Abstract:
A method for detecting cancer includes providing a biomedical sample, performing a first adsorption step, performing a first desorption step, and performing a first discrimination step. The first adsorbing step includes immersing the biomedical sample into a first detection reagent having a first adsorbent that adsorbs onto the biomedical sample. The first desorption step includes immersing the biomedical sample into a first desorbing agent for a first period of time. The first discrimination step includes measuring an amount residual of first adsorbent adsorbed on the biomedical sample for identifying distribution of cancer cells within the biomedical sample. The present invention could be widely applied for detecting various cancers based on the differential physisorption of adsorbent. The present invention can provide a method for rapidly and non-invasively detecting cancers.

Description:
FIELD OF THE INVENTION 
       [0001]    The present invention relates to a method for detecting cancer, and more particularly to a method for detecting cancer based on physisorption kinetics of reagent. 
       DESCRIPTION OF THE PRIOR ART 
       [0002]    At an early phase of cancer, it is difficult to diagnose cancer in a clinic as it is usually asymptomatic. Generally speaking, there is apparently symptomatic for a patient with cancer in the intermediate or advanced phase, which may seek medical assistance for diagnosing and cancer therapy, but by then cancer cells would have metastasized to other organs. Therefore, it is quite important for researcher to develop an easy and rapid method for detecting cancers of early phase. 
         [0003]    Many tumor biomarkers, e.g. CEA (carcinoembryonic antigen) and EGTM (European Group on Tumour Marker), have been developed for various cancers in the past. However, poor issues of sensitivity and specificity for these biomarkers and time-consuming procedures have made these techniques unattractive. Therefore, the tumor markers are presently being applied for tracing relapse of cancers after cancer therapy rather than for the purpose of early diagnosis. 
         [0004]    U.S. Pat. No. 5,726,061 discloses applications of biochemical enzymes for detecting disaccharide markers (-D-Gal(1-3)-D-GalNAc(1-Thr/Ser) and monosaccharide markers (2-acetamido-2-deoxy-D-galactose). However, specific antibodies production is a requisite for detecting various tumors antigen and therefore, the said method is limited. 
         [0005]    U.S. Pat. No. 6,187,591 discloses using a Schiff&#39;s reagent to react with the colon mucosa for detecting the presence of the biomarker, a C12-C20 aliphatic aldehyde, of the colorectal cancer tissue, for early diagnosis for colorectal cancer. However, this diagnostic method is difficultly applied to cell level of detection. 
         [0006]    In addition, genetic testing may be performed for detecting oral cavity cancer; however, this technique is complicated and time-consuming. 
         [0007]    Therefore, it is highly desirable to develop a simple and rapid diagnostic technique for diagnosing various cancers at early phase. 
       SUMMARY OF THE INVENTION 
       [0008]    The present invention is directed to provide a method for diagnosing cancer at early phase so that the cancer may be timely detected and treated. 
         [0009]    The present invention is directed to a method based on physisorption kinetics of reagent for rapidly and non-invasively detecting cancers at early phase. 
         [0010]    The present invention is also directed to reagents applied in the method for diagnosing cancers. According to an embodiment, the physisorption kinetics of the reagent for cancer tissue is dramatically different compared to that of the normal tissue, and therefore it is easily, rapidly and widely applied to detect various types of cancers. 
         [0011]    In one embodiment, a method for detecting cancer includes providing a biomedical sample, performing a first adsorption step, performing a first desorption step and performing a first discrimination step. In the first adsorption step, the biomedical sample is immersed into a first detection reagent including a first adsorbent, which physically adsorbs onto the biomedical sample. In the first desorption step, the biomedical sample is immersed into a first desorption reagent and allowed to react for a first period of time. In the first discrimination step, the amount of residual of first adsorbent adsorbing on the biomedical sample is detected to determine the distribution of cancer cells within the biomedical sample. 
