Abstract:
Exemplary embodiments of apparatus and method for facilitating an analysis of a sample(s) can be provided. For example, using a first arrangement(s), it can be possible to receive the sample(s) thereon. Further, for example, using a second arrangement(s), it can be possible to cause a force to be applied on a portion(s) of the sample(s) such that a surface(s) of the sample(s) can be flattened against a section(s) of the first arrangement(s).

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This present application relates to and claims priority from International Patent Application No. PCT/US2013/054653 filed Aug. 13, 2013, and from U.S. Patent Application No. 61/682,407, filed on Aug. 13, 2012, the entire disclosures of which are incorporated herein by reference. 
    
    
     STATEMENT OF FEDERAL SUPPORT 
     The present disclosure was made with U.S. Government support under grant numbers R01EB002715 and R01 EB012466 from the National Institute of Health. Thus, the Government has certain rights to the disclosure described and claimed herein. 
    
    
     FIELD OF THE DISCLOSURE 
     Exemplary embodiments of the present disclosure relate to imaging and application of microscopy to anatomical structures, and more particular to devices applicable to any type of tissue(s) which can facilitate confocal microscopy, optical microscopy and/or imaging. 
     BACKGROUND INFORMATION 
     A complete and accurate surgical excision of cancers, while preserving as much of the surrounding normal tissue as intact as possible, can be guided by the examination of pathology for residual cancer margins. However, a preparation of the pathology can be labor-intensive, and can be time-consuming. The processing of excised tissue and preparation of thin sections for pathology can take hours (e.g., for frozen sections) or days (e.g., for fixed sections), which can result in insufficient sampling of tissue, and inaccurate and/or incomplete removal of cancer. Consequently, a large number (e.g., between about 20% and 70%, depending on the setting) of patients undergo re-excision (e.g., repeat surgery) and/or chemotherapy and/or radiotherapy. 
     Confocal microscopy can image nuclear and cellular morphology in living tissues, either in vivo or in freshly excised, or biopsied, tissue ex vivo, without the need for processing tissue or preparation of thin sections. A detection of residual cancer margins can be made possible in fresh tissue within minutes. Rapid mosaicking, for example, acquisition and stitching together of a large number of images, can facilitate imaging over large areas. 
     Accordingly, there may be a need to address and/or overcome at least some of the above-described deficiencies and limitations, and to provide exemplary embodiments of devices according to the present disclosure as described in further details herein. 
     SUMMARY OF EXEMPLARY EMBODIMENTS OF THE DISCLOSURE 
     Indeed, one of the objects of certain exemplary embodiments of the present disclosure can be to address the exemplary problems described herein above, and/or to overcome the exemplary deficiencies commonly associated with the prior art as, for example, described herein. Accordingly, for example, provided and described herein are certain exemplary embodiments of exemplary devices according to the present disclosure which can be applicable to tissue(s) which facilitates confocal microscopy, optical microscopy and/or imaging. 
     Due to the three-dimensional (“3D”) topography and irregular shapes and sizes of fresh surgically excised, or biopsied, tissue, mounting the tissue for imaging large areas with a scanning confocal microscope, or other modalities, as mentioned above, can be challenging due to the following problems:
         a. Sag, for example, bending of the desired tissue surface (e.g., imaging plane) to be imaged.   b. Tissue stability during imaging and mosaicing procedures.   c. Uniform tissue flatness for the surface to be imaged.   d. Constant hydration of tissue since it is fresh and living.   e. Orthogonality of the desired tissue surface (e.g., imaging plane) relative to optic axis of objective, for example, desired tissue surface to be imaged must be parallel to the “object plane” of the microscope.       

     Confocal mosaicing microscopy, and possibly other emerging/competing optical imaging modalities, such as optical coherence tomography, multiphoton microscopy, etc. can facilitate technologies for rapid pathology at the bedside in large amounts of fresh tissue. One of the important factors to image a large area of the fresh tissue can be that the entire imaging surface should be parallel to the imaging plane. 
     According to an exemplary embodiment of the present disclosure, it can be preferable to provide a technology platform, which can be called “confocal mosaicing microscopy,” to facilitate a rapid pathology at the bedside. According to such exemplary embodiment, it can be beneficial to mount surgically-excised tissue in a microscope. A device according to a certain exemplary embodiment of the present disclosure can be provided for mounting fresh tissue from surgery. While the exemplary embodiment described herein can be directed to the use of a fresh tissue from surgical excisions for use in surgical settings, such exemplary device can also be used for biopsies in clinical settings. 
