JP2007003323A - Photographing system - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To acquire an image free from an influence of a magnification chromatic aberration of a lens, when picking up images from the same subject by different wavelengths of fluorescence. <P>SOLUTION: This photographing system 1 provided with a photographing device 10 for photographing the fluorescent image of the subject, and a photographing controller 100 for controlling the photographing device 10, is provided with a chromatic aberration correcting means for moving at least one of an imaging element 31 and the subject 5 along an optical axis direction of an image-focusing lens 32 to correct the magnification chromatic aberration of the image-focusing lens 32 generated by a wavelength difference between the first wavelength and the second wavelength, before photographing the fluorescent image of the second wavelength different from the first wavelength, after photographing the fluorescent image of the first wavelength. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a photographing system that captures a fluorescent image of a subject arranged in a casing.

  2. Description of the Related Art Conventionally, an apparatus that images a subject by arranging the subject in a casing and irradiating the subject with a light source provided in the casing is used in various fields. For example, in the field of biochemistry, a fluorescence detection system using a fluorescent substance as a labeling substance is known. According to this fluorescence detection system, gene sequences, gene expression levels, protein separation, identification, molecular weight, and property evaluation can be performed by reading fluorescence images.

  Specifically, for example, after adding a fluorescent dye into a solution containing a plurality of DNA fragments to be electrophoresed, the plurality of DNA fragments are electrophoresed on a gel support. Alternatively, a plurality of DNA fragments are electrophoresed on a gel support containing a fluorescent dye. The plurality of DNA fragments are electrophoresed on a gel support, and the electrophoretic DNA fragments are labeled by, for example, immersing the gel support in a solution containing a fluorescent dye. By exciting the fluorescent dye and detecting the resulting fluorescence, an image can be generated and the DNA distribution on the gel support can be detected.

  Alternatively, after electrophoresis of a plurality of DNA fragments on a gel support, the DNA is denatured, and then at least a part of the denatured DNA fragments is transferred onto a transfer support such as nitrocellulose by Southern blotting. A probe prepared by transcription and labeling a DNA or RNA complementary to the target DNA with a fluorescent dye is hybridized with a denatured DNA fragment, and only the DNA fragment complementary to the probe DNA or probe RNA is selectively selected. By labeling, exciting the fluorescent dye with excitation light, and detecting the resulting fluorescence, an image can be generated and the intended DNA distribution on the transfer support can be detected.

  In recent years, microarray analysis systems have attracted attention as biochemical analysis systems. For example, in a microarray analysis system using a fluorescent substance as a labeling substance, hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, etc., at different positions on the carrier surface such as a slide glass plate or membrane filter, Use a spotter device to drop a specific binding substance that can specifically bind to a biological substance such as nucleic acid, cDNA, DNA, or RNA, and whose base sequence, base length, or composition is known. A large number of independent spots, and then extracted from the living body by extraction, isolation, etc. of hormones, tumor markers, enzymes, antibodies, antigens, abzymes, other proteins, nucleic acids, cDNA, DNA, mRNA, etc. Or a biologically-derived substance that has been subjected to chemical or chemical modification treatment, and a fluorescent label such as a fluorescent dye Fluorescence emitted from fluorescent substances, dyes, and other labeling substances is emitted by irradiating microarrays that are specifically bound to the specific binding substances to the specific binding substances by hybridization, etc. A substance derived from a living body can be analyzed by photoelectric detection.

  In addition to the fluorescence method described above, a reaction (chemiluminescence) that emits light when a substance undergoes a chemical reaction is used to examine and study biological tissues (gene analysis, disease, aging), organic It has been proposed to evaluate deterioration of compounds and polymer compounds (chemiluminescence method). In this chemiluminescence method, for example, a substance derived from a living body is labeled with an antigen, and an antibody labeled with a labeling substance that generates chemiluminescence is brought into contact with the chemiluminescent substrate. Chemoluminescence is generated by contact.

