Abstract:
A diagnosis device includes: a pretreating portion configured to suspend tissue to obtain a suspension; and an analyzing portion configured to analyze the suspension.

Description:
BACKGROUND OF THE INVENTION 
       [0001]    The present invention relates to a diagnosis device and method which, when extracted tissue is loaded, can be automatically used in diagnosis such as diagnosis of cancer. 
         [0002]    After a pathological specimen is prepared, a cytotechnologist or a pathologist performs pathology by using a tissue slice. 
         [0003]    In the case where pathology is performed as described above, however, the cytotechnologist or the pathologist is required to have a skillful technique, and there is a problem in that the pathology is performed in a different manner depending on the difference in the skills. 
         [0004]    In processes from tissue extraction to pathology, techniques such as tissue fixation, preparation of section, and stain are necessary, and works must be continued to be manually performed for a certain time period. By contrast, a related-art device which isolates cells from a tissue slice to obtain isolated cells is known. Such a device is used for obtaining a cell suspension by a movable dissociated element such as a blade, and configured by a single container (see JP-A-2007-505631, particularly [0025]). 
         [0005]    Therefore, it is difficult to obtain suspension of a desired state by changing or adjusting steps such as tissue fixation, preparation of section, and stain. Furthermore, the suspension must be supplied to a flow cytometer while the container is held by a hand. Therefore, development of an automated pathology device is requested. 
       SUMMARY 
       [0006]    It is therefore an object of the invention to provide a diagnosis device and method which are configured so as to, when extracted tissue is loaded, automatically analyze suspended cells. 
         [0007]    In order to achieve the object, according to the invention, there is provided a diagnosis device comprising: 
         [0008]    a pretreating portion configured to suspend tissue to obtain a suspension; and 
         [0009]    an analyzing portion configured to analyze the suspension. 
         [0010]    The pretreating portion may include a cleansing portion configured to clean the tissue. 
         [0011]    The pretreating portion may include a chopping portion configured to chop the tissue. 
         [0012]    The pretreating portion may include a filtering portion which filters the suspension. 
         [0013]    The diagnosis device may further include an enzyme treating portion configured to inject an enzyme into the suspension, thereby performing an enzyme treatment. 
         [0014]    The diagnosis device may further include an enzyme treating portion configured to inject an enzyme into the suspension that is filtered by the filtering portion, thereby performing an enzyme treatment. 
         [0015]    The diagnosis device may further include a centrifugal separating portion configured to apply centrifugal separation to the suspension that is filtered by the filtering portion, thereby removing supernatant. 
         [0016]    The diagnosis device may further include a centrifugal separating portion configured to apply centrifugal separation to the suspension that is subjected to the enzyme treatment performed by the enzyme treating portion, thereby removing supernatant. 
         [0017]    The diagnosis device may further include a centrifugal separating portion configured to apply centrifugal separation to the suspension that is filtered by the filtering portion, thereby removing supernatant. 
         [0018]    The diagnosis device may further include a centrifugal separating portion configured to apply centrifugal separation to the suspension that is subjected to the enzyme treatment performed by the enzyme treating portion, thereby removing supernatant. 
         [0019]    The pretreating portion may include a staining portion configured to stain the suspension. 
         [0020]    The analyzing portion may be a flow cytometer. 
         [0021]    In order to achieve the object, according to the invention, there is also provided a diagnosis method comprising: 
         [0022]    suspending tissue to obtain a suspension; and 
         [0023]    analyzing the suspension. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0024]      FIG. 1  is a front view of a state where a flow cytometer an embodiment of a diagnosis device of the invention is exposed. 
           [0025]      FIG. 2  is a perspective view in which the interior of the embodiment is exposed. 
           [0026]      FIG. 3  is a perspective view showing a pretreating portion of the embodiment, in a state where the portion is exposed. 
           [0027]      FIG. 4  is a flowchart showing an example of the operation of the pretreating portion of the embodiment. 
       
    
    
     DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS 
       [0028]    Hereinafter, an embodiment of the diagnosis device of the invention will be described with reference to the accompanying drawings. In the figures, identical components are denoted by the same reference numerals, and duplicated description is omitted.  FIG. 1  is a front view of the diagnosis device,  FIG. 2  is a perspective view of the diagnosis device, and  FIG. 3  is a perspective view of a pretreating portion  10  which constitutes main portions of the diagnosis device. As shown in  FIGS. 1 and 2 , the diagnosis device includes a flow cytometer  60 , in addition to the pretreating portion  10 . 
