of the plurality of pixel columns It is also preferable to switch between With this configuration, when two light images are simultaneously captured by the imaging device, the exposure condition of each portion of the two light images can be made uniform by switching to the first readout mode. On the other hand, when a single light image is captured by the imaging device, switching to the second readout mode can prevent the occurrence of linear contrast in the detection image of the single light image. As a result, even when the observation mode for capturing two light images and the observation mode for capturing a single light image are shared, it is possible to observe the light image with high accuracy.

  According to the present invention, comparison observation of two types of light images of an object is facilitated even when the rolling readout method is adopted.

It is a schematic block diagram of light observation device 1 of one embodiment of the present invention. It is a perspective view which shows an example of a structure of the light division optical apparatus 7 of FIG. It is a block diagram which shows the structure of the imaging device 8 of FIG. It is a figure which shows notionally the direction of the rolling read-out in pixel row group 47a, 47b on the light-receiving surface 41 controlled by the control part 9 of FIG. It is a figure which shows notionally the direction of the rolling read-out in pixel row group 47a, 47b on the light-receiving surface 41 controlled by the control part 9 of FIG. It is a flowchart which shows the procedure of the light observation method which concerns on this embodiment of this invention. (A) is a figure which shows notionally the direction of the rolling readout on the light-receiving surface 41 controlled by the control part 9 at the time of use in double view mode, (b) is on the light-receiving surface 41 corresponding to it. FIG. 6 is a view showing an exposure timing in each pixel column 46. (A) is a figure which shows notionally the direction of the rolling readout on the light-receiving surface 41 controlled by the control part 9 at the time of use in single view mode, (b) is on the light-receiving surface 41 corresponding to it. FIG. 6 is a view showing an exposure timing in each pixel column 46. It is a figure which shows notionally the direction of the rolling read-out in pixel row group 47a, 47b on the light-receiving surface 41 controlled by the control part 9 which concerns on the modification of this invention. It is a figure which shows notionally the direction of the rolling read-out in pixel row group 47a, 47b on the light-receiving surface 41 controlled by the control part 9 which concerns on the modification of this invention. (A) is a figure which shows notionally the direction of the rolling readout on the light-receiving surface 41 in the modification of this invention, (b) shows the exposure timing in each pixel row 46 on the light-receiving surface 41 corresponding to it. FIG. (A) is a figure which shows notionally the direction of the rolling readout on the light-receiving surface 41 in a comparative example, (b) is a figure which shows the exposure timing in each pixel row 46 on the light-receiving surface 41 corresponding to it. is there. (A) is a figure which shows notionally the direction of the rolling readout on the light-receiving surface 41 in a prior art example, (b) is a figure which shows the exposure timing in each pixel row 46 on the light-receiving surface 41 corresponding to it. is there. (A) is a figure which shows notionally the direction of the rolling readout on the light-receiving surface 41 in a comparative example, (b) is a figure which shows the exposure timing in each pixel row 46 on the light-receiving surface 41 corresponding to it. is there.

  Hereinafter, embodiments of a light observation apparatus according to the present invention and an imaging apparatus used therefor will be described in detail with reference to the attached drawings. In the description of the drawings, the same elements will be denoted by the same reference symbols, without redundant description. Also, each drawing is prepared for the purpose of explanation, and is drawn so as to particularly emphasize the target part of the explanation. Therefore, the dimensional ratio of each member in the drawings does not necessarily coincide with the actual one.

FIG. 1 is a schematic block diagram of a light observation apparatus 1 according to an embodiment of the present invention. The light observation device 1 according to the present embodiment is a device that divides the observation light of the object to be observed A into wavelength components, and captures and outputs light images of the divided two wavelength components. As shown in the figure, the light observation apparatus 1 includes a stage 2 on which an observation object A is mounted, a light source 3 for emitting illumination light I to be irradiated to the observation object A, and illumination light I emitted from the light source 3 a collimator lens 4 for condensing the fluorescence to be not occur from the observation object a in accordance with the irradiation of the illumination light I, an objective lens 5 for condensing the observation light B ₁ such reflected light, the illumination light I from the light source 3 the thereby reflected toward the observation object a on the stage 2, a beam splitter 6 which transmits the observation light B ₁ from the observation object a, as two optical image by dividing the observation light B ₁ into two A control unit 9 for controlling the operation of the light splitting optical device 7 for outputting, an imaging device (imaging element) 8 disposed at the imaging position of two light images split by the light splitting optical device 7, and And including. An aperture stop 10, a collimator lens 11, a light splitting optical system 12, and an imaging lens 13 are incorporated in the light splitting optical device 7.