  In the above-described fluorescence method or chemiluminescence method, the fluorescence or luminescence to be detected is weak light. Therefore, when detecting fluorescence or luminescence, the subject is housed in a casing that blocks external light and emitted from the subject. Fluorescence or light emission is detected by a cooled CCD (imaging unit) (for example, see Patent Document 1). Specifically, in the fluorescence method, a subject is irradiated with light from an excitation light source provided in a housing, and the fluorescence emitted from the subject is imaged and visualized on a cooled CCD through a high sensitivity lens. With the excitation light source turned off, the light emitted from the subject is imaged on a cooled CCD through a high sensitivity lens and visualized. Patent Document 1 discloses a subject moving unit for easily displacing the imaging distance and a movement control unit for controlling the subject moving unit in order to capture an image with a desired size even if the size of the subject changes. An imaging system provided is disclosed.

Furthermore, in the biochemical analysis system as described above, substances labeled with phosphors that generate fluorescence of different wavelengths are distributed on one subject, fluorescent images of different wavelengths are taken, and the images are superimposed. Systems that can be displayed together have been proposed.
JP 2003-287497 A

  However, when fluorescent images of different wavelengths are taken and displayed in an overlapping manner, there is a problem in that the size of the projected image in the acquired fluorescent image differs if the fluorescent wavelength is different due to the influence of lateral chromatic aberration of the imaging lens. Arise.

  As a means for solving this problem, a method is conceivable in which image processing for enlarging or reducing the acquired fluorescent images of different wavelengths is performed, and the sizes of the projected images among a plurality of fluorescent images are matched. However, such image processing is not desirable because the resolution is lowered and the edges of the image are blurred.

  Although it is conceivable to provide a telecentric lens as the imaging lens, it is difficult to realize a bright lens because the F-number increases when the telecentric lens is used. Furthermore, there is a problem that the use of a lens corrected for chromatic aberration increases the cost of the lens system.

  Therefore, the present invention is not affected by the magnification chromatic aberration of the imaging lens between the plurality of fluorescent images without increasing the cost of the imaging lens when photographing a plurality of fluorescent images with different wavelengths from the same subject. An object is to provide a photographing system capable of obtaining an image.

An imaging system according to the present invention includes an imaging apparatus including a housing that accommodates a subject in which a plurality of areas in which a substance labeled with a phosphor is distributed is set, and an imaging unit that captures a fluorescent image of the subject, and In a shooting system equipped with a shooting control device that controls the operation of the shooting unit,
The imaging unit includes an imaging element that acquires a fluorescent image of a subject, and an imaging lens that forms an image of the fluorescent image on an imaging surface of the imaging element;
The imaging control device controls the imaging unit to capture a fluorescent image of a second wavelength different from the first wavelength of the subject after capturing a fluorescent image of the first wavelength of the subject, Obtaining an image obtained by superimposing the fluorescence image of the first wavelength and the fluorescence image of the second wavelength;
After taking the fluorescence image of the first wavelength, before taking the fluorescence image of the second wavelength, the chromatic aberration of magnification of the imaging lens caused by the wavelength difference between the first wavelength and the second wavelength Chromatic aberration correction means for moving at least one of the image sensor and the subject in the optical axis direction of the imaging lens so as to correct the image.

  The imaging system of the present invention stores a table in which the relationship between the chromatic aberration of magnification based on the difference between the first wavelength and the second wavelength and the optimum difference in imaging distance at each wavelength is stored. Preferably, a storage unit is provided, and the chromatic aberration correction unit moves at least one of the image sensor and the subject based on the relationship.

  The relationship held in the table may be obtained based on design information of the imaging lens, or may be obtained by actual measurement before photographing the fluorescence image. .

  The chromatic aberration correcting means preferably includes a pulse motor as means for moving at least one of the image sensor and the subject.