         [0029]    In a front panel  11  of the pretreating portion  10 , a slide door  12  for loading extracted tissue is disposed, and operation buttons  13  through which instructions for pretreatment is to be given are provided. A base  14  is disposed in a lower portion of the rear face of the front panel  11 . Two rails  15  which extend from the rear side of the front panel  11  are disposed on the base  14 . 
         [0030]    A buffer tank  16  is disposed in a position between the two rails  15  so as to be slidable along the rails  15 . A driving source is disposed in at least one of the buffer tank  16  and the rails  15 . 
         [0031]    An upper portion of the buffer tank  16  is opened. Above a position (first position) where the buffer tank  16  is moved closest to the base  14 , for example, a basket  17  which is formed by a stainless steel mesh is hung by a pipe  18  in a manner that the basket is made vertically movable by the pipe  18  so that the basket can be fluctuated at a position of a predetermined height in the buffer tank  16 . A platform scale  19  is disposed in an upper portion of the pipe  18 , and the weight of tissue which is loaded into the basket  17  can be measured. The pipe  18  is connected through a pump  21  to a reservoir tank  29  for a buffer solution such as PBS (phosphoric acid buffered saline), and the buffer solution can be supplied from the reservoir tank  29  into the buffer tank  16 . 
         [0032]    A waste solution line  20  is coupled to a bottom portion of the buffer tank  16 , and communicates with a waste solution tank  22  through the pump  21 . A hole  23  is formed in a bottom portion of the basket  17 , and a slide cover  24  having a mechanism which makes the hole  23  openable and closable is disposed. The buffer tank  16 , the basket  17 , the pipe  18 , the reservoir tank  29  for the buffer solution, and the like constitute a cleansing portion which cleanses the tissue with the solution. 
         [0033]    A cutter  26  which has a blade  25  at the tip end is hung above a position (second position) where the buffer tank  16  is moved remotest on the rails  15  from the base  14 . The cutter  26  includes a movement mechanism which vertical moves. The blade  25  is coupled to a motor  28  via a shaft  27  so as to be rotated. The buffer tank  16  and the cutter  26  constitute a chopping/suspending portion which receives a tissue slice from the above-described cleansing portion, and which chops the tissue slice with the rotary blade to suspend the tissue. 
         [0034]    At a third position which is more separated from the base  14  than the second position, and which is at an approximately same level as the rails  15 , a nylon mesh filter  30  is hung by an arm  31 . An aspiration and discharge device  32  is disposed above the nylon mesh filter  30 . An aspiration line  33  which is coupled to the bottom portion of the buffer tank  16  is connected to the aspiration and discharge device  32  so that the suspension in the buffer tank  16  can be aspirated through the aspiration line  33 . 
         [0035]    A nozzle  34  is disposed in the lower end face of the aspiration and discharge device  32 . The suspension which is aspirated from the buffer tank  16  can be discharged through the nozzle  34  into the nylon mesh filter  30 . An infundibulum  35  is disposed below the nylon mesh filter  30 . A gutter  36  is disposed at the tip end of the infundibulum  35 . The nylon mesh filter  30  and the aspiration and discharge device  32  constitute a filtering portion which performs filtration by means of the mesh. The nylon mesh filter  30  can be moved to a position outside the space between the nozzle  34  and the infundibulum  35 , by a movement mechanism coupled to the arm  31 . 
         [0036]    A centrifugal separator  40  is placed on a floor face at a middle position between the third and second positions. The centrifugal separator  40  includes a rotation shaft  42  in the middle of a bottom portion of a bottomed cylindrical basket  41 , and a motor which rotates the rotation shaft  42 . Several solution reservoirs  43  are placed in the periphery of the rotation shaft  42 . Each of the solution reservoirs  43  can be positioned to a tip end portion of the gutter  36  by the rotation to receive the supply of the suspension, and is provided with a cover which is opened and closed by an opening and closing mechanism. The centrifugal separator  40  constitutes a centrifugal separating portion which applies centrifugal separation to the suspension, thereby removing supernatant, and which further performs a suspending process. 
         [0037]    In a space which is above the solution reservoirs  43 , and which is located at a position where the suspension is not supplied from the tip end portion of the gutter  36 , a staining portion  50  is disposed to inject an enzyme and a fluorescent stain solution into the suspension, thereby performing an enzyme/stain process. The staining portion  50  is configured so as to be vertically movable. An aspiration/discharge line  51  for the cell suspension is connected to the staining portion  50 . The staining portion  50  is configured so that, when the portion is downward moved, the tip end of the aspiration/discharge line  51  is lowered to a position where the line can aspirate the cell suspension in the solution reservoirs  43  which is below the staining portion  50 . 