         [0012]    According to another embodiment, a detection reagent comprising an adsorbent is provided for detecting cancer. A method for detecting cancer using the detection reagent includes providing a biomedical sample, performing a first reagent-adsorption step, performing a first reagent-desorption step and performing a first discrimination step. In the first reagent-adsorption step, the biomedical sample is immersed into a first screening reagent including a first adsorbent that physically adsorbs onto the biomedical sample. In the first reagent-desorption step, the biomedical sample is immersed into a first desorption reagent and allowed to be desorbed for a predetermined period of time. In the first discrimination step, the amount of residual of first adsorbent adsorbed on the biomedical sample is measured after being desorbed with a first desorption reagent for determining the distribution of cancer cells within the biomedical sample. 
         [0013]    The objective, technologies, features and advantages of the present invention will become concretely from the following description in conjunction with the accompanying drawings wherein are proposed, by way of illustration and taking example, certain embodiments of the present invention. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0014]    The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. 
           [0015]    The above aspects and advantages of this invention will become to be more readily appreciated, that is, becomes easily to be understood by reference to the following detailed description aided with the accompanying drawings, wherein: 
           [0016]      FIG. 1  is a flow chart illustrating a method for detecting cancer according to an embodiment of the present invention; 
           [0017]      FIG. 2  is a flow chart illustrating a method for detecting cancer according to an embodiment of the present invention; 
           [0018]      FIG. 3  illustrates infrared spectral images illustrating adsorption of paraffin on a cell sample; 
           [0019]      FIG. 4  illustrates infrared spectral images illustrating adsorption of beeswax on a cell sample; 
           [0020]      FIG. 5  illustrates infrared spectral images illustrating adsorption of paraffin on a cell sample; 
           [0021]      FIG. 6  illustrates infrared spectral images illustrating adsorption of beeswax on a cell sample; 
           [0022]      FIG. 7  illustrates infrared spectral images illustrating adsorption of triacontane (C 30 H 62 ) on a cell sample; 
           [0023]      FIG. 8  illustrates infrared spectral images illustrating adsorption of ethyl myristate (C 16 H 32 O 2 ) on a cell sample; 
           [0024]      FIG. 9  illustrates infrared spectral images illustrating adsorption of 3,5,5-Trimethylhexyl 3,5,5-trimethylcaproat (C 20 H 40 O 2 ) on a cell sample; 
           [0025]      FIG. 10  illustrates infrared spectral images illustrating adsorption of paraffin on a tissue sample; 
           [0026]      FIG. 11  illustrates infrared spectral images illustrating adsorption of beeswax on a tissue sample; and 
           [0027]      FIG. 12  illustrates fluorescence images illustrating adsorption of dye-wax reagent on a tissue sample. 
       
    
    
     DESCRIPTION OF THE PREFERRED EMBODIMENT 
       [0028]    The composition of membrane of a cancer cell is different from that of a normal cell, and thus physisorption capability of the cancer cell is different from that of a normal cell. The present invention employs the physisorption capability for differentiating a cancer cell from a normal cell. 
         [0029]    The present invention provides a method for detecting cancer based on above principle. Referring to  FIG. 1 , a flow chart illustrates a method for detecting cancer, which may be described as follows. At step S 11 , a biomedical sample including, such as a tissue section or cell lines, is provided. 
         [0030]    At first reagent adsorption step S 12 , the biomedical sample is immersed into a first detection reagent comprising a first adsorbent, which is physically adsorbed onto the biomedical sample. The physisorption of the first adsorbent is greater for the cancer cells than that of the normal cells in the biomedical sample. For example, the first adsorbent comprises a long-chain ester containing 16-46 carbon atoms. 
         [0031]    Alternatively, the physisorption capability of the first adsorbent is greater for the normal cells than that of the cancer cells in the biomedical sample. For example, the first adsorbent comprises a long-chain alkane containing 21-30 carbon atoms. 
         [0032]    Next, at first reagent-desorption step S 13 , the biomedical sample is immersed into a first desorption reagent for a predetermined period of time. For example, the first desorption reagent comprises an organic solvent for desorbing the first adsorbent adsorbed onto the biomedical sample. The cells with comparatively less physisorption capability for the first adsorbent is more likely to be desorbed with the first desorption reagent, and vice versus. 
         [0033]    The length of the predetermined time depends on many factors, such as the option and concentration of the first adsorbent and the first desorbing reagent. Additionally, according to differential physisorption capability of cancer cells at different cancer stages. Accordingly, the predetermined time can be adjusted according to the amount of residual of adsorbed first adsorbent, which is a signpost for cancer being at various stages. Therefore, based on the relative amount of residual of adsorbent, the cancer phases of the cancer samples may be rapidly determined. 