  According to the imaging system of the present invention, after the fluorescent image of the first wavelength is captured and before the fluorescent image of the second wavelength is captured, the result of the wavelength difference between the first wavelength and the second wavelength occurs. In order to correct the chromatic aberration of magnification of the image lens, chromatic aberration correction means for moving at least one of the image sensor and the subject in the optical axis direction of the imaging lens is provided, so that the imaging lens has a telecentric optical system and chromatic aberration correction. Since it is possible to correct chromatic aberration when photographing fluorescent images of different wavelengths without providing an optical system, fluorescent images that are not affected by the chromatic aberration of magnification of the imaging lens can be obtained without increasing the cost of the imaging lens. Obtainable. That is, in the image obtained by superimposing the fluorescence image of the first wavelength and the fluorescence image of the second wavelength, there is no deviation in the projection size between the two fluorescence images, so that an accurate distribution region of substances labeled with different phosphors Can be displayed.

  In addition, the imaging system includes storage means for storing a table in which the relationship between the chromatic aberration of magnification based on the difference between the first wavelength and the second wavelength and the optimum difference in imaging distance at each wavelength is stored. If the chromatic aberration correcting means moves at least one of the image sensor and the subject based on the relationship held in the table, chromatic aberration can be corrected (adjusted) efficiently.

  DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of the photographing apparatus of the present invention will be described below in detail with reference to the drawings. FIG. 1 is a perspective view showing an example of a photographing system using the photographing control apparatus of the present invention. The imaging system 1 in FIG. 1 is a fluorescence detection system that detects fluorescence emitted from a substance and displays a fluorescence image by irradiating excitation light onto a subject on which a plurality of substances labeled with phosphors are distributed, for example. The photographing apparatus 10 and the photographing control apparatus 100 are included. The imaging control device 100 is composed of, for example, a personal computer, and is connected to an input unit 2 such as a mouse and a keyboard and a display unit 3 including a CRT and a liquid crystal display. The photographing apparatus 10 is controlled by the photographing control apparatus 100 to photograph a fluorescent image of the subject 5, sends the acquired image information to the photographing control apparatus 100, and the photographing control apparatus 100 displays the image information on the display unit 3. It has become.

  Here, the subject will be described as one in which a plurality of fluorescently labeled analytes labeled with fluorescent dyes emit fluorescence when irradiated with excitation light (fluorescence method), but in contact with a chemiluminescent substrate, A specimen in which luminescence is distributed may be distributed (chemiluminescence method).

  FIG. 2 is a plan view schematically showing a schematic configuration of the photographing apparatus in the photographing system of the present invention. The photographing apparatus 10 will be described with reference to FIGS. 1 and 2. The photographing apparatus 10 includes a housing 20 that houses the subject 5 and a photographing unit 30 that captures a fluorescent image of the subject 5 housed in the housing 20 and outputs image information.

  Here, the housing 20 has a hollow portion 21 formed in a substantially rectangular parallelepiped, and includes a subject placement portion 40 in which the subject 5 is placed and a subject movement that moves the subject placement portion 40 in the direction of arrow Z. Part 60 is provided. In addition, a lid 22 is attached to the housing 20 so as to be openable and closable so that the user can open and close the lid 22 and put the subject 5 in and out of the housing 20. As described above, the housing 20 forms a dark box that does not allow outside light to enter the hollow portion 21, and photography is possible even when weak light is emitted from the subject.

  The photographing unit 30 captures a fluorescent image of the subject 5 and outputs image information, and includes an imaging unit 31 including a plurality of two-dimensionally arranged imaging elements such as CCDs, and a fluorescent image on the imaging surface 31a of the imaging element. And a lens unit 32 which is an imaging lens for forming an image.

  The imaging unit 31 is fixed to the upper surface 20a of the housing 20, and a cooling unit (not shown) is attached to the imaging unit 31, and the image unit includes a noise component due to dark current by cooling the imaging unit 30. Can be prevented. The lens unit 32 is configured by combining a plurality of lenses, and is provided with an autofocus function for automatically focusing on the subject 5.

