Abstract:
A micro IVF chamber that enables rapid filling, hermetic sealing, and custom gas environment control for secure cell extraction, fertilization, culturing, insemination, macro and microscopic content examination, post examination rapid refilling, return to optimal gas levels and pH values, with direct view of cell in conventional microscopes while maintaining culture environment, designed for use in any incubator and multiple devices per shelf for optimum efficiency while maintaining integrity of individual devices and patient-culture identity in a simple one-sample clamp-and-go device is disclosed.

Description:
BACKGROUND OF THE INVENTION 
       [0001]    1. Field of the Invention 
         [0002]    The present invention relates to a device for providing a sealed and gassed and thermally stable culturing environment in a secure compact portable chamber for In Vitro Fertilization (IVF). 
         [0003]    2. Discussion of the Related Art 
         [0004]    In IVF, eggs are extracted from a woman and combined in a laboratory with sperm from a man. Fertilization usually takes place in one day. The resulting embryos are kept in an incubator for three to five days, after which time they are checked for appropriate development, and then transferred into the woman&#39;s uterus. 
         [0005]    On the day before the oocyte (egg) retrieval, oocyte culture dishes are labeled with the patients name, date of birth, and lab identification number. The number and size of the ovarian follicles determines the number of dishes prepared (one dish for each large follicle). The dishes are filled with a rinsing fluid in an outer well for removing blood and other extraneous cells from the egg, and a nutrient fluid for the culture of the egg is in a center well. 
         [0006]    The male will be asked to collect a semen sample after the oocyte aspiration. The andrologist escorting him to the collection room will identify the patient&#39;s partner from his identification bracelet prior to collection and will write both names on the outside of the specimen container after collection. The partner will be asked to verify that both names are correct. 
         [0007]    The embryologist will determine the number of eggs obtained and classify them as mature, immature, post mature or degenerative. The sperm is then processed in an IVF laboratory. Sperm and eggs are placed in the dishes and fertilization takes place (in vitro) outside the patient&#39;s body in the incubator. 
         [0008]    The dishes placed in an embryology incubator are warmed to body temperature. The incubator also adjusts the pH of the culture media to the human body. The most commonly used environmental conditions for human IVF incubators are 5% CO2 in air, 37 degrees Centigrade (98.6 F), and 100% relative humidity. Regular air is about 78% nitrogen (N2) and 22% oxygen (O2). The CO2 content of air is less than one tenth of 1%. Some labs culture embryos in a low oxygen system using approximately 5% CO2, 5% O2 and 90% N2. 
         [0009]    The fertilization process takes approximately 10-20 hours. Embryos are then cultured for 3-5 days before being replaced back in the uterus by a simple technique much like intrauterine insemination. 
         [0010]    A variety of coverslips (a thin glass plate) and clamps are often used to cover the sample dishes to contrive individual sealed chambers within the incubator. These devices tend to be complicated, large, and cumbersome to handle and put the sample at risk of “sloshing” around inside and getting lost in the dish. 
         [0011]    The ability to use existing incubators to store multiple chamber devices on each shelf and maintain the integrity of the culture environment and the identification of each sample as separate from each other present challenges to IVF laboratory. Incubators must maintain a very constant and clean environment for embryo culture. A large amount of rinsing media is equilibrated in the incubator. The fluid is used by the physician to wash the oocyte from the ovarian follicle if it is not found in the initial aspirations. 
         [0012]    The simplest chamber product alternative to a cover slipped dish appears to be the Billups-Rothenberg Chamber but only one or maybe two of their chambers will fit into conventional incubators used in the target market (Human IVF). That&#39;s because the Billups-Rothenberg chamber is a multiple sample or tissue culturing system geared for the biotech laboratory that performs massive tissue culturing colonies as compared to IVF; it is large with shelves inside designed for long-term culturing and/or gas and nutrient addition during the culturing period. 
         [0013]    Incubators are typically about the size of a small refrigerator and are often stacked to save lab space. Colored tape on the door is used to identify the location of a patients eggs or embryos in the incubator before the door is opened. This reduces the amount of time that the door has to remain open. 
         [0014]    Typical use is to remove the culture dish from the incubator to check culture growth every 24 hours under a conventional stereo or inverted microscope and as quickly as possible review the cell&#39;s growth and return the dish to the stable culture environment of the incubator. 
         [0015]    Current practice in IVF is the use of incubators that deliver either single gas concentrations (CO2) or a mixed-gas of O2 and CO2. Different culture media require different gas levels to maintain optimal pH. Conventional incubators struggle to achieve these gas settings after every door opening. For a variety of conventional incubators return to optimal gas levels can take from 5 to as long as 45 minutes with the resultant fluctuations in pH values. This large box recovery requires major financial investment in gas supply, storage, and delivery systems. 
