Patent Publication Number: US-2018044625-A1

Title: Incubator and incubator system with clean bench function

Description:
TECHNICAL FIELD 
     The present invention relates to an incubator having both a function of a clean bench used for working in an open space system and a function of the incubator used for culturing in a closed space system. 
     BACKGROUND ART 
     Generally, in culturing cells or tissues, a clean bench and incubator are used, where the clean bench provides a sterile space for an operator to manipulate cells etc. and the incubator provides a space to culture cells etc. after the operation in clean bench. The clean bench feeds sterile gas into a working space and thereby produces a sterile working space. The operator inserts hands into the working space in clean bench through an opening site to manipulate cells etc. that placed on the working space. The incubator is equipped with a sealed up the inside space, and is capable of controlling and regulating the gas composition and temperature inside incubator as desired according to experimental conditions for the culture of cells etc. placed in the incubator (Patent Literature 1). In those experiments system, the operator manipulates cells etc. using the clean bench, and then needs to culture for the cells etc. by moving them to the incubator. 
     However, the clean bench and incubator are separate pieces of equipment, making it necessary for the operator to take a culture vessel out of the clean bench after working on the clean bench and move the culture vessel into the incubator. Also, depending on details of the experiment, cells etc. need to be kept immersed in the culture solution having, for example, a predetermined pH level, but the pH level may change with time and cease to satisfy a predetermined culture condition. Therefore, it is necessary to constantly monitor the pH level of the culture solution and to exchange the culture solution when the measurement of pH level is departed from a predetermined value. It is so troublesome to constantly monitor and exchange the culture condition, especially, the case of that it is not desirable to exchange the culture solution by opening and closing the sealed incubator kept at a predetermined gas composition ratio because the cells etc. might not be able to be cultured appropriately. 
     CITATION LIST 
     Patent Literature 
     Patent Literature 1: Japanese Patent Laid-Open No. 2012-90612 
     SUMMARY OF INVENTION 
     Technical Problem 
     The present invention has been made in view of the above problems and has an object to provide an incubator and an incubator system equipped with a clean bench function, allowing cells etc. to be manipulated and cultured. 
     Solution to Problem 
     An incubator with a clean bench function (the first unit e.g. in this system) according to a first aspect of the present application comprises: a chamber configured to be closable and openable; a gas supply unit adapted to supply a plurality of types of gas to the chamber; a concentration control unit adapted to control concentrations of the plurality of types of gas in the chamber; and a temperature control unit adapted to control temperature in the chamber. Preferably, the chamber has an opening, and includes an inner door adapted to close the entire opening of overall, and further includes an operation cover equipped in the inner door. Preferably, the inner door further includes an operation cover fixing hole used to attach the operation cover. Preferably, the operation cover is attached to the operation cover fixing hole using a circular fixture; and the circular fixture includes two half rings pivotally connected to each other at one end and linked together by a tension spring at the other end. The operation cover has been or can be sterilized, and is an arm cover and/or glove such as a glove with intact tips, a tubular glove with the tip cut off, or an arm cover made of non-cloth paper, cloth, silicone, etc. Preferably, the chamber has an opening and includes a sealing door to close the entire opening and shut off an inside the chamber from outside air. Besides, preferably the incubator further comprises a line (in/out) port used to pull tubes into the chamber from outside the chamber. Furthermore, preferably the chamber includes: an opening, an inner door adapted to close the entire opening, a sealing door adapted to close the entire opening and shut off an inside the chamber from outside air, and an elastic material equipped between the inner door and the opening and between the sealing door and the opening; the elastic material includes a channel extending in a width direction of the elastic material; and the channel makes up a line (in/out) port used to pull tubes into the chamber from outside the chamber. 
     The chamber may include: an opening, an inner door adapted to close the entire opening, a sealing door adapted to close the entire opening and shut off an interior of the chamber from outside air, a first elastic material attached to the chamber, which is positioned between the inner door and the opening, a second elastic material attached to the sealing door, which is positioned between the sealing door and the opening, and a third elastic material attached to the chamber, which is positioned between the sealing door and the opening; first elastic material, second elastic material, and third elastic material may include first channels, second channels, and third channels, respectively, extending in width directions of the respective elastic materials; and first channels, second channels, and third channels may make up a line (in/out) port used to pull tubes into the chamber from outside the chamber. 
     Positions of first channel, second channel, and third channel may be determined such that a straight line extending along first channel, second channel, or third channel does not cross an end of another adjacent channel. The plurality of types of gas may include oxygen (O 2 ) gas. Also, the plurality kinds of gas may include carbon dioxide (CO 2 ) gas; and the gas supply unit may be equipped with a CO 2  cylinder. The plurality of types of gas may include nitrogen (N 2 ) gas; and the gas supply unit may be equipped with an device adapted to generate N 2  or with a N 2  cylinder. Furthermore, the plurality of types of gas may include O 2  gas and CO 2  gas; and the concentration control unit may control O 2  concentration to be 25% or less and control CO 2  concentration to be 20% or less. Besides, the plurality of types of gas may include N 2  gas; and when the incubator with a clean bench function (the first unit e.g.) is used as a clean bench, the gas supply unit may supply N 2  gas into the chamber such that positive pressure is maintained in the chamber relative to outside pressure, but when the first unit is used as an incubator, the gas supply unit does not have to supply N 2  gas into the chamber. 
     The temperature control unit may control in the chamber to be in the wildly range from 4 degrees Celsius in air temperature to 50 degrees Celsius adding in room temperature. The chamber may have an opening, and further include an inner door adapted to close the entire opening, an operation cover attached in the inner door, and hooks installed in the chamber and adapted to be able to catch the operation cover. The hook may be a protruding rod-shaped material inside the chamber of lateral surface curved to the back surface. The incubator may further comprise a ultraviolet (UV) lamp installed inside the chamber. 
     According to second invention of the present application, the incubator system with the clean bench function (the first and second units in this system) applies to be possible to operate under the condition of very low temperature (from 4 degree Celsius in air temperature) too, and applies to be possible to regulate under the different conditions of both the gas composition and temperature including 4 degree Celsius at the same time too, whereby the functions of the second unit adds to the first unit that. The second unit combined with the first unit are comprised the incubator with the clean bench function (the second unit and/or the first unit); a low temperature chamber (the second unit) adapted to keep on the inside its temperature lower than that inside the first unit; and the work chambers installed in the low temperature chamber (the second unit); and the gas control units regulated gas concentration in the work chambers (the second unit). In particular, the combined with first and second invention is possible of exchange and regulate with gas each of different temperature in this system, that is the gas control unit is possible to regulate a constituent of gas in the work chamber inside the low temperature chamber in the second unit to exchange and match a constituent of gas in the first unit. The gas control unit may control concentrations of a plurality of types of gas in the work chamber (the second unit e.g.). Also, the gas control unit may regulate a constituent of gas in the work chamber inside the low temperature chamber in the second unit to be different from a constituent of gas in the first unit. Specifically in addition, the second invention in possible of exchange and regulate with gas both of each different temperature and constituent in this system. 
     Advantageous Effects of Invention 
     The present invention provides an incubator and an incubator system with a clean bench function (the first and second units in this system), allowing cells etc. to be manipulated and cultured. 
    
    
     
       BRIEF DESCRIPTION OF DRAWINGS 
         FIG. 1  is a diagram schematically showing an incubator with the clean bench function (the first unit e.g.) according to a first embodiment. It is illustrate here only with the first unit, however the second unit has similar applied functions of the first unit and in addition to other functions. 
         FIG. 2  is a block diagram of the incubator with the clean bench function (the first unit e.g.). 
         FIG. 3  is a perspective view of first elastic material and third elastic material. 
         FIG. 4  is a front view of second elastic material. 
         FIG. 5  is a front view of first elastic material, second elastic material, and third elastic material with tubes passed there through. 
         FIG. 6  is a perspective view of an inner door with holes attached the membrane with slits and the operation cover to insert into the gloves for operation, and a hook, shelves, etc. inside the chamber. 
         FIG. 7  is a perspective view of a hook having another shape. 
         FIG. 8  is a diagram schematically showing an incubator system (the first and/or second units in this system) according to a second embodiment. 
         FIG. 9  is a perspective view of fourth elastic material and fifth elastic material. 
         FIG. 10  is a block diagram of the incubator system (the first and/or second units in this system). 
         FIG. 11  is a front view of a circular fixture. 
         FIG. 12A  is a diagram showing a technique for attaching an operation cover to the circular fixture. 
         FIG. 13  is a diagram of the operation cover as viewed from outside the inner door. 
     
    
    
     REFERENCE SIGNS LIST 
     
         
         
           
               100  Incubator with clean bench function 
               110  Chamber 
               111  Opening 
               114  First hook 
               116  UV lamp 
               117  Outer sealing door 
               118  Inner door 
               119  Operation cover fixing hole 
               120  Third elastic material 
               121  Second elastic material 
               122  First elastic material 
               136  Operation cover 
               140  Gas concentration control unit 
               160  Temperature control unit 
               170  CO 2  cylinder 
               180  N 2  cylinder 
               190  N 2  gas generator 
               300  Circular fixture 
           
         
       
    
     DESCRIPTION OF EMBODIMENTS 
     First, an incubator  100  with a clean bench function (the first unit e.g.) according to an embodiment of the present invention will be described with reference to  FIGS. 1 to 7 . 