         [0038]    An enzyme/stain solution supply line  52  is connected to the staining portion  50 , and communicates through the pump  21  with an enzyme/stain solution tank  53 . The staining portion  50  aspirates a stain solution from the enzyme/stain solution tank  53  through the enzyme/stain solution supply line  52  and the pump  21 , and mixes the stain solution with the cell suspension which is aspirated from the aspiration/discharge line  51  to perform an enzyme treatment. The cell suspension which has undergone the enzyme treatment is supplied to a flow cell  61  of the flow cytometer  60  through a cell suspension supply line  54 . 
         [0039]    In the above-described pretreating portion  10 , a pretreatment controlling portion  90  controls the various portions. The pretreatment controlling portion  90  is configured by a computer, and controls the various portions of the pretreating portion  10 . In the embodiment, for example, the control is performed by a program corresponding to the flowchart shown in  FIG. 4 . Therefore, the operation will be described with reference to the flowchart. 
         [0040]    First, the examiner opens the slide door  12 , loads extracted tissue into the basket  17 , closes the door  12 , and operates a start button of the operation buttons  13 . As a result, a pretreating step S 11  is performed. In the pretreating step S 11 , the tissue is weighed by the platform scale  19 , and the pretreatment controlling portion  90  receives and holds the weighing result. 
         [0041]    In the pretreating step S 11 , next, the buffer solution is supplied from the reservoir tank  29  for the buffer solution into the buffer tank  16  through the pipe  18 , and the basket  17  is lowered by the pipe  18  to be immersed in the buffer solution, and fluctuated so that the tissue is cleansed for a predetermined time period. Then, the basket  17  is raised by the pipe  18 , and the cleansing solution (buffer solution) is recovered by the pump  21  from the waste solution line  20  to the waste solution tank  22 . 
         [0042]    In the pretreating step S 11 , furthermore, the buffer solution the amount of which corresponds to the weight of the tissue is supplied through the pipe  18  into the buffer tank  16 , the basket  17  is lowered by the pipe  18  to be immersed in the buffer solution, and the slide cover  24  is opened, so that the tissue slice is loaded into the buffer solution of the buffer tank  16 . 
         [0043]    Next, a chopping/suspending step S 12  for the tissue slice is executed. In the chopping/suspending step S 12 , the buffer tank  16  is moved along the rails  15  to be located at a position immediately below the blade  25  of the cutter  26 . Then, the cutter  26  is lowered into the buffer solution of the buffer tank  16 , and the blade  25  is rotated to chop and suspend the tissue. The pretreatment controlling portion  90  controls the speed of rotation and the time period in accordance with previously given information indicative of the kind of the tissue (for example, a plant, an animal, or a human, or a region (an internal organ, or skin)), and the weight of the tissue which is measured as described above, thereby obtaining a suspension of a desired state. 
         [0044]    Next, the process proceeds to a filtering step S 13 . Before the filtering step S 13 , the cutter  26  is raised and moved to the home position. In the filtering step S 13 , the nylon mesh filter  30  is positioned between the nozzle  34  of the aspiration and discharge device  32  and the infundibulum  35 , and the aspiration and discharge device  32  aspirates the solution of the buffer tank  16  through the aspiration line  33 , and discharges the suspension from the nozzle  34  toward the nylon mesh filter  30  to perform filtration. The solution reservoirs  43  of the centrifugal separator  40  are sequentially positioned in the tip end portion of the gutter  36 , and the filtered suspension is supplied through the gutter  36  to the solution reservoirs  43  to be stored therein. 
         [0045]    Next, a staining step S 14  is executed. In the staining step S 14 , the staining portion  50  is lowered, and sequentially aspirates the suspension stored in tho solution reservoirs  43  of the centrifugal separator  40  by means of the aspiration/discharge line  51  to hold them. An incubation (staining process) in which a predetermined amount of the stain solution is supplied from the enzyme/stain solution tank  53  through the enzyme/stain solution supply line  52  and the pump  21  to be mixed with the suspension is carried out. The pretreatment controlling portion  90  controls the supply amounts of the enzyme and the stain solution so as to attain an adequate dilution degree corresponding to the previously given information indicative of the kind of the tissue, the weight of the tissue which is measured in the above, etc. 
         [0046]    Next, a centrifugal separating step S 15  is performed. In the centrifugal separating step S 15 , the staining portion  50  discharges the cell suspension which has undergone the staining process, into the solution reservoirs  43  of the centrifugal separator  40  through the aspiration/discharge line  51 , and thereafter is raised to return to the home position. The covers of the solution reservoirs  43  are closed, and centrifugal separation is performed by rotation about the rotation shaft  42 . The pretreatment controlling portion  90  controls the speed of rotation and the time period in accordance with the previously given information indicative of the kind of the tissue, the weight of the tissue which is measured in the above, etc. Then, the staining portion  50  is again lowered, the staining portion  50  sequentially aspirates the cell suspension in which supernatant is removed from the suspension stored in the solution reservoirs  43  of the centrifugal separator  40 , by means of the aspiration/discharge line  51 , and supplies the cell suspension to the flow cell  61  of the flow cytometer  60 . 