         [0016]    There is need for a compact portable hermetic sealed environmental chamber designed for the IVF lab requirement to maintain one sample per device and check culture growth every 24 hours under a conventional stereo or inverted microscope where typical use would be to remove the sealed chamber with culture dish from the incubator, remove lid, remove culture dish and examine the cell&#39;s growth. The reverse process post examination would then be performed and the lid resealed the gas inside the chamber recharged, and the chamber returned as quickly as possible to the conventional large incubator. 
       SUMMARY OF THE INVENTION 
       [0017]    It is a primary object of the present invention to overcome all the aforementioned challenges by providing a micro chamber to continuously maintain a gaseous environment of high humidity and controlled temperature around single or multiple IVF culture dishes that will support culture growth while situated, for example, within an incubator or on a microscope stage. 
         [0018]    Another object of the invention is the provision of a thermal conductive base to hold the dish securely in place and through radiant heat absorption (device sitting in incubator or on heated plate) provide for temperature stabilization of the cell environment inside the dish. 
         [0019]    Another object of the invention is the provision of an integrated electrical circuit thermal source to maintain a user selected temperature. 
         [0020]    Another object of the invention is the provision of a transparent dome lid for visual inspection of the contents. 
         [0021]    Another object of the invention is the provision of a gasket and latch mechanism to allow a pressured unique and custom gas mixture to be introduced and maintained securely inside the chamber for a minimum of 72 hours and hermetically ensure protection against ambient gas entering the chamber. 
         [0022]    Another object of the invention is the provision of an optically clear (glass or plastic) viewing port allowing microscopic visualization of the dish contents. 
         [0023]    Another object of the invention is the provision of a viewing port that will maintain the temperature integrity of the internal culturing environment by utilizing a heat source integrated to the glass or plastic insert that warms the dish and allows the user to set the and maintain the temperature of the glass or plastic insert. 
         [0024]    Another object of the invention is the provision of an integrated temperature display (analog or digital) in the base to demonstrate temperature of the conductive base structure and in turn the approximate temperature of the culture environment. 
         [0025]    Another object of the invention is the provision in the base of an integrated input and output system of fittings, connectors and valves for introducing a gas mixture to the chamber. 
         [0026]    Another object of the invention is the provision pressure gauge connected to the output fitting to display the achieved pressure inside the chamber. 
         [0027]    Another object of the invention is the base will be sized to fit in the palm of the hand of a typical lab technician to ensure ease of movement from storage environment (incubator, refrigerator, etc) to examining microscope or workbench. 
         [0028]    Another object of the invention is incorporation of a bubble level mechanism into the base to display the levelness of the device. 
         [0029]    Another object of the invention is the base will be capable of repeat sterilizations in conventional autoclaves, dishwashers, or other manual and automatic methods of cleaning and sterilization. 
         [0030]    Another object of the invention is the dome and gasket system will be a single use item designed for a single culturing cycle and then discarded. 
         [0031]    Another object of the invention is the base will incorporate a means to allow the user to monitor the internal temperature and gas status using independent probes integrated into the base. 
         [0032]    Additional features of the present invention will become apparent from the following description and appended claims, taken in conjunction with the accompanying drawings. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0033]    Many aspects of the disclosure can be better understood with reference to the following drawings. The components in the drawings are not necessarily to scale. Moreover, in the drawings, like reference numerals designate corresponding parts throughout the several views. 
           [0034]      FIG. 1  is a top perspective view of the chamber construction in assembled sealed relationship of this invention. 
           [0035]      FIG. 2  is a top perspective view of the chamber base of this invention. 
           [0036]      FIG. 3  is a bottom perspective view of the chamber base of this invention. 
           [0037]      FIG. 4  is a top perspective view of the chamber cover of this invention. 
           [0038]      FIG. 5  is a top plan view of the chamber base of this invention. 
           [0039]      FIG. 6  is a bottom plan view of the chamber base of this invention. 
           [0040]      FIG. 7  is a cross-sectional view of the chamber base construction of this invention taken through line  7 - 7  of  FIG. 5 . 
           [0041]      FIG. 8  is a right side view of the chamber base of this invention. 
           [0042]      FIG. 9  is a cross-sectional view of the chamber base construction of this invention taken through line  9 - 9  of  FIG. 5 . 
           [0043]      FIG. 10  is a front view of the chamber base of this invention. 
           [0044]      FIG. 11  is a cross-sectional view of the chamber base construction of this invention taken through line  11 - 11  of  FIG. 6 . 
           [0045]      FIG. 12  is a cross-sectional view of the chamber base construction of this invention taken through line  12 - 12  of  FIG. 6 . 