     For example, with respect to a diverse range of tissues (e.g., skin, breast, head-and-neck or otolaryngologic, urologic, brain or neurologic, etc.) and wide range of settings sizes and shapes (e.g., large excisions, thin excisions, shave biopsies, punch biopsies, needle core biopsies, fine needle aspirations, etc.), it can be possible to utilize and/or apply exemplary embodiments of such device to various targets and/or tissues. Further, the exemplary embodiments of the device, according to the present disclosure, can also be used with other present and future optical imaging modalities, such as optical coherence tomography, reflectography, scanning electrochemical microscopy, multiphoton microscopy, etc. 
     These and other objects of the present disclosure can be achieved by provision of an exemplary apparatus for facilitating an analysis a sample(s), which can include a first arrangement(s) which can be configured to receive the sample(s) thereon, and a second arrangement which can be configured to apply a force on a portion(s) of the sample(s) such that a surface(s) of the sample(s) can be flattened against a section(s) of the first arrangement(s). The second arrangement(s) can be configured to apply the force to an area of the portion(s) that can be located on a further surface that can be approximately opposite to the surface(s). The second arrangement(s) can include an inflatable arrangement(s), a piston(s), a cassette lid(s), and/or a plurality of pins. The second arrangement(s) can also include a flexible tissue holding arrangement(s) and a vacuum arrangement(s). The flexible tissue holding arrangement(s) can include a silicon bag(s). 
     In some exemplary embodiments of the present disclosure, a third arrangement can be configured to directly or indirectly secure the sample(s) in a position to maintain the surface(s) in a flat manner against the section(s). A fourth arrangement(s) can be configured to obtain data regarding a portion(s) of the sample(s) from below the flattened surface(s). The fourth arrangement can include a microscope arrangement, and the data can include image information regarding the portion(s) of the sample(s). 
     In another embodiment of the present disclosure can be a method for facilitating an analysis of a sample(s), which can include providing an arrangement(s) so as to receive the sample(s) thereon, providing the sample(s) on the arrangement(s), and causing a force to be applied on a portion(s) of the sample(s) such that a surface(s) of the sample(s) can be flattened against a section(s) of the arrangement(s). The force can be applied using a further arrangement(s), which can include an inflatable arrangement(s). The further arrangement(s) can include an inflatable arrangement holder(s) and a piston(s). The inflatable arrangement(s) can be inflated, and the piston(s) can be screwed into the inflatable arrangement holder(s) to cause the inflatable arrangement(s) to apply the force to the sample(s). In some exemplary embodiments of the present disclosure, the further arrangement can include a cassette lid(s). The inflatable arrangement(s) can be inflated, and the cassette lid(s) can be closed to cause the inflatable arrangement(s) to apply the force to the sample(s). The further arrangement(s) can include a plurality of pins. 
     In certain exemplary embodiments of the present disclosure, the further arrangement(s) can includes a flexible tissue holding arrangement(s) and a vacuum arrangement(s). The sample(s) can be placed in the flexible tissue holding arrangement(s), and the air can be vacuumed from the flexible tissue holding arrangement(s) using the vacuum(s) to apply the flexible tissue holding arrangement(s). The flexible tissue holding arrangement(s) can include a silicon bag(s). 
     These and other objects, features and advantages of the exemplary embodiment of the present disclosure will become apparent upon reading the following detailed description of the exemplary embodiments of the present disclosure, when taken in conjunction with the appended claims. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       For a more complete understanding of the present disclosure and its advantages, reference is now made to the following description, taken in conjunction with the accompanying drawings, in which: 
         FIG. 1  is a side cross-sectional view of an exemplary tissue-mounting device for fresh tissue, with no force applied to a bladder, in accordance with a first exemplary embodiment of the present disclosure; 
         FIG. 2  is a side cross-sectional view of the exemplary tissue-mounting device of  FIG. 1  with an inserted piston, during operation thereof according to an exemplary embodiment of the present disclosure; 
         FIG. 3  is a side cross-sectional view of the exemplary tissue-mounting device of  FIG. 1  installed in an exemplary microscope according to an exemplary embodiment of the present disclosure; 
         FIG. 4  is a perspective isometric (e.g., 3D) view of the exemplary tissue-mounting device of  FIG. 1 , with separate exemplary components illustrated therein according to an exemplary embodiment of the present disclosure; 
         FIG. 5  is a side cross-sectional view of another exemplary embodiment of the exemplary tissue-mounting device for fresh tissue, with no force applied to the bladder according to a second exemplary embodiment of the present disclosure; 