       FIG. 1  is a schematic diagram of the incubator  100  with a clean bench function (the first unit e.g.). The incubator  100  with a clean bench function mainly includes a chamber  110 , the gas concentration control unit  140 , a temperature control unit  160 , a CO 2  cylinder  170 , a N 2  cylinder  180 , and a N 2  gas generator  190 . The CO 2  cylinder  170 , N 2  cylinder  180 , and/or N 2  gas generator  190  makes up a gas supply unit. Note that for the sake of explanation, side faces of the chamber  110  are illustrated in such a way as to allow an inside the chamber  110  to be seen through. 
     The chamber  110  has the shape of a rectangular parallelepiped and includes an opening  111  formed by opening in a front of the chamber  110 , an outer sealing door (sealing door)  117  and an inner door  118  attached to the opening  111 , and a sensor  161 . The opening  111  is a part of the front in the chamber  110 , and includes a peripheral portion  112  on placement around the opening site. A first hook  114  and a sensor  161  are attached to each of two inner side surfaces  113  of the chamber  110 . Note that the first hooks  114  on the other side inner surfaces  113  is omitted in  FIG. 1  for simplification of explanation. A detailed shape of the first hooks  114  will be described later. A UV lamp  116  adapted to emit UV rays whose wavelength is suitable for sterilization is attached on upper site of an inner back surface  115  in the chamber  110 . Plural shelf boards  124  can be attached inside the chamber  110 . Depth of two shelf boards  124  close to the UV lamp  116  is approximately half-long depth of the another shelf boards inside the chamber  110  and it is a reason for adjusted such that the UV rays emitted by the UV lamp  116  will easily reach any desired place inside the chamber  110 . Consequently, all of the desired any place inside the chamber  110  is sterilized appropriately. The culture vessels  125  of a closed perfusion type and/or non-perfusion type etc. can be placed on the shelf boards  124 . Note that a non-illustrated heat insulating material is provided on a ceiling surface, the inner back surface  115 , the inner side surface  113 , and a bottom surface of the chamber  110 . 
     The sensor  161  includes a temperature sensor, CO 2  sensor, and O 2  sensor, and measures temperature, CO 2  concentration, and O 2  concentration in the chamber  110 . 
     The sealing door  117  and inner door  118  are attached to the opening  111  using pivotable hinges. The inner door  118  is made of a flat transparent polycarbonate plate length enough to completely close the opening  111  and provided with two arm cover/glove fixing holes (operation cover fixing holes)  119  perforating the polycarbonate plate in a thickness direction below a vertical center. The arm cover and/or glove are referred to as an operation cover. The two operation cover fixing holes  119  each has a circular shape with the same diameter, but unnecessary of the same it. The diameter is large enough to allow human arms to pass easily, reach anywhere inside the chamber  110  easily, and work easily. First elastic material  122  (see  FIG. 3 ) is attached on the circumference inside edge in the chamber  110  at the peripheral site  112  in the chamber  110  inner door  118 , where which on the most inside edge in the chamber  110  of the distance from outside edge, where is the distance from outside edge in its, and placed on the making overlapped area at the inner double doors by the closed the inner door  118  (see  FIG. 3, 5 ). Next, second elastic material  121  (see  FIG. 4 ) is attached on the circumference inside near the edge in the sealing door  117 , where the located of midsection in the peripheral site  112  is made appearance of overlapped area in nearly alternated site at the few distance of outside from first elastic material  122  and inside from third elastic material  120  by closed both the sealing door  117  and the inner door  118  (see  FIG. 5 ). And moreover, third elastic material  120  (see  FIG. 3 ) is attached on the outside edge of peripheral site  112  in the chamber  110 , and placed on the more slightly outside second elastic material  121  above-mentioned in the overlapped area (see  FIG. 5 ), The sealing door  117  is a substantially flat-plate metal door adapted to completely close the opening  111 , configured to be larger in area than the inner door  118  when viewed from the front. Details of the inner door  118 , sealing door  117 , third elastic material  120 , second elastic material  121 , and first elastic material  122  will be described later. 
     The gas concentration control unit  140  is connected with the chamber  110 , CO 2  cylinder  170 , N 2  cylinder  180 , and N 2  gas generator  190  through tubes. The gas concentration control unit  140  adjusts CO 2  gas supplied from CO 2  cylinder  170  and N 2  gas supplied from N 2  cylinder  180  and N 2  gas generator  190  to respective predetermined concentrations, and their mixes and supplies the gas mixture into the chamber  110  (see  FIG. 1, 2 ). The concentrations of CO 2  gas and N 2  gas are adjusted such that concentrations of CO 2 , N 2 , and O 2  in the chamber  110  will take predetermined values. Details of the concentration control unit  140  will be described later. 
     The temperature control unit  160  (see  FIG. 1 ), includes a heater and a refrigerating machine and maintains predetermined temperatures inside the chamber  110 , e.g., temperatures in the range from 4 degrees Celsius in air temperature to 50 degrees Celsius adding in room temperature, based on the temperature detected by the sensor  161  attached inside the chamber  110 . 
     CO 2  cylinder  170 , which is an airtight container, stores CO 2  gas. The tube is attached to CO 2  cylinder  170  via regulator  171 . The regulator  171  regulates a flow rate of CO 2  gas flowing out of CO 2  cylinder  170 . The tube connects CO 2  cylinder  170  to the gas concentration control unit  140 . CO 2  gas is sent from CO 2  cylinder  170  to the concentration control unit  140  through the tube. N 2  cylinder  180 , which is an airtight container, stores N 2  gas. The tube is attached to N 2  cylinder  180  via regulator  181 . The regulator  181  regulates flow rate of N 2  gas flowing out of N 2  cylinder  180 . The tube connects N 2  cylinder  180  to the gas concentration control unit  140 . N 2  gas is sent from N 2  cylinder  180  to the gas concentration control unit  140  through the tube. 
     N 2  gas generator  190  generates N 2  gas by separating N 2  from the atmosphere. The tube connects N 2  gas generator  190  to the concentration control unit  140 . N 2  gas is sent from N 2  gas generator  190  to the gas concentration control unit  140  through the tube. 
     The culture solution is supplied into the chamber  110  (first and/or second units e.g.) from outside placed the chamber  220 ,  230 , and others of chamber or non-chamber (second units e.g.) through the tubes by such a way mentioned-later (see  FIG. 5, 8 ). The culture solution stored the chamber  220 ,  230 , and others in a non-illustrated device installed solution stored bags or containers in a work chamber  220  or  230  etc. placed in the low temperature chamber  210  (second units e.g.). The culture solution flows into the culture vessel  125  etc. inside the chamber  110  from outside placed the culture stored chamber  220 ,  230  etc. through tubes by the pump of syringe types or/and peristaltic types etc. which is poured into the devises while the culture solution flowing out of the culture vessel  125  etc. is discharged outside the chamber  110  through tubes. The discharged culture solution is collected by a non-illustrated device installed in a work chamber  220  or  230  and others of chamber or non-chamber etc. placed outside the chamber  110 . 
     Next, with reference to  FIG. 2 , detailed description will be given of the gas concentration control unit  140 , and the tube connecting three-way between N 2  cylinder  180  and the control unit  140  as well as chamber  110 . 
     The tube  182  extending from N 2  cylinder  180  is connected to the gas concentration control unit  140  and chamber  110  via a branch coupler  183 . A manual valve  184  is installed between the branch coupler  183  and chamber  110 . When the manual valve  184  is turned on by operator, N 2  is freely supplied in large dose from N 2  cylinder  180  into the chamber  110  through sterilization filter  151  and when the manual valve  184  is off, N 2  is not freely supplied in large dose from N 2  cylinder  180  into the chamber  110 . 
     The concentration control unit  140  is equipped with predominantly-comprised that first to fourth connectors are  141  to  144  detachably connected with tubes, first to third mixing devices are  145  to  147  mixed with gas of two or more types gas, first and second electromagnetic valves are  148  and  149  adjusted gas pressure, and connector  150  is connected to the chamber  110 . The first and second mixing devices  145  and  146  are three-way selector electromagnetic valves and third mixing device  147  is a three-way junction of tubes. 
     The first connector  141  is connected with tube extending from N 2  gas generator  190  while second connector  142  is connected with tube extending from N 2  cylinder  180 . The first mixing unit  145  mixes N 2  gas received from N 2  gas generator  190  with N 2  gas received from N 2  cylinder  180  through first connector  141  and second connector  142 , respectively, and sends the gas mixture to first electromagnetic valve  148 . When the incubator  100  with a clean bench function (the first unit etc. in this system) is used as a clean bench and/or glove box, based on CO 2  and O 2  concentration measured with the sensor  161 , first electromagnetic valve  148  sends N 2  gas to third mixing unit  147  at such a pressure that N 2  concentration in the chamber  110  will become equal to a predetermined concentration. On the other hand, when the first unit etc. in this system is used as an incubator, first electromagnetic valve  148  does not send N 2  gas to third mixing unit  147 . 