           [0046]      FIG. 13  is a top plan view of the chamber cover of this invention. 
           [0047]      FIG. 14  is a cross-sectional view of the chamber cover construction of this invention taken through line  14 - 14  of  FIG. 13 . 
           [0048]      FIG. 15  is a right side view of the chamber cover of this invention. 
           [0049]      FIG. 16  is a front view of the chamber cover of this invention. 
           [0050]      FIG. 17  is a side view of the control stem for the input and the output valves of this invention. 
           [0051]      FIG. 18  is a control knob end view of the chamber input and the output valve controls of this invention. 
           [0052]      FIG. 19  is a perspective view of the chamber base latch pivot of this invention. 
           [0053]      FIG. 20  is a perspective view of the chamber base latch thumbscrew of this invention. 
       
    
    
     DETAILED DESCRIPTION 
       [0054]    Referring to  FIGS. 1-20 , there is shown an illustrative embodiment of the invention device, designated generally  10 , as designed for use by the embryologist to maintain an IVF culturing environment independent of ambient disruptions to ensure integrity of a compete culturing cycle. 
         [0055]      FIG. 1  is a top perspective view of IVF culture chamber  10  which comprises octagonal shaped base  30 ; dome cover  60 , seal o-ring  67  in cover groove  66 , cover latch tab  63  retained by latch thumbscrew  80  to a pivot rotatably held in base sleeve  36 , cover hinge tab  62  retained in base slot  50 , output control valve  40 , control valve stem  78 , external output port  42 , input control valve  70 , external input port  72  with input hose fitting  48  threaded therein, and label recesses  32 . 
         [0056]      FIG. 2  is a top perspective view of uncovered octagonal shaped IVF culture chamber base  30  which comprises internal output port  46 , internal input port  76 , culture dish indents  52  and  54  positioned between the internal ports, major surface recess  56  surrounding the indents and internal ports, thumbscrew notch  38 , latch pivot sleeve  36 , cover hinge slot  50 , output valve socket  44 , input valve socket  74 , external output port  42 , thumbscrew notch  38 , latch pivot sleeve  36 , and label recesses  32 . 
         [0057]      FIG. 3  is a bottom perspective view of base  30  showing pedestal  31 , circular grooves  33 , cross grooves  35 , thumbscrew pivot sleeve  36 , control valve set screw  37 , output control valve socket  44 , thumbscrew notch  38 , and cover hinge tab slot  50 . 
         [0058]      FIG. 4  is a top perspective view of dome cover  60  showing cover hinge tab  62 , cover latch tab  63 , thumbscrew retaining notch  68 , and seal o-ring  67  in retaining groove  66 . 
         [0059]      FIG. 5  is a top plan view of base  30  showing internal input port  76 , internal output port  46 , culture dish indents  52  and  54  placed between internal ports  76  and  46 , thumbscrew notch  38 , cover hinge slot  50 , with line  7 - 7  in reference to cross-section  FIG. 7  and line  9 - 9  in reference to cross-section  FIG. 9 . 
         [0060]      FIG. 6  is a bottom plan view of base  30  showing fluid communication between external input port  72  and internal input port  76  through control valve socket  74 , fluid communication between external output port  42  and internal output port  46  through control valve socket  44 , culture dish indents  52  and  54  placed between internal ports  76  and  46 , thumbscrew notch  38 , cover hinge slot  50 , line  11 - 11  reference to cross-section  FIG. 11 , and line  12 - 12  reference to cross-section  FIG. 12 . 
         [0061]      FIG. 7  is a cross-section view of base  30  along line  7 - 7  of  FIG. 5  showing pedestal  31 , circular grooves  33 , culture dish indents  52  and  54 , surrounding major surface recess  56 , and latch pivot sleeve  36 . 
         [0062]      FIG. 8  is a right side view of base  30  showing pedestal  31 , cross groove  35 , latch pivot sleeve  36 , external input port  72 , and control valve socket  74 . 
         [0063]      FIG. 9  is a cross-section view of base  30  along line  9 - 9  of  FIG. 5  showing pedestal  31 , circular grooves  33 , culture dish indents  52  and  54 , internal port  46  with its fluid communication to control valve socket  44 , internal port  76  with its fluid communication to control valve socket  74 , and surrounding major surface recess  56 . 
         [0064]      FIG. 10  is a front view of base  30  showing pedestal  31 , cross groove  35 , latch pivot sleeve  36 , thumbscrew notch  38 , external output port  42 , major surface recess  56 , and external input port  72 . 