  FIG. 2 is a perspective view showing an example of the configuration of the light splitting optical device 7. As shown in the figure, the light dividing optical device 7 includes a collimator lens 11, an imaging lens 13, a front stage dichroic mirror 25a, a rear stage dichroic mirror 25b, a front stage mirror 26a, a rear stage mirror 26b, and A light dividing optical system 12 including a correction lens 27 is incorporated. Furthermore, an aperture stop 10 having a circular opening is provided on the end face on one end side of the housing 22. The aperture stop 10 has an aperture adjustment mechanism 15 capable of variably setting the inner diameter of the aperture. Hereinafter, the configuration of each component of the light splitting optical device 7 will be described.

The light splitting optical device 7, the observation light B ₁ transmitted through the beam splitter 6 is disposed so as to be incident on the inside of the aperture stop 10. Then, the observation light B _{1 that} has passed through the aperture stop 10 is converted into parallel light by the collimator lens 11 and output along the optical axis L ₁ of the collimator lens 11.

The front stage dichroic mirror 25 a and the rear stage dichroic mirror 25 b are detachably disposed on the optical axis L ₁ inside the housing 22. That is, these dichroic mirrors 25 a and 25 b are integrated and configured to be removable from the optical path of the observation light B ₁ in the housing 22. The dichroic mirrors 25a and 25b may be integrated with the front end mirror 26a, the rear end mirror 26b, and the correction lens 27, which will be described later, and may be detachable. The front dichroic mirror 25a is then disperses the parallel light output from the collimator lens 11, and transmits the first split light B ₂ split into a direction along the optical axis L _1. At the same time, front dichroic mirror 25a is then dispersing the collimated light, it is reflected in a direction perpendicular to the second split light B ₃ spectrally respect to the optical axis L _1.

The post-stage dichroic mirror 25 b further transmits the first divided light B ₂ transmitted through the front-stage dichroic mirror 25 a to form an imaging lens 13 in which the first divided light B ₂ is disposed on the other end side of the housing 22. Output toward At the same time, subsequent dichroic mirror 25b is front mirror 26a after being reflected by the front dichroic mirror 25a, reflects the correction lens 27, and the second split light B ₃ having passed through the subsequent mirror 26b, the second split light Output B ₃ toward the imaging lens 13.

Front mirror 26a reflects the second split beam B ₃ outputted from the pre-dichroic mirror 25a in a direction parallel to the optical axis L _1. Subsequent mirror 26b is reflected toward the light receiving surface of the subsequent dichroic mirror 25b along a direction intersecting the second split light B ₃ having passed through the front mirror 26a to the optical axis L _1. Correcting lens 27 is disposed between the front mirror 26a and the rear mirror 26b, and has a function of performing magnification correction and color correction of the second split light B _3.

An imaging lens 13, the optical axis L ₂ is supported so as to be parallel to the optical axis L _1, perpendicular to the optical axis L ₁ while maintaining a parallel state optical axis L ₂ is the optical axis L ₁ It is configured to be movable in any direction. The imaging lens 13 receives the first and second split lights B ₂ and B ₃ split from the observation light B ₁ via the dichroic mirrors 25 a and 25 b and splits the split lights B ₂ and B ₃ . The image is formed as the first and second separated light images on the imaging device 8 in the camera device 30 disposed outside. At this time, the first and second light images are captured by appropriately adjusting the inner diameter of the aperture stop 10, the angle and position of the rear stage dichroic mirror 25b or the rear stage mirror 26b, and the position of the imaging lens 13. The light can be received by the two divided areas of the light receiving surface of the device 8. On the other hand, the imaging lens 13, the dichroic mirror 25a, when 25b is removed, the observation light B ₁ receives only via the collimator lens 11, the observation light B ₁ of the camera device imaging device 30 8 to form a single light image. At this time, by appropriately adjusting the inner diameter of the aperture stop 10 and the position of the imaging lens 13, it is possible to receive a single light image in a wider area of the entire light receiving surface of the imaging device 8.

The light dividing optical device 7 is provided with a detection mechanism 14 such as a switch element or a sensor element that detects that the front stage dichroic mirror 25a and the rear stage dichroic mirror 25b have been attached and detached, and outputs the detection signal to the control unit 9. It may be done. The dichroic mirrors 25a, may be arranged a wavelength filter such as the first and second band-pass filter on the optical path of the split light B _2, B ₃ divided from the observation light B ₁ via 25b.

  The light dividing optical device 7 having such a configuration has an observation mode in which observation light is observed as a single light image by the camera device 30 (hereinafter referred to as “single view mode”) and observation light by the camera device 30. It can be used both in the observation mode (hereinafter referred to as “double view mode”) in which the two light images are separated and observed.