     The third connector  143  is connected with tube extending from CO 2  cylinder  170 . Although not connected with any tube in  FIG. 2 , fourth connector  144  can be appropriately connected as required with CO 2  cylinder  170 , CO 2  generator, or device or cylinder adapted to supply another gas. The second mixing unit  146  mixes CO 2  gas received from CO 2  cylinder  170  through third connector  143  with the gas received through fourth connector  144  and sends the gas mixture to second electromagnetic valve  149 . Based on CO 2  concentration measured with the sensor  161 , the second electromagnetic valve  149  sends CO 2  gas to third mixing unit  147  at such a pressure that CO 2  concentration in the chamber  110  will become equal to a predetermined concentration. CO 2  concentration in the chamber  110  is determined appropriately depending on whether first unit etc. in this system is used as the clean bench and/or glove box, or used as the_incubator. 
     When the first unit etc. in this system is used as the clean bench and/or the glove box, the third mixing unit  147  mixes N 2  gas and CO 2  gas and sends the gas mixture into the chamber  110  through the connector  150  and sterilization filter  151 . Consequently, the concentrations of N 2 , CO 2 , and O 2  in the chamber  110  are kept constant. On the other hand, when the first unit etc. in this system is used as the incubator, the third mixing unit  147  sends CO 2  gas into the chamber  110  through the connector  150  and sterilization filter  151 , in the same way used as a clean bench and/or a glove box, above. Consequently, the concentrations of CO 2  and O 2 ,in the chamber  110  are kept constant. The concentration of CO 2  is possible to set from 0% to 20% and the concentration of O 2  is possible to se from 1% to 25%. That is, low O 2  partial pressure can be set as a setting condition. The concentration ranges can be selected appropriately according to the experiment. 
     Next, first elastic material  122  and third elastic material  120  will be described with reference to  FIG. 3, 5 . 
     Third elastic material  120  is made of closed-cell foam sponge, and is attached the all circumference on the inside site of near an outer edge in the chamber  110  that part of the peripheral portion  112  which is overlapped by the closed sealing door  117 , i.e. Possible flexible materials for the closed-cell foam sponge include flexible materials such as polyurethane, chloroprene, ethylene, propylene, diene, rubber etc. Third elastic material  120  includes plurality of third channels  126   a  to  126   d  making up part of a line (in/out) port. Third channels  126   a  to  126   d  extend in a width direction of third elastic material  120 , that is, in such a direction as to perforate outside and inside of the chamber  110  when the sealing door  117  is closed. Third channels  126   a  to  126   d  are installed in such a way as to be parallel to each other. 
     First elastic material  122  is made of a flexible material such as polyurethane, polyvinyl chloride, ethylene, propylene, rubber etc., and is attached on circumference of the peripheral portion  112  in the chamber  110 , and placed on the inside than third elastic material  120 . The location of first elastic material  122  is overlapped area from a slightly larger than outside of the edge in the inner door  118  to the inside edge of chamber  110  in the peripheral portion  112  by the closed inner door  118 . First elastic material  122  includes plurality of first channels  127   a  to  127   d  making up part of the line (in/out) port. First channels  127   a  to  127   d  extend in a width direction of first elastic material  122 , that is, in such a direction as to perforate outside and inside of the chamber  110  when the inner door  118  is closed. First channels  127   a  to  127   d  are installed in parallel to each other. 
     Third channels  126   a  to  126   d  and first channels  127   a  to  127   d  are installed in parallel to each other. Also, third channel  126   a  and first channel  127   a , third channel  126   b  and first channel  127   b , third channel  126   c  and first channel  127   c , and third channel  126   d  and first channel  127   d  are located on the same straight lines, respectively. 
     Next, the second elastic material  121  will be described with reference to  FIG. 4, 5 . 
     Preferably, second elastic material  121  is made of flexible material such as polyurethane, and is attached on the slightly inside circumference from outer edge in the sealing door  117 , attached on such a location as to overlap the peripheral portion  112  when the sealing door  117  and the inner door  118  are closed, i.e. Second elastic material  121  includes a plurality of second channels  128   a  to  128   d  making up part of the line (in/out) port. Second channels  128   a  to  128   d  extend in a width direction of second elastic material  121 , that is, in such a direction as to perforate outside and inside of the chamber  110  when the sealing door  117  is closed. Second channels  128   a  to  128   d  are installed in parallel to each other. 
       FIG. 5  shows the third channels  126   a  to  126   d , second channels  128   a  to  128   d , and first channels  127   a  to  127   d  through which tubes  129   a  to  129   d  are passed, with the inner door  118  and sealing door  117  closed. The tubes  129   a  to  129   d  are connected to a culture solution storing tanks or bags installed outside the chamber  110 , and are used to make a culture solution flow in and out of a petri dish or other culture devises. Note that in the vertical direction of  FIG. 5 , positional relationships among the third channels  126   a  to  126   d , second channels  128   a  to  128   d , and first channels  127   a  to  127   d  are shown in an exaggerated manner. 
     Positions of third channel  126   a , second channel  128   a , and first channel  127   a  are determined such that straight line extending along third channel  126   a , second channel  128   a , or first channel  127   a  will not cross an end of another adjacent channel. That is, the positions of the third channel  126   a , second channel  128   a , and first channel  127   a  are determined such that straight line extending along third channel  126   a  will not cross the end of the adjacent second channel  128   a , that straight line extending along second channel  128   a  will not cross the ends of the adjacent third channel  126   a  and first channel  127   a , and that straight line extending along first channel  127   a  will not cross the end of the adjacent second channel  128   a . This is also true for third channels  126   b  to  126   d , second channels  128   b  to  128   d , and first channels  127   b  to  127   d , and thus description thereof will be omitted. Consequently, the ends of third channels  126   a  to  126   d , second channels  128   a  to  128   d , and first channels  127   a  to  127   d  are not located close to each other, and thus no gas flows in and out of the chamber  110  easily through the channels. That is, inflow and leakage of gas are prevented. 
     The tube  129   a  is fitted into the third channel  126   a , second channel  128   a , and first channel  127   a  and enters the chamber  110  from outside the chamber  110 . Similarly, tubes  129   b  to  129   d  enter the chamber  110  from outside the chamber  110  through third channels  126   b  to  126   d , second channels  128   b  to  128   d , and first channels  127   b  to  127   d . Since the channels are provided in third elastic material  120 , second elastic material  121 , and first elastic material  122 , the tubes  129   a  to  129   d  enter the chamber  110  without being squeezed by third elastic material  120 , second elastic material  121 , and first elastic material  122 , allowing liquid to flow in and out of the chamber  110  easily. 
     In culturing cells put in petri dish or other culture devises etc., tubes already attached aseptically to the petri dish, or others before setting up the culture devises in the chamber  110  may have been made unremovable to maintain an aseptic condition. In this case, if third channels  126   a  to  126   d , second channels  128   a  to  128   d , and first channels  127   a  to  127   d  are used as placed with the present embodiment, the culture devises can be set up from outside to inside the chamber  110  without removing the tubes from the culture devises. 
     Next, the first hooks  114  and an operation cover  136  will be described with reference to  FIGS. 6, 11, and 12A to 12D . These figures show an example in which a glove whose fingers at the tip are not cut off is used as the operation cover  136 , i.e., a setup example in which an operation cover  136  of a single-layer structure is used as for the glove box, e.g. 
     A circular frame  130  (see  FIG. 6 ) made of silicone is attached to an inner circumferential portion of the operation cover fixing hole  119  (see  FIG. 1 ) for the operation cover  136  such as an arm cover/glove. The frame  130  includes an inner circumferential portion adapted to cover an inner circumferential surface of the operation cover fixing hole  119  and an edge portion  131  protruding slightly from a circumferential end of the inner circumferential portion in a radial direction of the operation cover fixing hole  119 . A ring of disk-shaped flange  132  protruding in the radial direction of the operation cover fixing hole  119  from the inner circumferential portion at a slight distance from the inner door  118  is attached to the edge portion  131  of the frame  130 . The flange  132  is coaxial with the frame  130  and is larger in maximum diameter than the frame  130 . 
     Next, when the chamber  110  is not used as a single type of glove box, in which an operation cover  136  of non-single-layer structure, such as the first unit e.g. is used as clean bench etc, the operation covers  136  is used of the double-layer structure in which a setup gloves whose fingers at the tip are cut off one and non cut off the other, i.e., (See  FIG. 13 ) A stopper  133  (see  FIG. 6 ) made of silicone rubber is fitted into the inner circumferential portion of the operation cover fixing hole  119 , or to put it more specifically, into an inner circumferential portion of the circular frame  130 . The stopper  133  is a disk plate shaped as a truncated cone and is inserted into the inner circumferential portion of the frame  130  with a surface with a smaller diameter foremost, bringing a circular conical surface into close contact with the inner circumferential portion of the frame  130  and thereby making it difficult for gas to flow into or out of the chamber  110  through the inner circumferential portion of the frame  130 . 