         [0034]    At discrimination step S 14 , the amount of residual of the first adsorbent adsorbed on the biomedical sample is determined for the distribution of cancer cells within the biomedical sample. In one embodiment, the amount residual of the first adsorbent adsorbed on the biomedical sample is determined by the absorbance or intensity based on the integration in the spectral range of 3000-2800 cm −1  for infrared spectrum or Raman scattering spectrum. 
         [0035]    In another embodiment, the first detection reagent is mixed with a specific fluorescent dye that is soluble in organic solvent and does not bind to lipid molecules or membrane of cell lines or tissue samples. Preferably, the fluorescent dye comprises LC6500, 4-dicyanomethylene-2-methyl-6-p-dimethylaminostyryl-4H-pyran (DCM), Allophycocyanin (APC) or APC-Cy7, for example. The amount of residual of the first adsorbent adsorbed onto the biomedical sample can be determined by measuring the intensity of the fluorescence. 
         [0036]    The first reagent-desorption step S 13  and the first discrimination step S 14  may be repeated several times and compared to achieve an optimal condition for detection. As described above, the amount of residual of the first adsorbent on the biomedical sample correlates to the length of time that a biomedical sample is immersed into the first desorbing reagent. The immersion time may be adjusted by repeating the first desorption step S 13  and the first discrimination step S 14  several times to determine cancer stage at different regions of biomedical sample. 
         [0037]    In one embodiment, a first pre-cleaning step may be performed prior to the first adsorption step S 12 , during which the biomedical sample is immersed into the first desorption reagent for a longer period of time than the predetermined period of time for removing impurities on the biomedical sample. 
         [0038]    Referring to  FIG. 2 , a method for detecting cancer according to another preferred embodiment of the present invention is provided. The method, in addition to the steps illustrated in  FIG. 1 , further comprises the following steps. A second reagent-adsorption step S 21  is performed during which the biomedical sample is immersed into a second detection reagent comprising a second adsorbent that would be physically adsorb onto the biomedical sample. Next, a second desorption step S 22  is performed, which includes immersing the biomedical sample into a second desorption reagent for a second period of time. The second desorption reagent comprises an organic solvent for desorbing the second adsorbent adsorbed onto the biomedical sample. Next, a second discrimination step S 23  is performed, which includes measuring the amount of residual of the second adsorbent adsorbed on the biomedical sample for determining the distribution of the cancer cells within the biomedical sample. The second reagent-desorbing step S 22  and the second discrimination step S 23  may be repeated several times, and the results are compared with each other to optimize the detection procedures. 
         [0039]    As described above, the physisorption capability of the first adsorbent may be greater or less for cancer cells than that of normal cells in the biomedical sample. Therefore, in one preferred embodiment, two kinds of adsorbents with greater and less physisorption capability for the cancer cells than that the normal cells in the biomedical sample, respectively, are compared to determine the cancer cells distribution within the biomedical sample. 
         [0040]    Further to the above description, the physisorption capability of the first adsorbent and the second adsorbent for the cancer cells may be greater and less than that of the normal cells in the biomedical sample, respectively. For example, the first adsorbent comprises a long-chain ester containing 16-46 carbon atoms; and the second adsorbent comprises a long-chain alkane containing 21-30 carbon atoms. 
         [0041]    Alternatively, the physisorption capability of the first adsorbent for the cancer cells may be less and the second adsorbent may be greater than that of the normal cells in the biomedical sample, respectively. For example, the first adsorbent comprises a long-chain alkane containing 21-30 carbon atoms, and the second adsorbent comprises a long-chain ester containing 16-46 carbon atoms. 
         [0042]    In one embodiment, the method for cancer detection comprises a second pre-cleaning step prior to the second adsorption step S 21 , which includes immersing the biomedical sample into the second desorption reagent for a longer time than the second predetermined time for removing impurities on the biomedical sample. 
         [0043]    The present invention may be described in detail by the following embodiments, which are presented for the purposes of illustrations and descriptions, and they are not intended to limit the scope of the present invention. 