  An excitation light cut filter 35 is provided on the subject side of the lens unit 32 so that the excitation light reflected from the subject 5 does not enter the imaging unit.

  The subject placement unit 40 includes a housing 41 and a placement surface 42, and has a structure in which the subject 5 can be placed on the placement surface 42 provided in the housing 41. The subject placement unit 40 is held by the subject moving unit 60 and can be moved in the arrow Z direction by the operation of the subject moving unit 60.

  A light source 50 made of, for example, an LED is fixed to the subject placement unit 40. The light source 50 emits excitation light for causing the phosphor to emit light, and includes a bottom light source 51 and an incident light source 52. The bottom light source 51 is fixed to the housing 41 and is provided below the arrangement surface 42. The epi-illumination light source 52 is fixed to the housing 41 and is provided above the arrangement surface 42. Therefore, the bottom light source 51 emits excitation light from below the subject 5, and the epi-illumination light source 52 emits excitation light from above the subject 5. The light source may be selectively driven according to the subject to be photographed. In the present embodiment, an incident light source 52 that emits excitation light from above the subject 5 is used.

  The incident light source 52 is, for example, one in which a large number of LEDs are two-dimensionally arranged. A plurality of types of light sources for emitting excitation light having different wavelengths are prepared so that the wavelength of the excitation light L can be appropriately selected according to the type of the phosphor that labels the non-test substance. It is configured so that it can be replaced. In addition, it is good also as a structure which arrange | positions LED which light-emits by multiple types of wavelength alternately in one light source, and light-emits only LED which emits the light of a desired wavelength suitably, or a white light source, such as a halogen lamp or white LED, A planar light source may be configured by combining a plurality of spectral filters, and the wavelength band of the excitation light L may be switched by appropriately replacing the spectral filters.

  The subject moving unit 60 moves the subject placement unit 40 in the direction of the arrow Z, and includes a drive unit 61 such as a motor, a shaft 62, a pulley 63, a belt 64, and the like. When the drive unit 61 is driven, the subject placement unit 40 moves in the arrow Z direction via the shaft 62, the pulley 63, and the belt 64. The operation of the subject moving unit 60 is controlled by a movement control unit 101 described later. The subject moving unit 60 includes, for example, an encoder, and detects subject position information PI from the amount of movement of the subject placement unit 40 and outputs it to the movement control means 101. That is, the drive unit 61 of the subject moving unit 60 also functions as a subject position detection unit. Note that it is preferable that the drive unit 61 is made of a pulse motor because control can be easily performed.

  In addition, when changing the photographing distance (conjugate length) SD between the subject 5 and the imaging surface 31a, a method of moving the imaging unit 31 and a method of moving the subject 5 side are conceivable, but the imaging unit 31 including a cooling unit is conceivable. Since the mechanism is complicated to move, it is preferable that the subject placement unit 40 is moved as in the present embodiment.

  FIG. 3 is a block diagram showing a preferred embodiment of the photographing control apparatus of the present invention. The photographing control apparatus 100 will be described with reference to FIGS. Note that the configuration of the imaging control apparatus 100 as shown in FIG. 3 is realized by executing an imaging control program read into the auxiliary storage device on a computer (for example, a personal computer).

  The imaging control apparatus 100 controls the imaging apparatus 10. The imaging control apparatus 100 receives the information regarding the fluorescence wavelength of the phosphor to be imaged and the imaging conditions input from the input unit 2, and the input receiving unit 101 receives the information. A photographing control unit 102 for controlling the photographing unit 30 and the light source 50 based on the photographing conditions, an image obtaining unit 103 for obtaining image information photographed by the photographing device 10, and a fluorescent image display unit based on the image information. 3, a display control unit 104 that displays the object, a movement control unit 105 that controls the subject moving unit 60, and a table T that stores the relationship between the type of phosphor and the optimum photographing distance for the wavelength of the phosphor. A storage unit 106 and a chromatic aberration correction instruction unit 107 that instructs the movement control unit 105 to perform fine adjustment for chromatic aberration correction are provided.