     Also, a membrane  134  is attached on the inner door  118  to cover hole of all at an outer side of the operation cover fixing hole  119  using membrane fixture plates  134   a  and  134   b . The membrane fixture plates  134   a  and  134   b  are plate materials made of metal. The membrane fixture plate  134   a  is installed above the operation cover fixing hole  119  in  FIG. 6  while the membrane fixture plate  134   b  is installed below the operation cover fixing hole  119  in  FIG. 6 . The fixture plate  134   a  and  134   b  are fixed to the inner door  118  using plural bolts and nuts, with the membrane  134  interposed between the fixture plate  134   a  or  134   b  and inner door  118 . Consequently, upper and lower sides of the membrane  134  are fixed to the inner door  118  in  FIG. 6 , but properly, the lower side of membrane fixture plate  134   b  may not use. The membrane  134  is rectangular in shape, made of a silicone or a Teflon (registered trademark) material, and provided without/with slit  135  such as an arrow-shaped etc. Any side of the membrane  134  is longer than a diameter of the operation cover fixing hole  119 . Consequently, the membrane  134  covers the entire operation cover fixing hole  119 . The slit  135  has the shape of an arrow etc. pointed downward in  FIG. 6 , perforating the membrane  134  in a thickness direction of the membrane  134 . Note that the shape of the slit  135  can be changed appropriately according to application, and the membrane  134  may be used without any slit by being fixed only by the upper fixture plate  134   a  without using the lower fixture plate  134   b . The operator can take a hand wearing a sterilized glove or take sterilized tools, samples etc. in and out of the chamber  110  through the slit  135 , or through the lower side of the membrane  134  where the membrane  134  is not fixed membrane plate  134   b  when no slit is provided. In  FIG. 3 , since the upper and lower sides of the membrane  134  are fixed to the inner door  118  with membrane fixing plate  134   a  and  134   b , in each, upper and lower, even if the hand and tools are moved in and out through the slit  135 , the membrane  134  does not cling to the hand or tools. This improves working efficiency. 
     A sterilized glove with tips thereof not cut off made of a silicone material etc., a sterilized tubular glove with tips thereof cut off, or a sterilized and/or disposable tubular operation cover  136  made of non-cloth paper or etc. is attached to the flange  132 . The end of the operation cover  136  is wound around the flange  132  and fixed to the flange  132  by being pinched by a circular fixture  300  attached to a fixed or removable tool of a glove and/or arm cover. Referring to  FIG. 11 , the circular fixture  300  is made of metal which is possible to autoclave sterilization and mainly composed of half rings ( 301 ,  302 ), a hinge  303 , and a tension spring  304 . The half rings ( 301 ,  302 ) are jointed each other at the ends via the hinge  303  so as to be pivotable relative to each other on the hinge  303 . The other ends of the half rings ( 301 ,  302 ) are linked to each other via the tension spring  304 . The tension spring  304  applies a force in such a direction that the other ends will contact each other. As the half rings ( 301 ,  302 ) pivot on the jointing hinge  303 , separating the ends of jointing spring  304  from each other, the ends of jointing spring  304  are brought back into contact with each other by being pulled by the tension spring  304 . Consequently, the half rings ( 301 ,  302 ) are held pivotally in such a way as to form an annular ring. Since the half rings ( 301 ,  302 ) are held pivotally in such a way as to form an annular ring, the operator can easily fix and remove the circular fixture  300  within the chamber  110  with one hand. 
     A technique for attaching the operation cover  136  to the operation cover fixing hole  119  will be described with reference to  FIGS. 12A to 12D . Referring to  FIG. 12A , first the operation cover  136  is passed inside the circular fixture  300 . Next, referring to  FIG. 12B , an opening of the operation cover  136  is turned inside out to cover the circular fixture  300  over an outer circumference of the circular fixture  300 . Next, referring to  FIG. 12C , the operation cover  136  with the circular fixture  300  attached thereto is put on the flange  132  while opening the circular fixture  300 . Next, when the operator takes off the hand from the circular fixture  300  referring to  FIG. 12D , the circular fixture  300  closes and gets engaged with the flange  132 . Consequently, the operation cover  136  and circular fixture  300  are attached to the operation cover fixing hole  119 . Note that the shape of the circular fixture  300  is not limited to the shown that in  FIG. 11 , and the material of the circular fixture  300  does not need to be a metal. If ethylene oxide gas sterilization, ethanol sterilization (70%), or autoclave sterilization is possible, the circular fixture  300  is possible alternatives include a string-like fixture made of silicone rubber or cloth, capable of being formed into a circular shape, and equipped with a tool such as a cord stopper used to open and close a drawstring of a pouch. The fixing end in the operation cover  136  which is attached to the flange  132  is larger than the other of non-fixing end. The operation cover  136  is preferably tapered shape toward the non-fixing end with the inside chamber  100  from the fixing side of the inner door  118 . When the incubator with a clean bench function (the first unit e.g.) is used as a clean bench, the operation cover  136  is attached to the flange  132  and the operator turn off the UV lamps and inserts a hand wearing a sterilized glove with the tip non-cut off into the operation cover  136 , such as a tubular glove with the tip cut off or an arm cover. Since the operation cover  136  is tapered as described above, the intact glove put on the hand comes into close contact with the operation cover  136 , making it difficult for gas to flow between the intact glove put on the hand and the operation cover  136 . When the chamber  110  is not used as a glove box, the non-fixing end to the around of midsection in the operation cover  136  can be left hanging on the first hook  114  etc. as take care of the non-fixing end of the operation cover be left inside the chamber  100 . When the around of midsection in the operation cover  136  is suspended on the hook etc., the operation cover  136  is bent back and forth by gravity. This makes it difficult for gas to flow into or out of the chamber  110  through the inside of the operation cover  136 . 
     The first hook  114  is made of a metal rod having a circular cross section and shaped like a letter L whose corner describes an arc. The shape of the hook may be plate-like as with a second hook  138  described later, and the L-shaped corner may be square or arc-shaped, either way. The end of the first hook  114  is attached to the inner side surface  113  using a nut  137 . The first hook  114  is located at such a position on the inner side surface as to allow the glove or operation cover  136  to be hung and held thereon, and moreover allow the operator to take with the inner glove worn on the hand, or the hand itself, out of the chamber  110  easily while leaving the operation cover  136  inside the clean bench by hanging from a neighborhood of the non-fixing end to the around of midsection in the operation cover  136  on the first hook  114  as described later. 
     A case in which an operation cover  136  of the double-layered structure is used will be described with reference to  FIG. 13 . The operation cover  136  is made up of a sheath. The sheath is made up of glove with the tips cut off or an arm cover made of non-cloth paper, cloth, silicone material, Teflon (registered trademark) material etc., and has cylindrical shape. Preferably, the cylindrical shape is tapered, decreasing in diameter from the end of fixing site toward the end of non-fixing site. The operation cover  136  is attached to the operation cover fixing hole  119  using the above-mentioned circular fixture  300 . The circular fixture  300  is attached to the end with the larger diameter. A glove  305  is inserted into an inner circumference of the operation cover  136 . The glove  305  has intact tips. As the glove  305  is inserted inside the operation cover  136 , a double-layered structure is realized. As described later, because large amount of N 2  gas flows into the chamber  110 , maintaining positive pressure inside the chamber  110 , even if the glove  305  is not put in close contact with the inner circumference of the operation cover  136 , outside air does not enter the chamber  110  from outside. 
     Next, with reference to  FIGS. 1, 2, and 6 , description will be given of the incubator equipped with the clean bench function (the first unit e.g.), used as the clean bench and as the glove box combined with the clean bench function. Here, the tubular glove with the tip cut off (installed in the operation cover fixing hole  119  of the chamber  110 ) and the glove with intact tips (put on operator&#39;s hand) are used in layers as double gloves. Also, the glove box combined with the clean bench function is used as the glove box which combines the open system and the closed system. The tubular glove with the tip cut off, for which the tubular arm cover may be used, is the operation cover  136 . 