         FIG. 6  is a side cross-sectional view of the exemplary tissue-mounting device of  FIG. 5 , with the exemplary cassette closed, during operation thereof according to an exemplary embodiment of the present disclosure; 
         FIG. 7  is a side cross-sectional view of the exemplary tissue-mounting device of  FIG. 5  installed in an exemplary microscope according to an exemplary embodiment of the present disclosure; 
         FIG. 8  is a perspective isometric (e.g., 3D) view of the exemplary tissue-mounting device of  FIG. 5 , with separate exemplary components illustrated therein according to an exemplary embodiment of the present disclosure; 
         FIG. 9  is another perspective isometric (e.g., 3D) view of the exemplary tissue-mounting device of  FIG. 5 , with further separate exemplary components illustrated therein according to an exemplary embodiment of the present disclosure; 
         FIG. 10  is a side cross-sectional view of another exemplary tissue-mounting device for fresh tissue using a plurality of exemplary pins according to a third exemplary embodiment of the present disclosure; 
         FIG. 11  is a side cross-sectional view of still another tissue-mounting device for fresh tissue using an exemplary silicon bag and an exemplary vacuum according to a fourth exemplary embodiment of the present disclosure; 
         FIG. 12A  is a side view of a possible result when the tissue is not flattened according to an exemplary embodiment of the present disclosure; 
         FIG. 12B  is a side view of a possible result when the tissue is flattened using one or more exemplary devices according to the present disclosure according to an exemplary embodiment of the present disclosure; 
         FIGS. 13A-16  are exemplary images of an exemplary embodiments of the exemplary tissue-mounting device in various stages of an exemplary operation according to an exemplary embodiment of the present disclosure; 
         FIG. 17  is an exemplary flow diagram for flattening the exemplary tissue according to an exemplary embodiment of the present disclosure; 
         FIG. 18  is an exemplary flow diagram for flattening the exemplary tissue according to another exemplary embodiment of the present disclosure; 
         FIG. 19  is an exemplary flow diagram for flattening the exemplary tissue according to the third exemplary embodiment of the present disclosure; and 
         FIG. 20  is an exemplary flow diagram for flattening the exemplary tissue according to the fourth exemplary embodiment of the present disclosure. 
     
    
    
     Throughout the figures, the same reference numerals and characters, unless otherwise stated, are used to denote like features, elements, components or portions of the illustrated embodiments. Moreover, while the subject disclosure will now be described in detail with reference to the figures, it is done so in connection with the illustrative embodiments and is not limited by the particular embodiments illustrated in the figures and the accompanying claims. It is intended that changes and modifications can be made to the described embodiments without departing from the true scope and spirit of the subject disclosure and the accompanying claims. 
     DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS 
     In order to image a large area of the tissue with nuclear and cellular level resolution, the tissue can be scanned in a two-dimensional (“2D”) “object plane” (e.g., a plane that can be approximately perpendicular to an optical axis of an objective lens) in the microscope. However, surgically excised, and biopsied, tissue can have a 3D topography with varying shape and size. Thus, it can be preferable that the tissue, especially the surface to be imaged, be flattened into a 2D plane that can conform to the object plane of the microscope, as shown in  FIGS. 12A and 12B . 
     To that end, an exemplary embodiment of the tissue-mounting device according to the present disclosure can be provided to perform such exemplary procedure, as shown in  FIGS. 1-4 . 
     In particular,  FIG. 1  illustrates a side cross-sectional view of an exemplary embodiment of the tissue-mounting device  100 , with no force applied to a bladder  125 , which can perform an exemplary procedure shown in  FIG. 17 . To flatten the desired (e.g., lower) surface of the tissue  105 , a user can place the tissue  105  at the center of a window  110  of such exemplary tissue-mounting device  100  (e.g., procedure  1705  in  FIG. 17 ). Then, the user can apply a surgical lubricant on the top surface of the window  110  (e.g., the surface where the tissue can be placed) around the sample, for example, approximately 5 mm outside a border of the sample (e.g., procedure  1710  in  FIG. 17 ). Then, the window  110  can be placed on a tip-tilt plate  115  (e.g., procedure  1715  in  FIG. 17 ) followed by installation of a polycarbonate bladder-chamber  120  on the tip-tilt plate  115  (e.g., procedure  1720  in  FIG. 17 ). The exemplary tissue-mounting device  100  for fresh tissue  105  shown in  FIG. 1  can have no force applied to the bladder  125 . Under this nominal condition, the tissue surface (e.g., a lower surface of tissue  105 ), which can be imaged, may not be flattened on to the object plane. 