  When the input control unit 102 receives an instruction from the input unit 2 to shoot a fluorescent image of a subject in which a plurality of analyte substances labeled with a plurality of different phosphors are distributed, the imaging control unit 102 The photographing unit 30 and the light source 50 are controlled so as to sequentially photograph fluorescent images of different wavelengths with respect to the subject.

  The image acquisition unit 103 sequentially captures image information of images captured by the imaging device 10 into the imaging control device 100, and the display control unit 104 displays a fluorescent image on the display unit based on the acquired image information. When a plurality of fluorescent images having different wavelengths are photographed for one subject, an image obtained by superimposing the plurality of fluorescent images is displayed on the display unit.

  The movement control unit 105 has a function of controlling the operation of the subject moving unit 60 and obtaining the position information PI of the subject 5 from the operation amount of the driving unit 61 of the subject moving unit 60. Specifically, the movement control unit 105 has position information of the imaging unit 30 fixed to the housing 20 in advance, and is based on the position information of the imaging unit 30 and the position information PI of the subject placement unit 40. The shooting distance SD is obtained.

  The movement control unit 105 has a function of controlling the subject moving unit 60 so as to change the shooting distance SD in accordance with an instruction input from the input unit 2 by the user, and also in accordance with an instruction from the chromatic aberration correction instruction unit 107. It has a function of controlling the subject moving unit 60 to finely adjust the SD. Accordingly, the user can move the subject holding position up and down according to the subject for each subject having different dimensions, and can capture an image with a suitable size even for subjects of different dimensions.

  The chromatic aberration correction instructing means 107, when the user inputs an instruction for sequentially photographing the phosphor image of the first wavelength and the phosphor image of the second wavelength as the photographing of the subject 5, is input from the input means 2. The information about the type of the phosphor is received from the input receiving unit 101 that has received the type of the phosphor, and the fine adjustment amount is input to the movement control unit 105 with reference to the table T of the storage unit 106. In response to the fine adjustment instruction by the chromatic aberration correction instruction unit 107, the movement control unit 105 causes the subject moving unit 60 to capture the phosphor image of the first wavelength and before capturing the phosphor image of the second wavelength. The subject placement unit 40 is moved in the optical axis direction (arrow Z direction) of the imaging lens 32 by a fine adjustment amount to finely adjust the shooting distance SD. That is, in the photographing system of the present embodiment, the chromatic aberration correction unit is configured by the chromatic aberration correction instruction unit 107, the movement control unit 105, and the subject moving unit 60.

FIG. 4 shows an example of the table T stored in the storage unit 106. Table T correlates a combination of phosphors that generate fluorescence of different wavelengths, which are used simultaneously as labels on the same subject, and an optimum photographing distance difference (fine adjustment amount) caused by the wavelength difference between the two fluorescences. Data is stored. The table T shown in the figure shows that the fine adjustment amount in the case of the combination of the phosphor A and the phosphor B is d _AB , and the fine adjustment amount in the combination of the phosphor A and the phosphor C is d _AC. Holding.

Specifically, a fluorescent image AD by fluorescence from the phosphor A and a fluorescence image BD by fluorescence from the phosphor B are sequentially photographed with respect to a subject in which substances labeled with the phosphor A and the phosphor B are distributed. When instructed, the chromatic aberration correction instructing means 107 refers to the first column of the table T, and after taking the fluorescent image AD and before photographing the phosphor B, the optical axis of the lens is adjusted by the fine adjustment amount d _AB. The movement control means 105 is instructed to move the subject in the direction (the arrow Z direction in the figure). The movement control means 105 controls the subject moving unit 60 according to the instruction to move the subject 5 by the fine adjustment amount d _AB . Thereby, fluorescent images AD and BD in which the chromatic aberration of magnification due to the wavelength difference between both phosphors A and B is corrected can be obtained. For example, as shown in FIG. 5, the display control unit 104 displays the sequentially captured fluorescent image AD and fluorescent image BD on the display unit 3 and displays the image ABD on which both images are superimposed on the display unit 3. It has a function to display.