         [0065]      FIG. 11  is a cross-section view of base  30  along line  11 - 11  of  FIG. 6  showing external input port  72  with its fluid communication to control valve socket  74 , latch pivot sleeve  36 , major surface recess  56 , label recess  32 , pedestal  31 , and circular groove  33 , cross groove  35 . 
         [0066]      FIG. 12  is a cross-section view of base  30  along line  12 - 12  of  FIG. 6  showing pedestal  31 , circular groove  33 , cross groove  35 , latch pivot sleeve  36 , thumbscrew notch  38 , and major surface recess  56 . 
         [0067]      FIG. 13  is a top plan view of dome cover  60  showing cover latch tab  63 , thumbscrew retaining notch  68 , cover hinge tab  62 , circumferential seal  67  in retaining groove  66 , and line  14 - 14  reference to cross-section  FIG. 14 . 
         [0068]      FIG. 14  is a cross-section view of dome cover  60  along line  14 - 14  of  FIG. 13  showing cover hinge tab  62 , cover latch tab  63 , and seal o-ring  67  in retaining groove  66 . 
         [0069]      FIG. 15  is a right side view of dome cover  60  showing cover latch tab  63 , cover hinge tab  62 , and seal o-ring  67 . 
         [0070]      FIG. 16  is a front view of dome cover  60  showing cover latch tab  63 , cover hinge tab  62 , and seal o-ring  67 . 
         [0071]      FIG. 17  is a side view of control valve stem  78  which is retained by sockets  44  and  74  for valves  40  and  70  respectively showing external port seal o-ring groove  43 , internal port seal o-ring groove  47 , stem thread surface  41 , and valve control knurled knob  45 . 
         [0072]      FIG. 18  is a knob-end view of control valve stem  78 . 
         [0073]      FIG. 19  is a perspective view of latch pivot  86  which is positioned rotatively in sleeve  36  of base  30  and secured longitudinally by thumbscrew  80  threaded into latch pivot hole  88  via notch  38  in base  30 . 
         [0074]      FIG. 20  is a perspective view of thumbscrew  80  having threads  82  which screw into latch pivot hole  88  to secure dome cover  60  to base  30  compressively through gasket  67  by advancing thumbscrew flange  84  against cover latch tab  63  to close hermetically chamber  10 . 
         [0075]    It is a primary object of the present invention to provide a micro chamber  10  to continuously maintain a gaseous environment of high humidity and controlled temperature around single or multiple IVF culture dishes that will support culture growth while situated, for example, within an incubator or on a microscope stage. 
         [0076]    Micro chamber  10  is sized to fit in the palm of the hand of a typical lab technician to ensure ease of movement from an incubator to an examining microscope or workbench for use in conjunction with one or more culture dishes. 
         [0077]    Base  30  is thermally conductive and has indents  52  and  54  to hold the culture dish securely in place to provide for temperature stabilization of the cell environment inside the dish through radiant heat absorption when the chamber  10  is sitting in an incubator or on a heated plate, the indents  52  and  54  being matched to the feet or underside of the culture dish so the underside of the dish is in full contact with base  30  thereby improving conductive temperature transfer between base  30  and the dish. 
         [0078]    Base  30  bottom has a concentric pattern of circular and radial grooves capable of preventing thermal warp while insuring heat radiation. 
         [0079]    Base  30  is capable of repeat sterilizations in conventional autoclaves, dishwashers, or other manual and automatic methods of cleaning and sterilization. 
         [0080]    Domed cover  60  is made of see-through material for visual inspection of the chamber contents, cover  60  being a single use item designed for a single culturing cycle and then discarded. 
         [0081]    Gasket  67  retained in cover groove  66  and latch mechanism tab  63  retained by thumbscrew  80  and hinge cover hinge tab  62  retained in base slot  50  allow a pressured unique and custom gas mixture to be introduced and maintained securely inside the chamber for a minimum of 72 hours and ensure protection against ambient gas entering the chamber, the gasket  67  being an O-ring of the gland type retained in cover groove  66  to provide an hermetic seal between the opposed base  30  and cover  60  which are designed as a unit for pressurization, gasket  67  being a single use item designed for a single culturing cycle and then discarded. 
         [0082]    Gas input and output systems for purge, fill, and pressurization of sealed chamber  10  are integrated in base  30  for one-hand operation by input control valve  70  in socket  74  communicating with external input port  72  and internal input port  76  and by output control valve  40  in socket  44  communicating with internal output port  46  and external output port  42 . 
         [0083]    An error free identification system linking micro chamber  10  by text/bar code label to its culture dish is provided for by recesses  32  in base  30  outside domed cover  60 . 
         [0084]    Base top major surface recess  56  surrounds culture dish indents  52  and  54  to confine and conduct spillage to the front and rear sides of base  60  for disposal.