  Next, the detailed configuration of the imaging device 8 will be described with reference to FIG. As shown in the figure, the imaging device 8 has a plurality of pixel circuits 42 two-dimensionally arranged along the light receiving surface 41 thereof. The pixel circuit 42 includes a photodiode 43 that converts light to electric charge, an amplifier 44 that converts electric charge stored in the photodiode 43 to an electric signal, and a switch 45 that defines the read timing of the electric signal output from the amplifier. It is composed of A plurality of such pixel circuits 42 are arranged at predetermined intervals in one direction (left and right direction in FIG. 3) along the light receiving surface 41 to form a pixel column 46. A plurality of pixel columns 46 are arranged in a direction (vertical direction in FIG. 3) perpendicular to one direction along the light receiving surface 41 to form two adjacent pixel column groups 47a and 47b. That is, of the plurality of pixel columns 46 arranged across the central portion of the light receiving surface 41, the two pixel column groups 47a and 47b are respectively in the regions 48a and 48b obtained by dividing the light receiving surface 41 into two at the central portion. A pixel column 46 is included. The regions 48a and 48b of the light receiving surface 41 correspond to the imaging positions of the first and second light images output from the light splitting optical device 7, respectively.

  Furthermore, the imaging device 8 separately has a circuit configuration capable of rolling readout for each of two pixel column groups 47a and 47b. Specifically, a series circuit including a CDS amplifier 49a and an A / D converter 50a is connected to the output of the amplifier 44 of each pixel circuit 42 included in the region 48a. The number of the pixel circuits 42 is equal to the number of the arrayed pixel circuits 42 and commonly connected along the arrangement direction of the pixel columns 46 across the plurality of pixel columns 46 included in the pixel column group 47a. A digital signal output circuit (signal readout circuit) 51a is commonly connected to these series circuits. With such a configuration, the detection signal of the light image detected by the pixel circuit 42 constituting the pixel column 46 included in the pixel column group 47a is sequentially read by the digital signal output circuit 51a as a digital signal for each pixel column 46. .

  Similarly, a series circuit including a CDS amplifier 49b and an A / D converter 50b is connected to the output of the amplifier 44 of each pixel circuit 42 included in the region 48b, and these series circuits are included in the pixel column group 47b. The plurality of pixel columns 46 are commonly connected along the arrangement direction of the pixel columns 46. A digital signal output circuit (signal readout circuit) 51b is commonly connected to these series circuits. With such a configuration, the detection signal of the light image detected by the pixel circuit 42 constituting the pixel column 46 included in the pixel column group 47 b is sequentially read by the digital signal output circuit 51 b as a digital signal for each pixel column 46 .

  Further, the imaging device 8 is further provided with a scanning circuit 52 that defines the exposure timing and the signal readout timing in the pixel circuit 42. The scanning circuit 52 defines the read timing of the electric signal by the switch 45 of the pixel circuit 42 for each pixel column 46. That is, the scanning circuit 52 defines the readout timing so that the readout of the electrical signal from the pixel circuit 42 is performed in the order of a predetermined direction along the light receiving surface 41 for each pixel column 46, so-called rolling readout. . Here, the scanning circuit 52 defines readout timing so that rolling readout is performed independently for the pixel array group 47a included in the area 48a and the pixel array group 47b included in the area 48b. Furthermore, in response to the instruction signal from control unit 9, scanning circuit 52 performs rolling along light receiving surface 41 independently for pixel column group 47a included in area 48a and pixel column group 47b included in area 48b. It is configured to be able to set the direction of reading.

Returning to FIG. 1, the control unit 9 is connected to the imaging device 8, and the rolling readout direction from the pixel column group 47 a in the imaging device 8 and the rolling readout direction from the pixel column group 47 b in the imaging device 8 Generates an instruction signal to independently control the The control unit 9 may be a control device such as a computer terminal connected to the outside of the camera device 30 or may be a control circuit mounted on the imaging device 8. Specifically, when the light dividing optical device 7 is set to the double view mode, the control unit 9 specifically includes the light dividing optical system 12 (FIG. 2) including the front dichroic mirror 25 a and the rear dichroic mirror 25 b. Is disposed on the light path of the observation light B ₁ , the following control is performed. Specifically, the direction of the rolling readout in the pixel column group 47a and the method of the rolling readout in the pixel column group 47b are controlled to be the same direction in the parallel direction of the pixel columns 46. FIG. 4 is a diagram conceptually showing the direction of the rolling readout in the pixel array groups 47 a and 47 b on the light receiving surface 41 controlled by the control unit 9. Thus, in the pixel column group 47a, the direction of rolling readout is controlled from the end opposite to the area 48b of the area 48a in the direction toward the boundary between the area 48a and the area 48b (downward in FIG. 4) In the pixel column group 47b, the direction of the rolling readout is controlled from the boundary between the area 48b and the area 48a in the direction (downward in FIG. 4) directed to the end opposite to the area 48a of the area 48b. .