       Embodiment 1: Colon Cancer Detection 
       [0044]    This embodiment describes a method for determining colon cancer. 
         [0045]    1. First, a sample of cell lines is fixed on a slide and then immersed into Dimethylbenzene (Xylene, C 8 H 10 ) for 20 minutes. Next, the infrared spectrum and spectral image of the sample is captured as the background as follows for determining cancer. An organic solvent, for example but not limited to, xylene and a detection reagent at 25° C. are employed, and the sample is dried at room temperature. 
         [0046]    2. Next, the sample is immersed for 2 minutes into a xylene solution, C 25 H 52  containing 5 wt % paraffin which serves as the first adsorbent, and then the sample dried at room temperature. 
         [0047]    3. Next, the waxed sample of cell lines is immersed in xylene for 5 seconds to partially desorb the paraffin and then dried at room temperature. Next, the absorption spectra and spectral images of the waxed sample of cell lines fixed on a slide are taken. The infrared images of sample of cell lines adsorbing wax are obtained by measuring the absorbance of wax in the range of 3000-2800 cm −1 . 
         [0048]    4. The above step 3 is repeated thrice, by immersing the sample for 5, 10 and 15 seconds in xylene for completely desorbing paraffin. 
         [0049]      FIG. 3  illustrates the infrared spectral images of samples of human cell lines illustrating the results based on the physisorption capability of paraffin.  FIG. 3(   a ),  FIG. 3(   f ) and  FIG. 3(   k ) are visible light images of a sample of normal cell lines CCD- 18 Co, and two samples of cancer cell lines, SW- 480  and SW- 403 , respectively.  FIG. 3(   b ),  FIG. 3(   g ) and  FIG. 3(   l ) illustrate infrared spectral images of sample cell lines, CCD- 18 Co, SW- 480 , and SW- 403 , respectively, following treatment with xylene for 20 minutes and drying in air. These images serve as reference background images for samples of cell lines after paraffin adsorption and paraffin desorption.  FIG. 3(   c - e ),  FIG. 3(   h - j ) and  FIG. 3(   m - o ) are infrared spectral images of sample of cell lines, CCD- 18 Co, SW- 480  and SW- 403 , after paraffin desorption for 5 seconds, 10 seconds and 15 seconds, respectively. Comparing reference background images (i.e. the infrared spectral images of cell samples after cleaning with xylene for 20 minutes) with infrared spectral images of cell samples for different paraffin desorption time intervals, it is observed that the normal cells exhibit more absorbance in the range of 3000-2800 cm −1  because of paraffin residue. On the contrary, the paraffin adsorbed on cancer cell lines does not exhibit more absorbance in the range of 3000-2800 cm −1 . 
         [0050]    5. A second adsorption step using a second adsorbent, beeswax (C 46 H 92 O 2 ), is performed. The infrared absorption spectra of the sample cell lines is taken after cleaning with xylene for 20 minutes to remove adsorbed paraffin and then dry in air. An infrared spectral image of the sample of cell lines is thus obtained based on the spectra of sample of cell lines after cleaning with xylene for 20 minutes. 
         [0051]    6. A sample of cell lines on a slide is immersed for 2 minutes in a solution of xylene containing 5 wt % beeswax and then taken out of the solution and dried at room temperature. 
         [0052]    7. The infrared absorption spectrum and the infrared spectral image of the sample of cell lines are taken after cleaning with xylene for 5 seconds and then air-dried at room temperature, during which the sample of cell lines is beeswaxed and fixed on a slide and desorbed with xylene. 
         [0053]    8. The above step 7 is repeated thrice by immersing the sample for 5, 10 and 15 seconds in xylene to desorb completely beeswax. 