       FIG. 2  shows a side cross-sectional view of the exemplary tissue-mounting device  100  of  FIG. 1  with an inserted piston  205 , during operation thereof. In particular, as shown in  FIG. 4 , which illustrates a perspective isometric (e.g., 3D) view of the exemplary tissue-mounting device  100  of  FIG. 1 , with separate exemplary components illustrated therein, a polycarbonate piston  205  can be inserted into the bladder-chamber  120  of the tissue-mounting device  100  of  FIG. 1  (e.g., procedure  1725  in  FIG. 17 ), and pressed down gently to apply pressure on the bladder  125  (e.g., procedure  1730  in  FIG. 17 ). This operation and force can flatten a lower surface of the tissue  105  against the window  110 , and thus provide the tissue  105  into a desired 2D plane. The exemplary system/apparatus, which can be used to facilitate such flattening of the tissue, is illustrated in  FIG. 4 , which can facilitate the use of a microscope to analyze the flattened surface of the tissue. 
     According to an exemplary embodiment of the present disclosure, position and orientation on a 2D plane in terms of tip and tilt, relative to the optical axis of the microscope, can be provided as shown in  FIG. 3 , which illustrates a side cross-sectional view of the exemplary tissue-mounting device  100  of  FIG. 1  installed in a microscope  300 , which can be adjusted with spring-loaded thumbscrews  405  shown in  FIG. 4 . This can facilitate an exemplary adjustment of the window  110  so that it can be parallel to the “object plane” of the microscope. Such exemplary system and method, according to an exemplary embodiment of the present disclosure can facilitate a desired surface of the tissue  105  to conform to the object plane of the microscope  300 , to facilitate accurate and repeatable imaging and mosaicing over large areas. The exemplary embodiment and implementation of the mounting device can be robust to facilitate a repeatable operation during extended periods of time. Thus, the use of such fresh tissue mounting devices in confocal mosaicing microscopy, can enable rapid pathology at the bedside in diverse settings (e.g., for detection of residual cancer margins to guide surgery in surgical settings, and for screening/diagnosis of cancers to guide the examination of biopsies in clinical settings). 
     The exemplary flattening procedure illustrated in  FIG. 18  can also be accomplished with another mounting device according to a further exemplary embodiment of the present disclosure, as illustrated in  FIGS. 5-9 . Thus, for example, as an initial step for the exemplary procedure to flatten the desired (e.g., lower) surface of the tissue  505 , the user can place the tissue  505  at a center of the cassette  510  (e.g., procedure  1805  of  FIG. 18 ), followed by placing the bladder  515  on the sample (e.g., procedure  1810  of  FIG. 18 ), which is shown in  FIG. 5 . Such placement can also be done automatically by an automatic device, such as, for example, a robotic device, etc. Then, the user and/or the automatic device can close the cassette lid  520  (e.g., procedure  1815  of  FIG. 18 ). Such exemplary closure of the lid  520  can apply pressure on the bladder  515  which can flatten the lower surface of the tissue  505  against the bottom surface of the cassette  510  into the desired 2D plane, as shown in  FIG. 6 . Then, as also illustrated in  FIG. 6 , the cassette  510  can be placed on the tip-tilt plate  525 , and locked down with the holding clamps  605  (e.g., procedure  1020  of  FIG. 18 ). 
     The 2D plane&#39;s position and orientation in terms of tip and tilt, relative to the optical axis of the microscope, can be adjusted with spring-loaded thumbscrews  530 . This can facilitate an adjustment of the window so that it can be parallel to the object plane or the image plane  705 , or approximately perpendicular to the optical axis of the microscope  710 , as shown in  FIG. 7 . 
       FIG. 8  is a perspective isometric (e.g., 3D) view of the exemplary tissue-mounting device of  FIG. 5 , with separate exemplary components illustrated therein.  FIG. 9  provides another perspective isometric (e.g., 3D) view of the exemplary tissue-mounting device of  FIG. 5 , with such separate exemplary components illustrated therein. 
     The exemplary flattening procedure illustrated in  FIG. 19  can also be accomplished with another mounting device according to a further exemplary embodiment of the present disclosure, as illustrated in  FIG. 10 . To flatten the desired (e.g., lower) surface, the user can place the fresh tissue  1005  on the glass imaging window  1010  (e.g., procedure  1905  of  FIG. 19 ) and then placing the micro-pin array  1015  on the tissue  1005  (e.g., procedure  1910  of  FIG. 19 ). Then, the user can apply a force on the pin array  1015  to flatten the lower surface of the tissue  1005  against the top surface of the imaging window  1010  into the desired 2D plane (e.g., procedure  1915  of  FIG. 19 ). An imaging device  1030 , which can include an objective lens  1020  and a laser beam  1025 , can be used to image to fresh tissue  1005  (e.g., procedure  1920  of  FIG. 19 ). 