  In addition, when the light dividing optical device 7 is set to the single view mode, the control unit 9 is an observation light of the light dividing optical system 12 (FIG. 2) including the front stage dichroic mirror 25a and the rear stage dichroic mirror 25b. When removed from the optical path of B1 and separated, control is performed as follows. Specifically, control is performed so that the rolling readout direction in the pixel column group 47a and the rolling readout method in the pixel column group 47b are opposite in the parallel direction of the pixel columns 46. FIG. 5 is a diagram conceptually showing the direction of the rolling readout in the pixel array groups 47a and 47b on the light receiving surface 41 controlled by the control unit 9. As shown in FIG. Thus, in the pixel column group 47a, the direction of rolling readout is controlled from the boundary between the area 48a and the area 48b in the direction (upward in FIG. 4) from the boundary between the area 48a and the area 48b of the area 48a. In the pixel column group 47b, the direction of the rolling readout is controlled from the boundary between the area 48b and the area 48a in the direction (downward in FIG. 4) directed to the end opposite to the area 48a of the area 48b. .

  Here, when the light dividing optical device 7 is provided with a detection mechanism for detecting that the front stage dichroic mirror 25a and the rear stage dichroic mirror 25b are attached and detached, the control unit 9 switches the reading mode as follows. You may control as follows. That is, when it is detected that the light dividing optical system 12 including the front stage dichroic mirror 25a and the rear stage dichroic mirror 25b is disposed on the optical path, the control unit 9 detects the pixel array group 47a as shown in FIG. , 47b are controlled so as to automatically switch to the first readout mode in which the direction of the rolling readout is the same. On the other hand, when it is detected that the light dividing optical system 12 is separated from the light path, the control unit 9 reverses the rolling readout direction in the pixel row groups 47a and 47b as shown in FIG. Control is made to automatically switch to the second read mode. When the light splitting optical device 7 is not provided with a detection mechanism, the control unit 9 may switch the reading mode according to the selection of the observer.

  Next, the procedure of the light observation method according to the present embodiment using the light observation device 1 will be described in detail. FIG. 6 is a flowchart showing the procedure of the light observation method.

When the observation process is started, first, the observer selects the observation mode (S0) to determine the rolling readout direction. Specifically, the observer uses the selection screen displayed on the display unit of the control unit 9 to select the single view mode or the double view mode. For double view mode, the observation light _{B 1} represents, is divided by the light splitting optical system 12 to the first split light _{B 2} and the second split light _{B 3} (S11). Then, first divided light B ₂ and the second split light B ₃ is imaged, the second corresponding to the first light image and the second split light B ₃ corresponding to the first split light B ₂ Light image is formed (S21). The imaging device 8 is disposed such that the pixel array group 47a is located at the position where the first light image is formed, and the pixel array group 47b is located at the position where the second light image is formed. In the case of the double view mode, the first readout mode in which the rolling readout direction is the same in the pixel array group 47a and the pixel array group 47b is selected, and the first light image and the second light image are the first. The image is captured in the read mode (S31). Based on the obtained image data, a first image corresponding to the first light image and a second image corresponding to the second light image are created and displayed on the display unit of the control unit 9 ( S4).

On the other hand, if the single view mode is selected, the observation light B ₁ is imaged optical image is formed (S22). In the single view mode, the second readout mode in which the rolling readout direction is reverse is selected by the pixel array group 47a and the pixel array group 47b, and the light image is captured in the second readout mode (S32) ). Based on the image data obtained by imaging, an image corresponding to the light image is created and displayed on the display unit of the control unit 9 (S4).

  According to the light observation device 1 described above, the imaging device 8 used for that, and the light observation method, the observation light of the observation object A is divided into the first and second division lights by the light division optical system 12 and divided. The first and second divided light beams are imaged by the imaging lens 13 to form first and second light images, and the first light image is formed into the pixel row group 47 a included in the area 48 a of the imaging device 8. The second light image is received by the pixel row group 47 b included in the area 48 b of the imaging device 8. Then, the detection signals of the first and second light images are independently read by rolling from the pixel row groups 47a and 47b under the control of the control unit 9, and the direction of the rolling readout in the pixel row group 47a and the pixel row group The direction of the rolling readout at 47b is made identical. As a result, the exposure timings at the same site of the observation object A can be easily aligned between the detection image of the first light image and the detection image of the second light image, so the exposure condition of each site of the two light images Can be made uniform. As a result, comparative observation using detected images of two light images can be facilitated.