     First, the operation cover  136  is attached to the flange  132 , and the UV lamp  116  is turned on or ethanol disinfection above half-hourly is carried out to sterilize inside the chamber  110  under the condition of closed the stopper  133 . Subsequently, CO 2  and N 2  gas are flowed in the chamber  110  controlled by the concentration control unit  140  until the stable of predetermined concentrations. Next step is slightly opened the stopper  133  and N 2  gas is flowed into the chamber  110  from the N 2  cylinder  180  by opening the manual valve  184 . Consequently, large amount of N 2  gas flow into the chamber  110 , creating positive pressure inside the chamber  110  relative to atmospheric pressure and preventing the atmosphere containing various germs from flowing into the chamber  110 . While maintaining this condition, operator turn off the UV lamp  116  and open the stopper  133 , and then takes pre-sterilized culture vessels and tools and container containing samples in and out of the chamber  110  through the lower side of the membrane  134  in disassembly the fixture plate  134   b  attached the inner door  118  or operation cover fixing hole  119  or through the slit  135  in assembly the membrane fixture plate  134   a  and  134   b , with properly ethanol disinfection around. The samples are, for example, cells etc. in dishes made of glass and more in cooling box according to need. After the above-mentioned operations, the inner door  118  and/or operation cover fixing hole  119  is closed, the UV lamp  116  is turned on, the manual valve  184  and the stopper  133  are properly closed, and the inside of the chamber  110  going through preparatory work is sterilized. The necessity of the case is taken the samples into the chamber  110  after the above preparatory work in sterilized. Sterilized tubes used to supply and discharge the culture solution to/from the culture devise during the preparatory work are laid through the channels of the elastic materials, first  127   a - 127   d  etc and second  128   a ,- 128   d  etc and third  126   a - 126   d  etc, between the inner door  118  or the sealing door  117  and the chamber  110  for the passes through the inside from/to outside the chamber  110  (see  FIG. 5 ). At the time of work, operator opens the manual valve  184  and the stopper  133 , turns off the UV lamp  116 , inserts the hand wearing the sterilized glove with intact tips into the operation cover  136 , and manipulates the samples, the culture medium etc. When the manipulations are finished, operator closes the gloved hand into a fist such that glove fingers which touched the culture medium, samples etc. will be placed inside the palm of the glove worn on the hand. And then, operator hangs from a neighborhood of the non-fixing end to the around of midsection in the operation cover  136  on the hook  114  by moving the arm and elbow, with the back of the up, in such a way that fingers will not touch the hook  114 . The next step, operator pulls the hand together with the glove out of the operation cover  136  while taking care inside its ( 136 ) circumference of no-touch with the glove fingers grasping hands which touched the culture medium, samples etc. Then, after the exchange of the glove worn on the hand with a new sterilized glove outside the chamber  110 , operator closes the operation cover fixing hole  119  with the stopper  133  properly sterilized with ethanol (70%), turns on the UV lamp  116 , and closes the manual valve  184 . 
     At the time of clean bench work, as above-mentioned, just before starting the work, with a large amount of N 2  gas allowed to flow into the chamber  110  by opening the manual valve  184 , operator takes pre-sterilization of the culture vessel and the tools and container containing samples in and out mainly through the lower side of the membrane  134  with free from the fixture plate  134   b  attached the operation cover fixing hole  119  or through the slit  135  while minimizing the duration, opening degree, and frequency of opening the inner door  118 . Consequently, changes in the concentrations and temperatures of O 2  and CO 2  in the chamber  110  being used as a clean bench are minimized. 
     For the non-impairment of function in the clean bench, it is honked or alarm of warning sound in the cases of deviation from the range of advance preparation setting on each O 2  or CO 2  concentration in this system. In the cases of honked of warning sound or the ringing on ahead, the stopper  133  is opened/closed or the change of opening degree in the stopper  133  with rotation angles among the operation caver fixing hole  119  is controlled by the breaking through gas between the stopper  133  and the operation cover fixing hole  119 , while opening the manual valve  184  and with properly repeating above sterilization. 
     For the safety and protect for operator against infection etc., the time on working inside the chamber  110  or after-mentioned chamber  220 ,  230  in this system is essential that UV lamp  116  turn off and operator wears a mask etc. and sterilized disposable and non-permeability of gloves for the non-contact with samples or culture buffer and so on. 
     Regarding how to hold the vessel/container when taking in and out sterilized the culture vessel or the tools and container containing samples, similarly the vessel/container is taken in and out by being held with the gloved hand closed into a fist with the fingers of the glove worn on the hand turned inward or the vessel/container is set up inside the chamber  110  through the operation cover  136  via the lower side of the membrane  134  or via the slit  135  by holding the vessel/container or tool from above, with the back of the hand describing an arc such as when a container such as a tea caddy for powdered green tea is held during tea ceremony. Also, in taking a tool or vessel/container out of the chamber  110  together with the glove worn on the hand, the neighborhood of the end of the operation cover  136  is hung on the hook  114  or the like by moving the arm and elbow without using fingertips and the work is carried out in a similar manner. 
     Also, in pulling the glove worn on the hand out of the chamber  110  at the end of a culture operation etc., if the glove fingers which touched the culture medium, samples, or are fear in probability to have touched, an inner side of an operation cover  136 , in the case of long-term culture, the operator carries out disinfection by turning on the UV lamp  116  not less than a half-time with closed the manual valve  184  and the stopper  133 . And then, operator wears a new sterilized glove and mask outside the chamber  110 , leaves in the membrane  134  and opens the manual valve  184  and the stopper  133  again, and turns off the UV lamp  116 , inserts the hand wearing the new sterilized glove into the chamber through the operation cover fixing hole  119  under the lower side of the membrane  134  for the non-polluted operation cover  136  (on one side) which did not touch the fingers of glove which had touched the culture medium, samples etc. In next, operator takes off the polluted operation cover  136  (on the other side) with the unopened inside polluted site in which touched the fingers of glove that had touched the culture medium, samples etc., from the circular fixture frame  130  include the flange  132 , and speedily encloses its in using sterilization bags, and exchange the new operation cover  136  with a sterilized and prepared beforehand inside the chamber  110 . After the change of this, operator turn on the UV lamp  116  and closed the stopper  133  and the manual valve  184 . Note that if a contaminated glove touches the membrane  134  or the slit  135  in the membrane  134 , similarly the membrane  134  can be easily replaced with a sterilized new membrane  134  by taking off the fixture plate  134   a . The present embodiment not only makes it easy to exchange gloves while carrying out work or continuing culture, which is difficult with the conventional glove box, but also makes it possible to exchange of all commodities, such as the membrane  134  and operation cover  136  etc., on any route through which tools and samples containers etc. pass from the inside of the chamber  110  to the outside of the inner door. 
     What has been described above is only an example, and depending on culture conditions, the tubes used to supply and discharge culture solutions may become unnecessary; and under conditions in which O 2  and CO 2  concentrations are close to atmospheric pressure or during work in which things are taken in and out of the box for a shorter duration or less frequently, the operation cover  136  is unnecessary, and the membrane  134  and slit  135  provided in the operation cover fixing hole  119  and inflow of large amount of N 2  gas into the box  110  caused by opening the manual valve  184  may be sufficient. 
     Next, with reference to  FIGS. 1, 2, and 6 , description will be given of the incubator  100  equipped with the clean bench function (the first unit e.g.) used as the glove box. Here, description will be given of the embodiment which uses the single-glove with intact tips and uses this system of function as the closed-system glove box which does not require gloves and tools to be exchanged or replenished during culture. Specifically, it is the case of non-using the function of clean bench in culturing stage, exclude of preparation stage etc., and so the manual valve  184  and the inner door  118  and the stopper  133  etc. are all closed in culturing stage. 
     First, the glove with intact tips is attached as an operation cover  136  to the operation cover fixing hole  119  via the flange  132 , UV lamp  116  is turned on, and ethanol disinfection is carried out to sterilize inside the chamber  110 . The manual valve  184  is opened, allowing large amount of N 2  gas to flow into the chamber  110  from N 2  cylinder  180 , and then the stopper  133  is slightly opened. Next, UV lamp  116  is turned off, and then culture vessels and tools and samples are carried and placed in the chamber  110  for preparation of culture. In the culture stage, the inner door  118  or the stopper  133  of the operation cover fixing hole  119  is closed, the UV lamp  116  is turned on if objects to be cultured is not affected, and the manual valve  184  is closed. In the case of apprehensive affected objects by UV lamp  116 , it is necessary is placed on the samples under the foil-wrapped cover or container, or properly turn off the UV lamp  116 . When the culture vessels and tools and samples are set up in preparation, tubes used to supply and discharge the culture solution to/from the samples are laid through the channels of the elastic materials, first  127   a - 127   d  etc and second  128   a ,- 128   d  etc and third  126   a - 126   d  etc, between the inner door  118  or the sealing door  117  and the chamber  110  for the passes through the inside from/to outside the chamber  110  (see  FIG. 5 ). If it is not necessary to perfuse the culture medium, the pipes used to supply and discharge a culture solution may not be laid. 
     When the inner door  118  is opened or closed before the start of culture or after the end of culture, if the duration, opening degree, and frequency of opening/closing the inner door  118  are minimized with large amount of N 2  gas being allowed to flow into the chamber  110  by opening the manual valve  184 , the temperatures and concentrations of O 2  and CO 2  are maintained as described above. While maintaining this condition, the operator carries out work with the UV lamp  116  turned off during the work. In the treatment of warning samples and culture medium and materials etc. that suspected to affect human bodies by substances of harmful bacteria or toxic reagents etc. in which desired to be avoided flowing out of the chamber  110  in vapor phase and others, note that if the samples or the reagents is taken out of an enclosed airtight container in the chamber  110 , the manual valve  184  is kept closed, and the pump  154  etc. which for vapor phase exchange between the first work chamber  220  described later and the chamber  110  is switched off and shut down. Operator works inside the chamber  110  from gloves  136  with intact tips on the operation cover fixing holes  119 , and after the work, operator put out the hands to keep the gloves inside the chamber  110  and UV lamp  116  is turned on to disinfect the inside the chamber  110 . In the work beginning in preparation, there are doing the operation of setting up the container with the sample and reagent enclosed therein in the chamber  110 , and laying the tubes used to supply and discharge the culture solution in/from the inner door  118 , and ending with the operation of sterilizing the exterior of the container inside the box with UV lamp  116 , which is similar to the work described above. 