         [0054]      FIG. 4  illustrates the result of infrared spectral images illustrating the results of desorption of beeswax from the sample of cell lines.  FIG. 4(   a ),  FIG. 4(   f ) and  FIG. 4(   k ) are white light images of a sample of normal cell line, CCD- 18 Co, and two samples of cancer cell lines, SW- 480  and SW- 403 , respectively.  FIG. 4(   b ),  FIG. 4(   g ) and  FIG. 4(   l ) are infrared spectral images of sample of cell lines CCD- 18 Co, SW- 480 , and SW- 403 , respectively, after being cleaned with xylene for 20 minutes and air-dried, which is constructed based on the absorbance of lipid in the range of 3000-2800 cm −1 . These spectral images serve as reference background images for beeswaxing and debeeswaxing sample of cell lines.  FIG. 4(   c - e ),  FIG. 4(   h - j ) and  FIG. 4(   m - o ) illustrate the infrared spectral images of sample cell lines, CCD- 18 Co, SW- 480  and SW- 403  after debeeswaxing by immersing the sample for 5, 10 and 15 seconds. Comparing reference background images (i.e. the infrared spectral images of sample of cell lines after being cleaned for 20 minutes with xylene) with infrared spectral images of sample of cell lines after being cleaned for various time periods with xylene, it is observed that the infrared spectral images of cancer cell lines exhibit substantially higher in absorbance within the range of 3000-2800 cm −1  due to the beeswax adsorbed onto the membrane of cancer cell lines. However, the amount of residual of beeswax on the membrane of normal cell lines is rare. Therefore, the aforementioned method can be rapid and effective in differentiating normal cell lines from cancer cell lines, and can be applied in colorectal cancer detection. 
       Embodiment 2: Oral Cancer Detection (I) 
       [0055]    The aforementioned method may also be adopted in determining oral cavity cancer, wherein there are 5 wt % paraffin and 5 wt % beeswax in xylene using as the first adsorbent and the second adsorbent, respectively. 
         [0056]      FIG. 5  illustrates the result of infrared spectral images illustrating infrared adsorption of paraffined sample cell lines. Both  FIG. 5(   a ) and  FIG. 5(   f ) are white light images of samples of normal oral cavity cell lines, normal human keratinocytes (NHOK), and a sample of human oral cavity cancer cell lines, oral epidermoid carcinoma (OECM-1), respectively.  FIG. 5(   b ) and  FIG. 5(   g ) are infrared spectral images of sample cell lines NHOK and OECM-1 after being cleaned for 20 minutes with xylene and dried at room temperature, which are constructed based on the absorbance of lipid in the range of 3000-2800 cm −1 . These images serve as reference background images for paraffin adsorption and desorption capabilities.  FIG. 5(   c - e ) and  FIG. 5(   h - j ) are infrared spectral images of cell samples NHOK and OECM-1, respectively, after desorption of paraffin for various period of times. Obvious amount of residual of adsorbed paraffin on NHOK compared to that on OECM-1 is illustrated in  FIG. 5 . 
         [0057]    Additionally,  FIG. 6  illustrates the result of infrared spectral images illustrating beeswax adsorption of cell sample. Both  FIG. 6(   a ) and  FIG. 6(   f ) are white light images of samples of normal oral cavity cell lines NHOK and of oral cavity cell lines OECM-1, respectively.  FIG. 6(   b ) and  FIG. 6(   g ) are infrared spectral images of cell lines NHOK and OECM-1 after being cleaned for 20 minutes with xylene and then dried, and the infrared spectral images are established based on the absorbance in the range of 3000-2800 cm −1 . These images serve as reference background images for beeswax adsorption and beeswax desorption;  FIG. 6(   c - e ),  FIG. 6(   h - j ) are infrared spectral images of sample cell lines NHOK and OECM-1, respectively, after desorption of beeswax for various period of times. As can be seen in  FIG. 6 , the amount of residual of adsorbed beeswax on OECM-1 is substantially more than that on NHOK. 
       Embodiment 3: Oral Cancer Detection (II) 
       [0058]    The procedures for determining the oral cavity cancer according to this method is similar to the process steps 1-8 of embodiment 1 described above, except for the first adsorbent comprises 5 wt % Triacontane (C 30 H 62 ); the second adsorbent comprises 7.5 wt % ethyl myristate (C 16 H 32 O 2 ). 
         [0059]      FIG. 7  illustrates the result of infrared spectral images illustrating triacontane (C 30 H 62 ) adsorption onto sample cell lines. Oral squamous cancer cell (OECM-1) lines after adsorption and desorption of triacontane in xylene for 5 seconds also exhibit a characteristic absorption of adsorbed C 30 H 62  in the spectral range of 3000-2800 cm −1 . 