FIG. 7A conceptually shows the direction of the rolling readout on the light receiving surface 41 controlled by the control unit 9 when used in the double view mode, and correspondingly, in FIG. The exposure timing in each pixel row 46 on the light receiving surface 41 when used in the double view mode is shown. As shown in the figure, by the direction of rolling read in the region 48a and the region 48b is set to the same, the exposure period T _A1 of the plurality of pixel columns 46 contained in the region 48a is, from the end regions 48a is set so as to sequentially delayed by a predetermined time period toward the center portion of the light-receiving surface 41, the exposure period T _B1 of a plurality of pixel columns 46 contained in the area 48b is, toward the end of the region 48b from the central portion of the light-receiving surface 41 Are set to be sequentially delayed by a predetermined period. As a result, the exposure periods T _A1 and T _{B1 of} the pixel row 46 corresponding to the same portion of the detected image are easily matched between the regions 48 a and 48 b. Thereby, when the observation object A moves or when the observation light changes due to deterioration of the fluorescent reagent etc., the detected image of the first light image and the detected image of the second light image are accurately compared. Can.

  Here, the control unit 9 is capable of switching the read mode between the first read mode and the second read mode. The digital signal output circuits 51a and 51b and the scanning circuit 52 operate to select a readout mode from the first readout mode and the second readout mode under the control of the control unit 9. This makes it possible to equalize the exposure conditions of each part of the two light images when used in the double view mode. On the other hand, when used in the single view mode, it is possible to prevent the occurrence of linear contrast in the detection image of a single light image. As a result, even when the single view mode and the double view mode are shared, it is possible to observe the light image with high accuracy.

FIG. 8A conceptually shows the direction of the rolling readout on the light receiving surface 41 controlled by the control unit 9 when used in the single view mode, and correspondingly, in FIG. The exposure timing in each pixel row 46 on the light receiving surface 41 at the time of use in the single view mode is shown. As shown in the figure, by setting the rolling readout directions in the regions 48 a and 48 b in opposite directions, the exposure period TA ₂ of the plurality of pixel rows 46 included in the region 48 a corresponds to the central portion of the light receiving surface 41. towards the end of the area 48a from being set to delayed successively by a predetermined time period, the exposure period T _B2 of the plurality of pixel columns 46 contained in the area 48b is, toward the end of the region 48b from the central portion of the light-receiving surface 41 Are set to be sequentially delayed by a predetermined period. As a result, the exposure periods T _A2 and T _{B2 of} the pixel row 46 adjacent to the detected image at the central portion easily coincide between the regions 48 a and 48 b. This makes it possible to prevent the occurrence of linear contrast between the area 48a and the area 48b in the single light image detection image.

On the other hand, in FIG. 12, each pixel row 46 on the light receiving surface 41 in the case of the first readout mode in which the rolling readout directions in the pixel column groups 47 a and 47 b are the same direction when using the single view mode. Shows the exposure timing in With this setting, the exposure period T _A3, T _B3 of adjacent two pixel columns 46 corresponding to the central portion of the detection image, regions 48a, undesirably becomes inconsistent between 48b. In this case, a linear light-dark difference at the boundary between the area 48a and the area 48b is likely to occur in the detected image of a single light image.

Further, FIG. 13 shows the exposure timing in each pixel row 46 on the light receiving surface 41 in the case where a conventional rolling readout type imaging device is used. In such a case, by the rolling readout is performed in one direction across the light receiving surface 41, the exposure period T ₄ of a plurality of pixel columns 46, the predetermined time period from the one end toward the other end of the light-receiving surface 41 Are set to be delayed in sequence. As a result, the exposure period T ₄ of the pixel row 46 corresponding to the same site of the two detection images when used in a double view mode tends to mismatch. As a result, when the observation object A moves or when the observation light changes due to deterioration of the fluorescent reagent, etc., the exposure conditions of the detected image of the first light image and the detected image of the second light image differ, You can not compare them correctly.

Further, FIG. 14 shows the exposure timing in each pixel row 46 on the light receiving surface 41 when setting in the second readout mode when using the double view mode. In this case, the exposure period _T A5 of the pixel row 46 corresponding to the same site of the detection _{image, T B5} are region 48a, it becomes mismatched between 48b. As a result, the exposure conditions of the detected image of the first light image and the detected image of the second light image are different, and they can not be compared accurately.

  The present invention is not limited to the embodiments described above.

  For example, the first and second readout modes set by the control unit 9 of the light observation device 1 can be changed as follows.