  The combination of phosphors and the fine adjustment amount held by the table T can be obtained based on the lens design information of the lens unit 32 that is an imaging lens. From the lens design information, the chromatic aberration of magnification caused by the wavelength difference of the fluorescence imaged through the lens, that is, how much the shooting distance should be changed to obtain the image of the same magnification for the fluorescence of different wavelengths Therefore, a table of phosphor combinations and fine adjustment amounts can be created.

  In addition, when the lens design information is unknown, before taking a fluorescent image, the shooting distance at which the same magnification image can be obtained for a plurality of fluorescence used as a label is measured, and the shooting distance between each fluorescence is measured. A table may be created by obtaining the difference (conjugate length difference).

  Note that lateral chromatic aberration also occurs due to differences in the thickness and refractive index of the excitation light cut filter 35. Since which excitation light cut filter is used in combination with each phosphor is usually uniquely determined, the amount of fine chromatic aberration generated by the excitation light cut filter is incorporated in advance in the fine adjustment amount held by the table T. Just go.

Next, the operation of the imaging system 1 of this embodiment will be described. First, the user opens the lid 22 of the housing 20 in FIG. 1, and the subject 5 in which a plurality of substances to be inspected labeled with two or more different types of phosphors are distributed is placed on the placement surface 42 of the subject placement unit 40. Set to a predetermined position. Here, it is assumed that the phosphor A and the phosphor B are used as labels, and the fluorescent image AD by the phosphor A and the fluorescent image BD by the phosphor B are sequentially photographed from the subject 5. Therefore, first, as incident light source 52, and it sets a light source that emits excitation light L _A optimum wavelength to excite the phosphors A. Thereafter, the user closes the lid 22 to prevent the outside light from entering the housing 20.

  In addition, the user inputs photographing conditions including sequentially photographing fluorescent images of the phosphor A and the phosphor B with the input unit 2 to the photographing control device 100.

  The input receiving unit 101 of the imaging control apparatus 100 transmits an instruction to start imaging to the imaging control unit 102 and the movement control unit 105, and imaging is started.

  Prior to photographing the phosphor A, the user operates the subject moving unit 60 on the photographing control apparatus 100 side to operate the subject moving unit 60 to determine the photographing distance SD. At this time, the shooting distance SD is determined by operating the user so as to obtain a shot image of a desired size while viewing the preview screen of the subject 5 displayed on the display unit 3. When the subject 5 moves, focus servo by the lens unit 32 is appropriately performed. When the subject moving unit 60 operates, position information PI is sent from the subject moving unit 60 to the movement control means 101.

When the subject 5 is positioned, imaging conditions such as the exposure time are set on the computer side. Thereafter, the imaging unit 30 captures the fluorescent image AD with the phosphor A under the imaging conditions set by the user to capture the subject 5. Excitation light L _A light source 52 is driven is irradiated to the object 5, the inspection substance labeled with a fluorescent substance A fluoresces when excited, while not fluoresce those labeled with a fluorescent substance B . Fluorescence emitted from the phosphor A is incident on the imaging surface 31a of the imaging unit 31 by the lens unit 32, and is detected photoelectrically by each imaging element of the imaging unit 31. Incidentally, the imaging unit 31 for is provided with an excitation light cut filter 35, the excitation light L _A reflected by the object 5 is cut by the excitation light cut filter 35 is provided between the object 5 and the imaging unit 31 Not incident.