     Consequently, the exchange of the culture solution can be supplied by perfusion in/out the culture solution while maintaining the culture condition including, temperatures, O 2  and CO 2  concentration etc. and the culture solution can be supplied from outside system  100  without opening/closing the inner door  118  or the operation cover fixing hole  119  in the present system  100 . For the safety and protect of operator against infection etc., and for the care of honked or alarm of warning sound on gas control, it is described as the preceding paragraph &lt; 0047 &gt;. 
     Furthermore, if there is concern about safety of the operator, the sample can be manipulated using the double-glove structure made of gloves with intact tips by inserting a hand wearing the sterilized glove with intact tips into the glove  136  with intact tips, the glove  136  being fixed to the chamber  110 . It is possible to use this system for globe box in non-using the function of clean bench not only culturing stage but also preparation and cleanup after the culture. 
     Next, with reference to  FIGS. 1, 2, and 6 , description will be used as the incubator function instead of non-using clean bench function in this system of the incubator  100  equipped with a clean bench (the first unit e.g.). In this case, samples manipulated using the clean bench function and glove box function is already placed in the chamber  110  and the inner door  118  is closed. In this condition, the inner circumferential stopper  133  has been fitted in the operation cover fixing hole  119  and the sealing door  117  has been closed. At this time, tubes used to supply and discharge a culture solution to/from the samples have been laid among third channels  126   a  to  126   d , second channels  128   a  to  128   d , and first channels  127   a  to  127   d  (see  FIG. 5 ). The temperatures and O 2  concentration and CO 2  concentration are maintained constant as described above by making N 2  gas and CO 2  gas flow into the chamber  110  as required. While maintaining this condition, the culture solution is supplied to the sample through the tubes to culture the samples. Regarding the condition during culture, the inside of the culture vessel is observed from outside the transparent inner door  118  using a microscope with a variable observation angle with the culture vessel remaining set up inside the chamber  110 . Since microscopic observation can be conducted without opening the inner door  118  or the stopper  133  of the operation cover fixing hole  119 , even for consecutive observation during culture, there is no need to take the culture vessel out of the incubator chamber  100 , the temperatures and concentrations of O 2  and CO 2  are maintained constant, and there is less stress on the culture samples. Alternatively, it is also possible to observe the condition of culture in the culture vessel inside the chamber  110  by inserting a microscope objective lens shaped like a fiber cable etc. used for endoscopy etc. through the channels  126  in sealing materials between inner door  118 , and through the lower side of the membrane  134  on free from the membrane fixing plate  134   b  attached on the outside of the operation cover fixing hole  119  or through the slit  135  via the operation cover  136  attached to the flange  132  by simultaneously combination using the clean bench function of allowing large amount of N 2  gas to flow into the chamber  110  from N 2  cylinder  180  by opening the manual valve  184  and slightly opening the stopper  133  in adjusted time. 
     Next, the second hook  138  having another shape will be described with reference to  FIG. 7 . The second hook  138  is made of plate-like metal and shaped like a letter S whose corners are square. The hook has the same uses as the first hook  114 , and thus may have a shape similar to that of the first hook  114 . The end of one side in the second hook  138  is attached to the inner side surface  113  using the bolt  139 . The position of the second hook  138  on the inner side surface  113  is similar to that of the first hook  114 , and thus description thereof will be omitted. 
     According to the present embodiment, after being manipulated, cells etc. can be cultured without moving the culture vessel  125  out of a clean bench, i.e., out of the chamber  110 . Also, the culture environment which satisfies the low-temperature or low O 2  condition can be created easily. 
     Next, an incubator system  200  (the second unit e.g.) according to the second embodiment will be described with reference to  FIGS. 8 to 10 . Components similar to those of the first embodiment in the first unit e.g. are denoted by the same reference numerals as the corresponding components, and description thereof will be omitted. The present embodiment in the second unit e.g. differs from the first embodiment in that the incubator system  200  further includes a low temperature chamber  210 , the first work chamber  220 , the second work chamber  230 , and gas control unit  240 . Mainly these components will be described below. 
       FIG. 8  is a schematic diagram of the incubator system  200  (the second units e.g.). The incubator system  200  mainly includes the incubator  100  with a clean bench function (the first and/or second unit in this system), the low temperature chamber  210  (the second unit e.g.), the first work chamber  220  (the second unit e.g.), the second work chamber  230  (the second unit e.g.), and the gas control unit  240  (the second unit e.g.). Note that for the sake of explanation, the chamber  110  (the first and/or second unit e.g,), low temperature chamber  210 , first work chamber  220 , and second work chamber  230  are illustrated in such a way as to allow inside these components to be seen through. 
     The low temperature chamber  210  has the shape of rectangular parallelepiped and includes left-hand (which in possible of right-hand, too) front door  211  attached to the front of the low temperature chamber  210  and made of transparent polycarbonate plate, heater, and refrigerating machine  213 . The heater and refrigerating machine  213  maintains predetermined temperatures inside the low temperature chamber  210 , e.g., temperatures in the range from 4 degree Celsius in air temperature to 50 degrees Celsius adding in room temperature, based on the temperature detected by a non-illustrated temperature sensor attached inside the low temperature chamber  210 . The refrigerating machine is used, for example, for a range from 4 degrees Celsius in air temperature (inclusive) to a neighborhood of room temperature, and the heater is used, for example, for a range from a neighborhood of room temperature to 50 degrees Celsius adding in room temperature (inclusive). The room temperature is, for example, from 25 to 27 degrees Celsius. One or more shelf boards  212  are installed inside the low temperature chamber  210 . The first and second work chambers  220  and  230  are possible to set up on the shelf board(s)  212 . 
     The first work chamber  220  has the shape of a rectangular parallelepiped and mainly includes the opening  261  formed by opening in the front of the first work chamber  220 , the right-hand (which in possible of right-hand, too) front door  225  attached to the opening  261  using pivotable hinges, the UV lamp  226 , and the sensor  269 . The front door  225  is made of flat transparent polycarbonate plate about large enough to completely close the opening  261  and provided with the rectangular hole  228  perforating the polycarbonate plate in the thickness direction below the vertical center and two arm cover/glove fixing holes (the operation cover fixing hole)  229  perforating the polycarbonate plate in the thickness direction above the vertical center. The rectangular hole  228  has a width and height about large enough to pass the culture vessel  125 . The upward-opening rectangular door  227  is attached to the rectangular hole  228 . The rectangular door  227  is provided with non-illustrated locking mechanism to prevent the rectangular door  227  from opening inadvertently. The sensor  269  includes temperature sensor, CO 2  sensor, and O 2  sensor, and measures temperature, CO 2  concentration, and O 2  concentration in the first work chamber  220 . 
     The configuration of the two operation cover fixing holes  229  is similar to that of the operation cover fixing holes  119  according to the first embodiment, and thus description thereof will be omitted. The UV lamp  226  is attached to the inner back surface of upper site in the first work chamber  220  and emits UV rays whose wavelength is suitable for sterilization. The hook similar to the first hook  114  or second hook  138  is attached and plural shelf boards  265  (see  FIG. 9 ) can be attached inside the first work chamber  220 . Configurations of the first hook  114 , second hook  138 , UV lamp  226 , and shelf board  265  are similar to those of the first embodiment, and thus description thereof will be omitted. Also, the first hook  114  or second hook  138  is omitted in  FIG. 9  for simplicity of explanation. The non-illustrated heat insulating material is attached the ceiling surface, an inner back surface, an inner side surface, and the bottom surface of the first work chamber  220 . Fourth elastic material  263  and fifth elastic material  264  (see  FIG. 9 ) are attached that part of peripheral portion  262  which is overlapped by the closed front door  211 . 
     The first work chamber  220  mainly includes seventh to ninth connectors  221  to  223  detachably connected with tubes, fifth electromagnetic valve  224  adapted to adjust gas pressure, and seventh mixing unit  268  adapted to mix plural types of gas. Seventh mixing unit  268  is three-way junction tube. Operations of these components will be described later. 
     Next, fourth elastic material  263  and fifth elastic material  264  will be described with reference to  FIG. 9 . 
     Fourth elastic material  263  is preferably made of closed-cell foam sponge of flexible material such as consist of polyurethane, chloroprene, ethylene, propylene, diene, rubber etc. and is attached on the entire outer edge in the chamber  220  that part of the peripheral portion  262  which is overlapped by the closed front door  225 . Fourth elastic material  263  includes plurality of fourth channels  266   a  to  266   d  making up part of the line (in/out) port. Fourth channels  266   a  to  266   d  extend in width direction of fourth elastic material  263 , that is, in such a direction as to perforate outside and inside of the first work chamber  220  when the front door  225  is closed. Fourth channels  266   a  to  266   d  are installed in such way as to be parallel to each other. 