  FIGS. 9 and 10 are diagrams conceptually showing the direction of rolling readout in the pixel array groups 47a and 47b on the light receiving surface 41 controlled by the controller 9 according to the modification of the present invention. As shown in FIG. 9, when the double view mode is used, the rolling readout directions of the pixel column groups 47a and 47b may be set opposite to the direction shown in FIG. Further, as shown in FIG. 10, when the single view mode is used, the rolling readout directions in the pixel column groups 47a and 47b may be set opposite to the direction shown in FIG.

In addition, the control unit 9 may be configured to be capable of independently changing the exposure time for each pixel row group 47a, 47b, and is configured to be able to control the exposure time for each pixel row group 47a, 47b to be different. It may be done. In FIG. 11, the exposure timing in each pixel row 46 on the light receiving surface 41 in the modification of the present invention is shown. As shown in the figure, the control unit 9, the pixel row group 47a, to control the direction of rolling readout for each 47b, each pixel row group 47a, the length of the 48b of the exposure period _{T A6, T B6} different The time is configurable. In this case, the start timing of the signal readout of each pixel column 46 is set immediately after the exposure period _TA6 , _TB6 of each pixel column, and the start timing of the signal readout is controlled to be synchronized between the respective regions 48a, 48b. Ru. That is, the timings of signal readout of the two pixel columns 46 in the regions 48a and 48b corresponding to the same portion of the detected image are controlled to be synchronized with each other.

  In the conventional imaging of observation light in the double view mode, optical images of different wavelength regions are imaged respectively, but when the light intensities of the respective optical images are greatly different, a light reduction filter etc. I am passing In the present embodiment shown in FIG. 11, since the exposure time can be set independently between the two light images, the exposure time of one light image having the larger light intensity is made shorter than the other. The light intensity detected as a detection image can be made comparable even without using a light reduction filter. Furthermore, by synchronizing the timing of signal readout of the two pixel columns 46 in the two regions 48a and 48b, the exposure timings at the same part of the observation object are surely aligned between the detected images of the two light images. It is possible to make the exposure conditions of each part of the two light images more uniform.

  Further, the configuration of the light dividing optical device 7 constituting the light observation device 1 is not limited to the configuration shown in FIG. 2 and may be, for example, US Pat. No. 5,926,283 or US Pat. No. 7,667,761 The light-splitting optical system described in “K. Kinosita, Jr. et al.,“ DualView Microscopy with a Single Camera: Real-Time Imaging of Molecular Orientations and Calcium ”, The Journal of Cell Biology, Volume 115, Number 1, The conventional configuration such as the optical system described in October 1991, pp 67-73 may be used. Further, the light splitting optical system 12 is not limited to the configuration including the dichroic mirror.

  A light observation apparatus according to an embodiment of the present invention includes a light dividing optical system that receives observation light of an object from the outside and divides the observation light into first and second lights, and first and second lights. An imaging lens for imaging the first and second light to form a first and second light image, and an imaging position of the first and second light image, A plurality of pixel columns including a plurality of pixels are arranged side by side, and a first pixel column group among a plurality of pixel columns included in a first region corresponding to an imaging position of a first light image; An imaging element capable of independently performing rolling readout in a second pixel row group among a plurality of pixel rows included in a second region corresponding to the imaging position of the light image, and included in the first region A control unit that controls rolling readout in the first pixel column group and the second pixel column group included in the second region; Unit may include, by way of the rolling readout in the first pixel row group, and the direction of the rolling readout in the second pixel row group is controlled to be the same in the parallel direction of the plurality of pixel columns.

  Alternatively, in the light observation method according to another embodiment of the present invention, a plurality of first pixel column groups, each including a plurality of pixel columns including a plurality of pixels, are arranged adjacent to the first pixel column group. An imaging device having a second pixel column group in which a plurality of pixel columns including pixels are arranged side by side, and capable of performing rolling readout of the first pixel column group and the second pixel column group In the light observation method, the observation light of the object is divided into the first and second lights from the outside, the first light is imaged, and the first light image is made incident on the first pixel row group Forming a second light image to form a second light image so as to be incident on the second group of pixel columns, the direction of rolling readout in the first group of pixel columns, and The direction of the rolling readout in the group of pixel columns is controlled to be the same in the parallel direction of the plurality of pixel columns.

  According to such a light observation apparatus or light observation method, the observation light of the object is divided into the first and second lights, and the divided first and second lights are imaged to form the first and second lights. A second light image is formed, the first light image is received by the first pixel column group included in the first region of the imaging device, and the second light image is included in the second region of the imaging device Light is received by the second pixel column group. Then, the detection signals of the first and second light images are read by rolling independently from the first and second pixel column groups, respectively, and the direction of the rolling readout in the first pixel column group and the second pixel column The direction of the rolling readout in the group is made identical. This makes it easy to align the exposure timing on the same part of the object between the detection image of the first light image and the detection image of the second light image, so the exposure conditions of each part of the two light images are uniform. Can be As a result, comparative observation using detected images of two light images can be facilitated.