     The exemplary flattening procedure illustrated in  FIG. 20  can also be accomplished with another mounting device according to a further exemplary embodiment of the present disclosure, as illustrated in  FIG. 11 . To flatten the desired (e.g., lower) surface the user can place the fresh tissue  1105  on the imaging window  1110  followed by placing the tissue/glass window in a bag (e.g., a thin silicon bag)  1115  (e.g., procedure  2005  of  FIG. 20 ). Then, the user can apply a vacuum  1120  to the silicon bag  1115  (e.g., procedure  2010  of  FIG. 20 ). This can apply atmospheric pressure, flattening the tissue  1105  against the top surface of the imaging window  110  into the desired 2D plane (e.g., procedure  2015  of  FIG. 20 ). 
       FIG. 12A  shows a side view of an exemplary illustration of a result of the tissue that may not be flattened, and thus, the object plane  1205  can likely not be parallel to the image plane  1210 , and the optical axis may not be orthogonal to the image plane.  FIG. 12B  provides a side view of a possible result when the tissue can be flattened using one or more exemplary devices according to the present disclosure, thus making the object plane parallel to the image plane and the optical axis orthogonal to the image plane. 
     Tissue fixturing can be utilized when acquiring large number of images for mosaicing. Indeed, the exemplary desired tissue surface (e.g., to be imaged) can preferably be flattened, and positioned and oriented so as to be held approximately parallel to the microscope objective lens&#39; object (e.g., focal) plane. Thus, when the tissue can be translated in, for example, two dimensions, the surface can remain at least approximately in the lens&#39; focal plane. If the tissue surface can be tilted, then the lens&#39; focal plane (e.g., imaging) can either “sink into” or “lift off” the tissue surface. 
       FIGS. 13A-16  show various illustrations of exemplary embodiments of the exemplary tissue-mounting devices in various stages of an exemplary operation. For example,  FIG. 13A  illustrates the exemplary tissue mounting device before an exemplary sample is placed thereon.  FIG. 13B  illustrates the exemplary tissue mounting device having an exemplary slide/window  1305  placed thereon.  FIG. 13C  illustrates the exemplary tissue mounting device having an exemplary plate  1310  placed on the exemplary slide/window  1305 .  FIG. 13D  has an exemplary bladder chamber  1315  placed on the exemplary plate  1310 .  FIG. 13E  illustrates the exemplary tissue mounting device having an exemplary piston holder  1320  placed on the exemplary bladder holder  1315 .  FIGS. 13F and 14  illustrate the exemplary tissue mounting device having an exemplary tissue sample  1325  placed inside of the exemplary bladder chamber  1315 .  FIGS. 15A-15C  illustrate the exemplary tissue mounting device having an exemplary bladder  1305  placed inside of the exemplary bladder chamber  1315 .  FIG. 15D  illustrates the exemplary tissue mounting device having a ball bearing ring  1510  placed inside of the bladder chamber  1315 .  FIG. 15E  illustrates the exemplary tissue mounting device having a cover  1515  placed thereon.  FIG. 16  illustrates the exemplary tissue mounting device being used to image an exemplary tissue sample  1325 . 
     The foregoing merely illustrates the principles of the disclosure. Various modifications and alterations to the described embodiments will be apparent to those skilled in the art in view or the teachings herein. It will thus be appreciated that those skilled in the art will be able to devise numerous systems, arrangements, and procedures which, although not explicitly shown or described herein, embody the principles of the disclosure and can be thus within the spirit and scope of the disclosure. In addition, all publications and references referred to above can be incorporated herein by reference in their entireties. It should be understood that the exemplary procedures described herein can be stored on any computer accessible medium, including a hard drive, RAM, ROM, removable disks, CD-ROM, memory sticks, etc., and executed by a processing arrangement and/or computing arrangement which can be and/or include a hardware processors, microprocessor, mini, macro, mainframe, etc., including a plurality and/or combination thereof. In addition, certain terms used in the present disclosure, including the specification, drawings and claims thereof, can be used synonymously in certain instances, including, but not limited to, for example, data and information. It should be understood that, while these words, and/or other words that can be synonymous to one another, can be used synonymously herein, that there can be instances when such words can be intended to not be used synonymously. Further, to the extent that the prior art knowledge has not been explicitly incorporated by reference herein above, it can be explicitly being incorporated herein in its entirety. All publications referenced can be incorporated herein by reference in their entireties.