     Fifth elastic material  264  is preferably made of the flexible material such as consist of polyurethane, vinyl chloride, ethylene, propylene, rubber etc. and is attached on the entire inner edge in the chamber  220  that part of the peripheral portion  262  which is overlapped by the closed front door  225 . Fifth elastic material  264  includes a plurality of fifth channels  267   a  to  267   d  making up part of the line (in/out) port. Fifth channels  267   a  to  267   d  extend in width direction of fifth elastic material  264 , that is, in such a direction as to perforate outside and inside of the first work chamber  220  when the front door  225  is closed. Fifth channels  267   a  to  267   d  are installed in parallel to each other. The fourth channels  266   a  to  266   d  and fifth channels  267   a  to  267   d  are installed in parallel to each other. Also, fourth channel  266   a  and fifth channel  267   a , fourth channel  266   b  and fifth channel  267   b , fourth channel  266   c  and fifth channel  267   c , and fourth channel  266   d  and fifth channel  267   d  are located on different straight lines, respectively. Consequently, ends of fourth channels  266   a  to  266   d  and fifth channels  267   a  to  267   d  are not located close to each other, and thus no gas flows in and out of the first work chamber  220  easily through the channels. That is, inflow and leakage of gas are prevented. 
     The second work chamber  230  mainly includes an upward-opening front door  235  and a UV lamp  236 , where the front door  235  includes a rectangular hole  238 , two operation cover fixing holes  239 , and a sensor  279 . The configuration of the second work chamber  230  is similar to that of the first work chamber  220 , and thus description thereof will be omitted. The second work chamber  230  mainly includes 20th to 22nd connectors  231  to  233  detachably connected with tubes, sixth electromagnetic valve  234  adapted to adjust gas pressure, and an eighth mixing unit  278  adapted to mix plural types of gas. Eighth mixing unit  278  is a three-way junction tubes. Operations of these components will be described later. The sensor  279  includes temperature sensor, CO 2  sensor, and O 2  sensor, and measures temperature, CO 2  and O 2  concentration in the second work chamber  230 . 
     Next, the tubes used to connect various materials as well as the gas control unit  240  will be described in detail with reference to  FIG. 10 . 
     The tube  182  extending from N 2  cylinder  180  is connected to the gas control unit  240  via branch couplers  185 ,  183 , and  186 ; connected to the first work chamber  220  and second work chamber  230  via branch couplers  185  and  214 ; connected to the chamber  110  via branch couplers  185  and  183 ; and connected to the concentration control unit  140  via the branch couplers  185 ,  183 , and  186 . 
     The tube  192  extending from N 2  gas generator  190  is connected to the concentration control unit  140  and gas control unit  240  via a branch coupler  191 . 
     The tube  172  extending from CO 2  cylinder  170  is connected to the concentration control unit  140  and gas control unit  240  via a branch coupler  173 . 
     The concentration control unit  140  mainly includes fifth and sixth connectors  152  and  153  detachably connected with tubes, the first pump  154 , and seventh and eighth connectors  155  and  156  adapted to interconnect the first pump  154  and chamber  110 . 
     Fifth and sixth connectors  152  and  153  are connected with tubes extending from the first work chamber  220 . First pump  154  aspirates gas out of the first work chamber  220  via ninth connector  223  and fifth connector  152  and sends out the sucked gas into the chamber  110  via the eighth connector  156 . On the other hand, the first pump  154  sucks gas out of the chamber  110  via seventh connector  155  and sends out the sucked gas into the first work chamber  220  via sixth connector  153 . In the first work chamber  220 , the 18th connector  222  receives gas from the first pump  154  and sends the gas to seventh mixing unit  268 . Seventh mixing unit  268  sends the gas into the first work chamber  220  through the sterilization filter  151 . Consequently, the first work chamber  220  contains the same gas composition as in the chamber  110 . When the first work chamber  220  is used as clean bench or glove box, N 2  gas is further supplied to the first work chamber  220  from N 2  cylinder  180  via the branch couplers  185  and  214 . In the first work chamber  220 , the 17th connector  221  receives N 2  gas from N 2  cylinder  180  and sends N 2  gas to the fifth electromagnetic valve  224 . Based on CO 2  concentration and O 2  concentration measured with the sensor  269  installed inside the first work chamber  220 , fifth electromagnetic valve  224  sends N 2  gas to the seventh mixing unit  268  at such a pressure that N 2  concentration in the first work chamber  220  will become equal to the predetermined concentration. Seventh mixing unit  268  sends out N 2  gas into the first work chamber  220  through the sterilization filter  151 . This creates the positive pressure inside the first work chamber  220  relative to atmospheric pressure and thereby prevents the atmosphere containing various germs from flowing into the first work chamber  220 . 
     The gas control unit  240  mainly includes ninth to twelfth connectors  241  to  244  detachably connected with tubes, fourth to sixth mixing units  245  to  247  adapted to mix plural types of gas, third and fourth electromagnetic valves  248  and  249  adapted to adjust gas pressure, second pump  250 , the subtank  256  adapted to accumulate gas, thirteenth and fourteenth connectors  251  and  252  adapted to connect the second pump  250  to the second work chamber  230 , 15th and 16th connectors  254  and  255  adapted to connect the second pump  250  to the subtank  256 . The fourth and fifth mixing units  245  and  246  are three-way selector electromagnetic valves and the sixth mixing unit  247  is the three-way junction tube. A sensor  257  is attached in the subtank  256 . The sensor  257  includes CO 2  sensor and O 2  sensor and measures CO 2  and O 2  concentration in the subtank  256 . 
     Ninth connector  241  is connected with tube extending from N 2  gas generator  190  and tenth connector  242  is connected with tube extending from N 2  cylinder  180 . Fourth mixing unit  245  mixes N 2  gas received from N 2  gas generator  190  with N 2  gas received from N 2  cylinder  180  through ninth connector  241  and tenth connector  242 , respectively, and sends the gas mixture to the third electromagnetic valve  248 . When the second work chamber  230  is used as the clean bench or a glove box, based on the CO 2  concentration and O 2  concentration measured with the sensor  279  installed inside the second work chamber  230 , third electromagnetic valve  248  sends N 2  gas to the sixth mixing unit  247  at such a pressure that N 2  concentration in the second work chamber  230  will become equal to the predetermined concentration. The N 2  cylinder  180  supplies N 2  gas to the second work chamber  230  via the branch coupler  185 , branch coupler  214 , and twentieth connector  231 . In the second work chamber  230 , the twentieth connector  231  receives N 2  gas and sends N 2  gas to sixth electromagnetic valve  234 . Based on CO 2  and O 2  concentration measured with the sensor  279  installed inside the second work chamber  230 , the sixth electromagnetic valve  234  sends N 2  gas to the eighth mixing unit  278  at such a pressure that N 2  concentration in the second work chamber  230  will become equal to the predetermined concentration. Eighth mixing unit  278  sends out N 2  gas into the second work chamber  230  through the sterilization filter  151 . This creates positive pressure inside the second work chamber  230  relative to atmospheric pressure and thereby prevents the atmosphere containing various germs from flowing into the second work chamber  230 . On the other hand, when the second work chamber  230  is used as an incubator, third electromagnetic valve  248  does not send N 2  gas to sixth mixing unit  247 . 
     The 11th connector  243  is connected with tube extending from CO 2  cylinder  170 . Although not connected with any tubes in  FIG. 10 , twelfth connector  244  can be appropriately connected as required with CO 2  cylinder  170 , CO 2  generator, or device or cylinder adapted to supply another gas. The fifth mixing unit  246  mixes CO 2  gas received from CO 2  cylinder  170  through the 11th connector  243  with the gas received through the twelfth connector  244  and sends the gas mixture to the fourth electromagnetic valve  249 . Based on CO 2  concentration measured with the sensor  279  installed inside the second work chamber  230 , the fourth electromagnetic valve  249  sends CO 2  gas to the sixth mixing unit  247  at such a pressure that CO 2  concentration in the second work chamber  230  will become equal to the predetermined concentration. The CO 2  concentration in the second work chamber  230  is determined appropriately depending on whether the incubator  100  with a clean bench function is used as the clean bench or the glove box, or used as the incubator. 
     When the second work chamber  230  is used as the clean bench or the glove box, the sixth mixing unit  247  mixes N 2  gas and CO 2  gas and sends the gas mixture to branch coupler  253 . 
     The second pump  250  sucks gas out of the second work chamber  230  via the 22nd connector  233  and 13th connector  251  and sends out the sucked gas into the subtank  256  via the 15th connector  254 . On the other hand, the second pump  250  sucks gas out of the subtank  256  via the 16th connector  255  and sends out the sucked gas to the branch coupler  253 . At this time, the gases mixed by the sixth mixing unit  247  have been sent out to the branch coupler  253 . Consequently, the gas mixed by the sixth mixing unit  247  and the gas sucked out of the gas subtank  256  are mixed by the branch coupler  253  and sent to the second work chamber  230  via the 14th connector  252  and 21st connector  232 . The second work chamber  230  sends the gas received by the 21st connector  232  to the eighth mixing unit  278 . The eighth mixing unit  278  mixes N 2  gas received from the sixth electromagnetic valve  234  with the gas received from the 21st connector  232  and sends the gas mixture into the second work chamber  230  through the sterilization filter  151 . 