  Here, the control unit is configured such that the direction of the rolling readout in the first pixel column group and the direction of the rolling readout in the second pixel column group are the same in the parallel direction of the plurality of pixel columns. The second readout mode in which the readout mode, the rolling readout direction in the first pixel column group, and the rolling readout direction in the second pixel column group are opposite in the parallel direction of the plurality of pixel columns It is preferable to control to be switchable. In addition, a first readout mode in which the rolling readout direction in the first pixel column group and the rolling readout direction in the second pixel column group are the same in the parallel direction of the plurality of pixel columns; The second readout mode is selected such that the rolling readout direction in one pixel column group and the rolling readout direction in the second pixel column group are opposite in the parallel direction of the plurality of pixel columns. Is preferred. With this configuration, when two light images are simultaneously captured by the imaging device, the exposure condition of each portion of the two light images can be made uniform by switching to the first readout mode. On the other hand, when a single light image is captured by the imaging device, switching to the second readout mode can prevent the occurrence of linear contrast in the detection image of the single light image. As a result, even when the observation mode for capturing two light images and the observation mode for capturing a single light image are shared, it is possible to observe the light image with high accuracy.

  The light splitting optical system is configured to be removable from the light path of the observation light, and the control unit switches to the first readout mode when the light splitting optical system is disposed on the light path. It is also preferable to control to switch to the second readout mode when the light splitting optical system is separated from the light path. In this case, since the first and second readout modes are controlled to be switched according to attachment and detachment of the light splitting optical system, an observation mode for imaging two light images and an observation mode for imaging a single light image Makes it easy to observe the light image with high accuracy.

  Furthermore, it is preferable that the control unit variably controls the exposure time for rolling readout in the first pixel column group and the exposure time for rolling readout in the second pixel column group. By so doing, the sensitivity for capturing the first light image and the sensitivity for capturing the second light image can be set freely, and the degree of freedom of the imaging conditions for the two light images can be increased.

  Furthermore, it is preferable that the control unit controls the exposure time for rolling readout in the first pixel column group and the exposure time for rolling readout in the second pixel column group to different times. By so doing, the sensitivity for capturing the first light image and the sensitivity for capturing the second light image can be set to be different, and the degree of freedom of the imaging conditions for the two light images can be increased.

  Furthermore, it is preferable that the control unit performs control such that the start timing of the rolling readout in the first pixel column group and the start timing of the rolling readout in the second pixel column group are synchronized. According to such a configuration, it is possible to reliably align the exposure timing at the same portion of the object between the detection image of the first light image and the detection image of the second light image, and two light images It is possible to further uniform the exposure conditions of each part of.

  Furthermore, the imaging apparatus according to the embodiment of the present invention divides the observation light of the object from the outside into the first and second lights, and generates the first and second lights generated based on the first and second lights. An imaging apparatus for use in a light observation apparatus for simultaneously imaging two light images, the first pixel column group including a plurality of pixel columns including a plurality of pixels arranged adjacent to the first pixel column group A second pixel column group in which a plurality of pixel columns including a plurality of pixels are arranged side by side, a first signal readout circuit for processing signal readout from the first pixel column group, and a second pixel column And a second signal readout circuit that processes signal readout from the group independently of the first signal readout circuit, and the first signal readout circuit performs signal readout in the first pixel column group by rolling readout. Processed and rolling read in multiple pixel sequences The second signal readout circuit processes the signal readout in the second pixel column group by rolling readout, and the direction of the rolling readout is a parallel direction of a plurality of pixel columns. Set to one direction in

  According to this imaging device, the first light image formed on the basis of the first light divided from the observation light of the object is divided by the first pixel row group included in the first region of the imaging device. It is possible to receive light. At the same time, a second light image formed on the basis of the second light divided from the observation light of the object is made receivable by the second group of pixel columns included in the second region of the imaging device. Ru. Then, the detection signals of the first and second light images are read by rolling independently from the first and second pixel column groups by the first and second signal readout circuits, respectively, and the first pixel column group is read. The direction of the rolling readout in the second pixel row and the direction of the rolling readout in the second pixel row are made the same. This makes it easy to align the exposure timing on the same part of the object between the detection image of the first light image and the detection image of the second light image, so the exposure conditions of each part of the two light images are uniform. Can be As a result, comparative observation using detected images of two light images can be facilitated.