         [0060]      FIG. 8  illustrates the result of infrared spectral images illustrating ethyl myristate (C 16 H 32 O 2 ) adsorption onto sample cell lines. Normal human oral keratinocyte (NHOK) cell lines after adsorption and desorption of ethyl myristate in xylene for 5 seconds exhibit a stronger absorption of adsorbed C 16 H 32 O 2  in the spectral range 3000-2800 cm −1 . 
       Embodiment 4: Detection of Oral Cavity Cancer (III) 
       [0061]    The procedure for determining the oral cavity cancer (III) is similar to the process steps 1-4 of embodiment 1 described above, except for the first adsorbent comprises a xylene solution containing 10 wt % 3,5,5-Trimethylhexyl 3,5,5-trimethylcaproat (C 18 H 36 O 2 ). 
         [0062]      FIG. 9  illustrates the result of infrared spectral images illustrating 3,5,5-Trimethylhexyl 3,5,5-trimethylcaproat (C 18 H 36 O 2 ) adsorption onto sample cell lines. Normal human oral keratinocyte (NHOK) cell lines after adsorption and desorption of 3,5,5-Trimethylhexyl 3,5,5-trimethylcaproat in xylene for 5 seconds exhibits a stronger absorption of adsorbed C 20 H 40 O 2  is in the spectral range 3000-2800 cm −1 . 
       Embodiment 5: Detection of Oral Cavity Cancer Tissue 
       [0063]    A method for determining the oral cavity cancer tissue may be described as follows. 
         [0064]    1. A paraffin-embedded or other material-embedded tissue section sample is fixed on a slide and a suitable solvent is employed to remove embedded material from tissue section sample. After removing the embedded material, and the tissue section sample is cleaned for 60 minutes with xylene to ensure that no residual of embedded material within tissue section before detecting cancer. Finally, infrared spectra of the tissue section sample and the infrared spectral image are obtained to serve as reference background for the following procedures. 
         [0065]    2. A tissue section sample is immersed for 10 minutes in a xylene solution containing 5 wt % paraffin as a first adsorbent, and then the tissue section sample is taken out of xylene solution and dried at room temperature. 
         [0066]    3. Next, the sample is washed for 30 seconds with xylene and then dried at room temperature, and spectra and spectral images of the tissue section sample are obtained. 
         [0067]    4. The above step 3 is repeated twice with 30 and 60 seconds paraffin desorption time periods using xylene solution containing 5 wt % paraffin. 
         [0068]      FIG. 10  illustrates the result of spectral images illustrating lipid adsorption onto a paraffined tissue section sample.  FIG. 10(   a ) and  FIG. 10(   e ) are white light images of a normal oral cavity tissue section and an oral cavity cancer tissue (T2N1G1) section sample of human, respectively.  FIG. 10(   b ) and  FIG. 10(   f ) are infrared spectral images of a normal oral cavity tissue section and the oral cavity tissue (T2N1G1) section sample after being cleaned for 60 minutes with xylene and dried at room temperature, and serve as reference background images for adsorption and desorption of paraffin, which are based on the absorbance within the spectral range of 3000-2800 cm −1 .  FIG. 10(   c ) and  FIG. 10(   g ) are infrared spectral images of a normal oral cavity tissue section and a oral cavity cancer tissue section sample, respectively, after being cleaned for 30 seconds with xylene to desorb paraffin  FIG. 10(   d ) and  FIG. 10(   h ) are infrared spectral images of a normal oral cavity tissue section and the oral cavity cancer tissue section, respectively, after being treated for 60 seconds with xylene to desorb paraffin. 
         [0069]    Comparing the reference background images with the infrared spectral images of oral cavity tissue section samples after desorbing paraffin for variant period of time, the normal tissue section sample illustrates a higher absorbance in the range of 3000-2800 cm −1  due to adsorbed paraffin, on the contrary, the adsorbance due to adsorbed paraffin on a given oral cavity cancer tissue section sample in the range of 3000-2800 cm −1  is not significant. 
         [0070]    5. A spectrum of a given paraffined oral cavity tissue section sample after being cleaned for 60 minutes with xylene and dried at room temperature is obtained. 
         [0071]    A spectrum of a given sample of oral cavity tissue section is obtained, which is waxed with second adsorbent, beeswax, after being cleaned for 60 minutes with xylene to remove all of the beeswax and dried in air at room temperature. 