         [0047]    The process by the flowchart of  FIG. 4  is a mere example. At least one of the filtering step S 13 , the staining step S 14 , and the centrifugal separating step S 15  may be omitted. When the filtering step S 13  is to be omitted, the nylon mesh filter  30  may be placed at a retracted position. When the staining step S 14  is to be omitted, the enzyme may not be supplied. When the centrifugal separating step S 15  is to be omitted, the centrifugal separating step S 15  may not be performed. 
         [0048]    The sequence of the staining step S 14  and the centrifugal separating step S 15  may be changed. When a suspension discharge line extending from the staining portion  50  into the buffer tank  16  is disposed, the filtering step S 13  may be repeated many times. 
         [0049]    The progress of the steps may be set as a program in the pretreatment controlling portion  90 . Alternatively, each time one of the steps is ended, the examiner may instruct the next step by using the operation buttons  13 . In the alternative, the processes from the loading of tissue to the supply of the cell suspension to the flow cytometer  60  are not fully automatically performed, but, in the steps, the process contents such as the rotation number, and the process time period are automatically determined, and a cell suspension of the desired state can be obtained and supplied to the flow cytometer. 
         [0050]    The flow cytometer  60  includes the flow cell  61 , laser irradiation devices  62  to  64 , dichroic mirrors  65  to  67 , a forward-scattered light detection sensor  68 , a mirror block  70 , a side-scattered light detection sensor  71 , and fluorescence detection sensors  72  to  75 . In the flow cell  61 , the cell suspension flows through the middle of a sheath liquid flow, and cells flow through a laser irradiation portion  69  while being arranged in one row, and then discharged from the flow cell  61  to be formed as a liquid droplet, with the result that the cells are sorted. 
         [0051]    Laser beams emitted from the laser irradiation devices  62  to  64  reach the laser irradiation portion  69  of the flow cell  61  through the respective dichroic mirrors  65  to  67 . The laser beams are scattered by the cells flowing through the laser irradiation portion  69 . 
         [0052]    The forward-scattered light detection sensor  68  is placed at a position which is on an extension of the optical path of the laser beams reaching the laser irradiation portion  69  through the dichroic mirrors  65  to  67 . The mirror block  70  is placed in an optical path of side-scattered light which intersects at 90 degrees with the above-described optical path. 
         [0053]    The mirror block  70  separates the light beams in accordance with the wavelengths to be detected by the side-scattered light detection sensor  71  and the fluorescence detection sensors  72  to  75 , and guides the separated light beams to the side-scattered light detection sensor  71  and the fluorescence detection sensors  72  to  75 . A cell scatters forward-scattered light of an intensity corresponding to the size of the cell. The forward-scattered light detection sensor  68  which receives the forward-scattered light detects a potential corresponding to the intensity, and sends it to a computer  80 . The side-scattered light detection sensor  71  and the fluorescence detection sensors  72  to  75  introduce light of the respective wavelengths to be detected, convert the light to potentials, and send them to the computer  80 . 
         [0054]    In the computer  80 , the potentials sent from the forward-scattered light detection sensor  68 , the side-scattered light detection sensor  71 , and the fluorescence detection sensors  72  to  75  are A/D converted to form histograms, and a dot plot in which the forward-scattered light histogram and the side-scattered light histogram are overlapped with each other is produced and displayed on a displaying portion  81 . In the computer  80 , furthermore, fluorescence data are produced on the basis of the outputs of the fluorescence detection sensors  72  to  75 , and displayed on the displaying portion  81 . 
         [0055]    Each of the cells is classified into a normal cell and a cancer cell depending on the measurements of forward-scattered light, side-scattered light, and various fluorescences, and a cancer diagnosis of the tissue can be performed on the basis of the cells in the suspension. 
         [0056]    Of course, the flow cytometer  60  may further have other related-art functions other than the above-described functions, and output various analysis data required in various diagnoses. As described above, according to the invention, when an extracted tissue slice is loaded, isolated and suspended cells can be automatically supplied to the flow cytometer, and a histogram and a dot plot of the cell sizes and fluorescences can be produced and output. Therefore, data necessary for various diagnoses can be obtained without requiring any skill. 
         [0057]    According to an aspect of the invention, a diagnosis device and system can automatically analyze suspended cells when tissue is loaded therein.