  The two-dimensional image information (fluorescence image) AD captured by the imaging unit 31 is output to the display unit 3 by the display control unit 104 via the image acquisition unit 103 of the imaging control device 100. At this time, only the image a corresponding to the substance emitting fluorescence A among all the inspected substances on the subject 5 is displayed brightly on the image.

After the shooting of the phosphor A, the user opens the lid 22 of the housing 20, a light source 52, replace the light source for emitting excitation light L _B of optimal wavelength to excite the phosphors B, recapped 22 Close.

Next, before photographing the phosphor B, the chromatic aberration correction instructing unit 107 refers to the table T stored in the storage unit 106 based on the information regarding the phosphor previously input, and finely adjusts the photographing distance SD. The amount is read and the movement control means 105 is instructed to change the shooting distance by d _AB . Based on this instruction, the movement control means 105 causes the subject moving unit 60 to move the position of the subject 5 in the arrow Z direction by the fine adjustment amount d _AB .

Thereafter, photographing of the phosphor B is performed in the same manner as in the case of the phosphor A. Excitation light L _B light source 52 is driven is irradiated to the object 5, the inspection substance labeled with a fluorescent substance B fluoresces when excited, while not fluoresce those labeled with a fluorescent substance A . The fluorescence emitted from the phosphor B is incident on the imaging surface 31a of the imaging unit 31 by the lens unit 32 and is detected photoelectrically by each imaging element of the imaging unit 31. Incidentally, the excitation light cut filter 35 provided between the subject and the imaging unit is appropriately changed according to the wavelength of the excitation light, here, there is a cut filter for excitation light L _B.

  Two-dimensional image information (fluorescence image) BD imaged by the imaging unit 31 is output to the display unit 3 by the display control unit 104 via the image acquisition unit 103 of the imaging control device 100. At this time, only the image b corresponding to the substance that emits fluorescence B among all the substances to be inspected on the subject 5 is displayed brightly on the image.

  Thereafter, the display control means 104 displays an image ABD in which the fluorescent image AD and the fluorescent image BD are superimposed on the display unit 3.

  The imaging system 1 has a table T that holds a combination of phosphors and a fine adjustment amount at that time, a magnification after capturing a fluorescent image of the first wavelength based on the table T, and before capturing a fluorescent image of the second wavelength. Since the chromatic aberration correction instructing means 107, the movement control means 105, and the subject moving section 60 that constitute the chromatic aberration correcting means for finely adjusting the position of the subject so as to correct the chromatic aberration are provided, the area between the fluorescent image AD and the fluorescent image BD is provided. However, there is no difference in image size due to the lateral chromatic aberration of the lens. Therefore, an accurate distribution state of the labeled analyte substance on the subject 5 can be obtained in the image ABD obtained by superimposing the images AD and BD.

As a specific example, a case where phosphor A is Cy3 and phosphor B is Cy5 will be described. In this case, the excitation light _{L A} for exciting the Cy3, with those having a center wavelength of approximately 532 nm, the excitation light _{L B} to excite the Cy5, used having a center wavelength of approximately 635 nm. At this time, when the fluorescence near 575 nm of the fluorescence emitted by Cy3 and the fluorescence near 690 nm of the fluorescence emitted by Cy5 are photographed at the same photographing distance SD, respectively, an image of receiving and imaging the fluorescence of Cy3 at 575 nm, Due to the chromatic aberration characteristics of the lens part 32, the image of Cy5 fluorescence is larger than the image of Cy3 fluorescence due to the chromatic aberration characteristic of the Cy5 fluorescence received at 690nm. Therefore, after shooting Cy3 and before shooting Cy5, the shooting distance SD is finely adjusted to be slightly longer so that the Cy5 image has the same size as the Cy3 image.

Table 1 shows the photographing distance for each wavelength for each desired magnification when the lens unit used as the imaging lens of the present embodiment is used.