         [0072]    6. A second adsorbent, 5 wt % beeswax in xylene solution, is dripped on a tissue section sample and then dried the tissue section sample in air for 60 minutes at room temperature. 
         [0073]    7. An infrared spectrum of a beeswaxed tissue section sample is obtained after being cleaned for 30 seconds with xylene and dried in air at room temperature. 
         [0074]    8. The above step 7 is repeated twice with 30 and 60 seconds debeeswaxing process using a xylene solvent. 
         [0075]      FIG. 11  illustrates the result of spectral images illustrating beeswax adsorption onto a tissue section sample. Both  FIG. 11(   a ) and  FIG. 11(   e ) are white light images of a normal oral cavity tissue section and an oral cavity tissue section T2N1G1 of human, respectively.  FIG. 11(   b ) and  FIG. 11(   f ) illustrate the result of infrared spectral images of a normal oral cavity tissue section and an oral cavity cancer tissue (T2N1G1) section in the spectral range of 3000-2800 cm −1  after being cleaned for 20 minutes with xylene and dried in air at room temperature. These images serve as reference background images for beeswaxing and debeeswaxing of tissue section sample.  FIG. 11(   c ) and  FIG. 11(   g ) are infrared spectral images of a normal oral cavity tissue and an oral cavity cancer tissue section, respectively, after being cleaned for 30 seconds with xylene to desorb beeswax.  FIG. 10(   d ) and  FIG. 10(   h ) are infrared spectral images of a normal oral cavity tissue section and an oral cavity cancer tissue section after being treated for 60 seconds with xylene, respectively. 
         [0076]    Comparing the reference background images with the infrared spectral images of oral cavity tissue sections after debeeswaxing for various period of time intervals, it is observed that the oral cavity cancer tissue section exhibits a higher absorbance in the spectral range 3000-2800 cm −1  due to the residual of beeswax. On the contrary, the absorbance of beeswax adsorbed onto the normal oral cavity tissue section is low. Therefore, the said method can be applied to rapidly and effectively differentiate malignant part from a biomedical tissue, and can be applied to detect oral cavity cancer and colon cancer. 
       Embodiment 6: Cancer Diagnosis utilizing Florescence Microscopy 
       [0077]    1. First, a sample of cell lines is fixed on a slide and then cleaned for 20 minutes with xylene, and measured by an optical microscope. 
         [0078]    2. A reagent of dye-beeswax-xylene solution containing 0.01 wt % dye and 5 wt % is used for differentiating cancer cell lines from normal cell lines. The sample is immersed in dye-beeswax-xylene solution for 1 minute and then dried in air at room temperature. 
         [0079]    3. Fluorescence images of a sample of cancer cell lines are obtained after being cleaned for 5 seconds in pure xylene and dried in air. 
         [0080]    4. The above step 3 is repeated twice with 5 and 10 seconds of debeeswaxing process with xylene solvent. 
         [0081]      FIG. 12  illustrates the results of fluorescence images illustrating beeswax adsorption onto the sample of cell lines.  FIG. 12(   a ) is a white light image of cancer cell lines of SW- 403 .  FIG. 12(   b ) illustrates a control image without debeeswaxing and those images of debeeswaxed cancer cell lines of SW- 403  after being debeeswaxed for a duration of 5, 10, 15 and 60 seconds, respectively.  FIG. 12(   c ) illustrates an image without being deparaffined and those images of cancer cell lines of SW- 403  after being deparaffined for a duration of 5, 10, 15 and 60 seconds, respectively. The result showed that stronger intensity of fluorescence is exhibited by the beeswax adsorbed onto the sample of cancer cell lines 
         [0082]    In summary, a method for cancer detection of the present invention is based on the physisorption capability of detection reagent onto samples of cancer cell lines and normal cell lines. The method employing the said principle can be a rapid and non-invasive method, and sample can also be re-examined for further medical inspection. 
         [0083]    While the invention is susceptible to various modifications and alternative forms, a specific example thereof has been shown in the drawings and is herein described in detail. It should be understood, however, that the invention is not to be limited to the particular form disclosed, on the contrary, the invention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the appended claims.