     Consequently, when the second work chamber  230  is used as the clean bench or the glove box, the concentrations of N 2 , CO 2 , and O 2  in the second work chamber  230  are kept constant and positive pressure is created inside the second work chamber  230  relative to atmospheric pressure, thereby preventing the atmosphere containing various germs from flowing into the second work chamber  230 . 
     On the other hand, when the second work chamber  230  is used as the incubator, the sixth mixing unit  247  sends CO 2  gas into the second work chamber  230  via the branch coupler  253 . Consequently, the concentrations of CO 2  and O 2  in the second work chamber  230  are kept constant. The concentration of CO 2  is capable of setting up 0% to 20% and the concentration of O 2  is capable of setting up 1% to 25%. The concentration ranges can be selected appropriately according to the experiment. 
     The present embodiment provides the first work chamber  220  containing the gas of the same composition as in the chamber  110 . Also, the present embodiment provides the second work chamber  230  containing the gas of composition different from that in the chamber  110 . 
     By saving the culture medium in the second work chamber  230  and manipulating and culturing cells in the first work chamber  220 , it is possible to save the culture medium and manipulate and culture the cells while keeping the culture medium sterile. 
     Note that the position to which the UV lamp  116  is attached is not limited to the inner back surface  115 . The position suitable for sterilizing the interiors of the chamber  110 , first work chamber  220 , and second work chamber  230  can be selected appropriately. 
     Note that the temperature inside the chamber  110  is not limited to the range from 4 degrees Celsius in air temperature to 50 degrees Celsius adding in room temperature, and may be selected appropriately according to the experiment. 
     Note that only N 2  cylinder  180  may be used without using N 2  gas generator  190 . Also, although description has been given of the configuration in which N 2  gas and CO 2  gas are supplied into the chamber  110  and controlled, only one type of gas or more and another additional gas, such as O 2  gas etc., each experiment desired additional type of gas may be supplied into the chamber  110  and controlled. In that case, the connector, the mixing unit, and the electromagnetic valve are installed appropriately. 
     Note that the third channels  126   a  to  126   d , second channels  128   a  to  128   d , and first channels  127   a  to  127   d  do not need to have substantial widths such as shown in  FIGS. 3 to 5  and may be shaped as narrow so-called slits. Also, the extending directions of the third channels  126   a  to  126   d , second channels  128   a  to  128   d , and first channels  127   a  to  127   d  are not limited to the directions described above, as long as the directions are parallel to the front of the inner door  118  or sealing door  117  and cross a pivot axis of the inner door  118  or sealing door  117 . Also, third channels  126   a  to  126   d  do not have to be parallel to each other, and similarly second channels  128   a  to  128   d  do not have to be parallel to one another, and first channels  127   a  to  127   d  do not have to be parallel to each other. Furthermore, third channels  126   a  to  126   d , second channels  128   a  to  128   d , and first channels  127   a  to  127   d  do not have to be parallel to each other. 
     Third channel  126   a  and first channel  127   a , third channel  126   b  and first channel  127   b , third channel  126   c  and first channel  127   c , and third channel  126   d  and first channel  127   d  may be located on different straight lines, respectively. 
     Also, regarding the positional relationships among third channels  126   a  to  126   d , second channels  128   a  to  128   d , and first channels  127   a  to  127   d  in the vertical direction of  FIG. 5 , the channels may be placed at closer positions than shown in  FIG. 5 . Then, the tubes  129   a  to  129   d  will be bent more gently, allowing the culture solution to flow easily in the tubes  129   a  to  129   d . Furthermore, in  FIG. 5 , the tubes  129   a  to  129   d  may be installed in third channels  126   a  to  126   d , second channels  128   a  to  128   d , and first channels  127   a  to  127   d  in a staircase pattern. That is, for example, the tube  129   d  is installed in third channel  126   d , second channel  128   d , and first channel  127   c . Consequently, the tubes  129   a  to  129   d  are bent gently, allowing the culture solution to flow easily in the tubes  129   a  to  129   d.    
     Note that fourth channels  266   a  to  266   d  and fifth channels  267   a  to  267   d  do not need to have substantial width such as shown in  FIG. 9 , and may be shaped as narrow so-called slits. Also, the extending directions of fourth channels  266   a  to  266   d  and fifth channels  267   a  to  267   d  are not limited to the directions described above, as long as the directions are parallel to the front of the front doors  225  and  235  and cross pivot axes of the front doors  225  and  235 . Also, fourth channels  266   a  to  266   d  do not have to be parallel to each other and similarly fifth channels  267   a  to  267   d  do not have to be parallel to each other. Furthermore, fourth channels  266   a  to  266   d  and fifth channels  267   a  to  267   d  do not have to be parallel to each other. 
     Fourth channel  266   a  and fifth channel  267   a , fourth channel  266   b  and fifth channel  267   b , fourth channel  266   c  and fifth channel  267   c , and fourth channel  266   d  and fifth channel  267  may be located on the same straight lines, respectively. 
     The positions of first channels  127   a  to  127   d , second channels  128   a  to  128   d , third channels  126   a  to  126   d , fourth channels  266   a  to  266   d , and fifth channels  267   a  to  267   d  are not limited to the positions described above, and may be provided on entire peripheries of first elastic material  122 , second elastic material  121 , third elastic material  120 , fourth elastic material  263 , and fifth elastic material  264 . 
     Note that the positions of the operation cover fixing holes  119  are not limited to positions below the vertical center of the inner door  118  and the positions of the operation cover fixing holes  229  and  239  are not limited to positions above the vertical center of the inner door  118 . 
     The culture vessel  125  may be test tube, tapper, bottle, petri dish, flask, or another container of any of various shapes. Although the closed-system culture method using the perfusate has mainly been described in the present embodiment, the present device is also applicable to the culture method which does not use the perfusate, the culture method which uses the open system, and the culture method which uses combination of the open system and the closed system. Also, in these culture method, the shape of culture vessel is not limited to the one described above. 
     The sensor  161  may further include N 2  sensor to measure N 2  concentration in the chamber  110 . When the incubator  100  with the clean bench function is used as the clean bench or the glove box, based on N 2  concentration measured with the sensor  161 , the first electromagnetic valve  148  may send N 2  gas to the third mixing unit  147  at such a pressure that N 2  concentration in the chamber  110  will become equal to the predetermined concentration. 
     Note that the sensor  269  may further include N 2  sensor to measure N 2  concentration in the first work chamber  220 . Based on N 2  concentration measured with the sensor  269  installed inside the first work chamber  220 , fifth electromagnetic valve  224  may send N 2  gas to seventh mixing unit  268  at such a pressure that N 2  concentration in the first work chamber  220  will become equal to a predetermined concentration. 
     Note that the sensor  279  may further include N 2  sensor to measure N 2  concentration in the second work chamber  230 . Based on N 2  concentration measured with the sensor  279  installed inside the second work chamber  230 , the third electromagnetic valve  248  may send N 2  gas to sixth mixing unit  247  at such a pressure that N 2  concentration in the second work chamber  230  will become equal to a predetermined concentration. Also, based on N 2  concentration measured with the sensor  279  installed inside the second work chamber  230 , sixth electromagnetic valve  234  may send N 2  gas to eighth mixing unit  278  at such a pressure that N 2  concentration in the second work chamber  230  will become equal to the predetermined concentration. 
     Note that the materials of the components described above are not limited to those described above. 
     The operation cover  136 , which has been or can be sterilized, is preferably the glove with intact tips, the tubular glove with the tip cut off, or the arm cover made of non-cloth paper, cloth, silicone, etc. The tubular glove with the tip cut off and the arm cover are preferably tapered in the chamber, but are possible to be otherwise tapered. The operation cover made of non-cloth paper, cloth, silicone, etc. may be shaped such that one tip and/or both tips of the tube, or midsection of the tube will be squeezed by rubber etc. 
     Note that if a strictly sterilized condition is not required, the operator may insert a bare hand disinfected with ethanol etc. into the chamber using only the sheath as the operation cover  136  without using the glove  305  on the inner side. Furthermore, if the incubator function is unnecessary or if conditions close to atmospheric pressure are enough, the case of only clean bench function of allowing flow with large amount of N 2  gas into the chamber  110  and thereby creating positive pressure inside the chamber  110  may be used without using the operation cover  136  either. 
     Note that the numbers which represent the sizes of various materials shown in the present specification and the drawings are exemplary and not restrictive. 
     Whereas embodiments of the present invention have been described with reference to the accompanying drawings, it will be apparent to those skilled in the art that changes can be made to structures and relationships of various parts without departing from the spirit and scope of the invention set forth in the appended claims.