  A specific fine adjustment amount in the case where a lens having the characteristics shown in Table 1 using Cy3 and Cy5 as described above will be described. For example, in order to obtain an image with a magnification of −0.1551, the imaging distance when imaging at a Cy3 wavelength of 575 nm is 400.51 mm, and when the same magnification is obtained at a Cy5 wavelength of 690 nm, the imaging distance is 401. .78 mm. Therefore, at the time of shooting Cy5, an image having the same size can be obtained by making the shooting distance longer by 1.27 mm than the shooting distance of Cy3, that is, an image with corrected chromatic aberration of magnification can be obtained.

  In the present photographing system 1, it is preferable that the photographing condition of photographing once performed is stored in the storage unit 106 as a template. That is, when shooting is performed, shooting is performed by combining the input shooting conditions input by the user, the set shooting distance SD, the shooting position PI at that time, and the fine adjustment amount read from the table T. A condition is stored in the storage unit 106 as a template. As a result, when the user inputs the same input photographing condition from the input unit 2 next time, the photographing control unit 102, the movement control unit 105, and the chromatic aberration correction instruction unit 107 are stored in the storage unit 106 as indicated by the dotted line in FIG. The same shooting conditions can be reproduced by reading the conditions stored in the.

The perspective view which shows preferable embodiment of the imaging | photography system of this invention The top view which shows schematic structure of the imaging device of FIG. The block diagram which shows preferable embodiment of the imaging | photography control apparatus of the imaging | photography system of this invention. Data table on phosphor type and fine adjustment amount The schematic diagram which shows an example of the fluorescence image by 1st fluorescence, the fluorescence image by 2nd fluorescence, and the fluorescence image which superposed them

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Shooting system 2 Input means 3 Display part 5 Subject 10 Shooting device 30 Shooting unit 31 Imaging part (imaging element)
32 Lens (imaging lens)
35 Excitation light cut filter 50 Light source 60 Subject moving means 100 Imaging control device 101 Input receiving means 102 Imaging control means 103 Image acquisition means 104 Display control means 105 Movement control means 106 Storage means 107 Chromatic aberration correction instruction means T Table SD Imaging distance (conjugate) Long)

Claims (5)

An imaging apparatus including a housing that accommodates a subject in which a plurality of areas in which a substance labeled with a phosphor is distributed is set, a photographing unit that captures a fluorescent image of the subject, and controls the operation of the photographing unit In a shooting system equipped with a shooting control device,
The imaging unit includes an imaging element that acquires a fluorescent image of a subject, and an imaging lens that forms an image of the fluorescent image on an imaging surface of the imaging element;
The imaging control device controls the imaging unit to capture a fluorescent image of a second wavelength different from the first wavelength of the subject after capturing a fluorescent image of the first wavelength of the subject, Obtaining an image obtained by superimposing the fluorescence image of the first wavelength and the fluorescence image of the second wavelength;
After taking the fluorescence image of the first wavelength, before taking the fluorescence image of the second wavelength, the chromatic aberration of magnification of the imaging lens caused by the wavelength difference between the first wavelength and the second wavelength And a chromatic aberration correcting means for moving at least one of the image sensor and the subject in the optical axis direction of the imaging lens so as to correct the image. A storage unit storing a table holding a relationship between a chromatic aberration of magnification based on a difference between the first wavelength and the second wavelength and an optimum difference in photographing distance at each wavelength;
The photographing system according to claim 1, wherein the chromatic aberration correcting unit moves at least one of the image sensor and the subject based on the relationship.   The imaging system according to claim 2, wherein the relationship held in the table is obtained based on design information of the imaging lens.   3. The photographing system according to claim 2, wherein the relationship held in the table is obtained by actual measurement before photographing the fluorescent image.   5. The photographing system according to claim 1, wherein the chromatic aberration correcting means includes a pulse motor as means for moving at least one of the image sensor and the subject.

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