  Here, in the first and second signal readout circuits, the rolling readout direction in the first pixel column group and the rolling readout direction in the second pixel column group are the same as those of the plurality of pixel columns. The first readout mode, which is the same in the parallel direction, the rolling readout direction in the first pixel column group, and the rolling readout direction in the second pixel column group are the same as in the plurality of pixel columns. It is also preferable to switch between the second read mode in which the parallel direction is opposite to the second read mode. With this configuration, when two light images are simultaneously captured by the imaging device, the exposure condition of each portion of the two light images can be made uniform by switching to the first readout mode. On the other hand, when a single light image is captured by the imaging device, switching to the second readout mode can prevent the occurrence of linear contrast in the detection image of the single light image. As a result, even when the observation mode for capturing two light images and the observation mode for capturing a single light image are shared, it is possible to observe the light image with high accuracy.

DESCRIPTION OF SYMBOLS 1 ... light observation apparatus, 7 ... light division optical apparatus, 8 ... imaging apparatus, 9 ... control part, 12 ... light division optical system, 13 ... imaging lens, 42 ... pixel circuit, 46 ... each pixel row, 47a ... 47th 1 pixel row group 47b second pixel row group 48a first region 48b second region 51a digital signal output circuit (first signal readout circuit) 51b digital signal output circuit (Second signal readout circuit) A: observation object B ₁ : observation light B ₂ : first split light B ₃ : second split light

Claims (9)

A light dividing optical element that receives observation light of an object from the outside and divides the observation light into first and second lights ;
A first pixel column group in which a plurality of pixel columns including a plurality of pixels are arranged side by side, and a second pixel in which a plurality of pixel columns including a plurality of pixels are arranged side by side adjacent to the first pixel column group. A pixel column group, a first signal readout circuit that processes signal readout from the first pixel column group, and signal readout from the second pixel column group independently of the first signal readout circuit An imaging device having a second signal readout circuit for processing ;
A control unit that performs rolling readout control by the imaging device ;
The control unit is configured such that the rolling readout direction in the first pixel column group and the rolling readout direction in the second pixel column group are the same in the parallel direction of the plurality of pixel columns. A readout mode of the first pixel column group, a rolling readout direction of the first pixel column group, and a rolling readout direction of the second pixel column group in opposite directions in the parallel direction of the plurality of pixel columns; Controlling the rolling readout by the imaging device according to one of the two readout modes;
When the light splitting optical element is disposed on the light path of the observation light, the first readout mode is selected, and when the light splitting optical element is separated from the light path The second read mode is selected,
Light observation device . The control unit variably controls an exposure time of rolling readout in the first pixel column group and an exposure time of rolling readout in the second pixel column group.
Optical observation device according to 請 Motomeko 1. The control unit controls the exposure time for rolling readout in the first pixel column group and the exposure time for rolling readout in the second pixel column group to different times.
Optical observation device 請 Motomeko 1 or 2, wherein.   It further comprises a detection mechanism for detecting the attachment / detachment of the light dividing optical element,
  The control unit according to any one of claims 1 to 3, wherein one of the first read mode and the second read mode is selected based on the detection result of the detection mechanism. Light observation device.   A light dividing optical element that receives observation light of an object from the outside and divides the observation light into first and second lights, and a first pixel in which a plurality of pixel columns including a plurality of pixels are arranged side by side A first group of pixel columns and a second group of pixel columns adjacent to the first group of pixel columns and including a plurality of pixel columns including a plurality of pixels arranged side by side; In a light observation method using an imaging device capable of performing rolling readout with each of two pixel column groups,
  A first readout mode in which the rolling readout direction in the first pixel column group and the rolling readout direction in the second pixel column group are the same in the parallel direction of the plurality of pixel columns; In a second readout mode in which the rolling readout direction in the first pixel column group and the rolling readout direction in the second pixel column group are opposite in the parallel direction of the plurality of pixel columns A selection step of selecting one of the read modes;
  An imaging step of controlling rolling readout by the imaging device based on the readout mode selected in the selection step;
  In the selection step, when the light splitting optical element is disposed on the light path of the observation light, the first readout mode is selected, and the light splitting optical element is separated from the light path. At the time, the second read mode is selected,
Light observation method.   In the imaging step, an exposure time of rolling readout in the first pixel column group and an exposure time of rolling readout in the second pixel column group are variably controlled.
The light observation method according to claim 5.   In the imaging step, the exposure time for rolling readout in the first pixel column group and the exposure time for rolling readout in the second pixel column group are controlled to different times.
A light observation method according to claim 5 or 6.   The method further comprises a detection step of detecting attachment / detachment of the light dividing optical element,
  In the selection step, one of the first read mode and the second read mode is selected based on the detection result of the detection step.
The light observation method according to any one of claims 5 to 7.   In the selection step, one of the first readout mode and the second readout mode is selected in response to the selection input of the observer.
The light observation method according to any one of claims 5 to 7.

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