WO2006079143A1 - Detection du cancer - Google Patents

Detection du cancer Download PDF

Info

Publication number
WO2006079143A1
WO2006079143A1 PCT/AU2005/001615 AU2005001615W WO2006079143A1 WO 2006079143 A1 WO2006079143 A1 WO 2006079143A1 AU 2005001615 W AU2005001615 W AU 2005001615W WO 2006079143 A1 WO2006079143 A1 WO 2006079143A1
Authority
WO
WIPO (PCT)
Prior art keywords
tissue
collagen
image
tumour
matrix
Prior art date
Application number
PCT/AU2005/001615
Other languages
English (en)
Inventor
Sarah Pearson
Original Assignee
The University Of New England
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from AU2004906531A external-priority patent/AU2004906531A0/en
Application filed by The University Of New England filed Critical The University Of New England
Publication of WO2006079143A1 publication Critical patent/WO2006079143A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence

Definitions

  • the invention relates to using second harmonic generation signals for detecting cancer, especially breast cancer.
  • Optical second-order harmonic generation (herein “SHG”) is a second-order nonlinear optical process, the signal being produced by constructive interference in non- symmetric environments. Because of this constraint it is ideally suited to imaging surfaces, interfaces and other mediums without inversion symmetry, such as chiral molecules.
  • US patent no. 6,208,886 relates to forming signal intensity maps from the application of SHG signals to tissue to resolve symmetry and content properties of layers in biological tissues.
  • a collagen matrix that is characteristic of a tumour tends to be associated with collagen that has a straight conformation.
  • the median curvature of this straight collagen is 0, with a range of 0 to 0.1, as opposed to that of the typically curly collagen in normal tissue which has a median value of 0.32, range of 0.2 to 0.56.
  • the "curvature" of a collagen fibre is expressed as the height of a curve of a collagen fibre divided by width of the curve.
  • the method includes a step of assessing from the image whether the matrix at the tissue region is composed predominantly of collagen that has a straight conformation, with curvature generally less than 0.2.
  • the collagen has a curvature of about 0.1, although it may be less than 0.1, for example, between about.0 and 0.1.
  • the collagen characteristic of a tumour has also been found to be typically organised into thinner fibres than those in normal tissue.
  • Fibre diameters in malignant tissue range from 0.18 ⁇ m to 0.55 ⁇ m, with a median of 0.320 ⁇ m, whereas fibres from normal tissue have diameters ranging from 0.24 ⁇ m to 1.03 ⁇ m, with a median value of 0.445 ⁇ m.
  • the larger diameter fibres in normal tissue suggests that these fibres constitute a greater range in the number of fibrils in a fibre (up to 11) than for those studied in malignant tissue (up to 8).
  • the method includes a step of assessing from the image whether the matrix at the tissue region is composed predominantly of collagen that is organised into thin fibres. These fibres typically have a diameter of at least about 0.18 ⁇ m and generally less than 0.55 ⁇ m.
  • the diameter of collagen fibres may be up to 30% more than the values discussed above.
  • the method may include the step of assessing from the image whether the collagen fibres have a diameter of at least about 0.234 ⁇ m and generally less than 0.715 ⁇ m.
  • the inventor has also found that fibrils from malignant tissue have a smaller diameter than those present in normal tissue. Fibrils in normal tissue show a median of 0.05 ⁇ m, and range of 0.03 ⁇ m to 1.0 ⁇ m, whereas fibrils in malignant tissue have a median of 0.04 ⁇ m, range 0.04 ⁇ m to 0.05 ⁇ m.
  • the method includes the step of assessing from the image whether the matrix at the tissue region is composed predominantly of collagen that is organised into fibrils that have a diameter of at least about 0.04 ⁇ m and generally less than 0.05 ⁇ m.
  • the diameter of collagen fibrils may be up to 30% more than the values discussed above.
  • the method may include the step of assessing from the image whether the collagen fibrils have a diameter of at least about 0.052 ⁇ m and generally less than
  • the above described method is particularly useful for determining whether breast tissue, including inter- and intra-lobular connective tissue, glandular and ductal tissue, includes a tumour.
  • a tumour is a cancer, neoplasm or other form of tissue that is characterised by a loss of normal control of tissue proliferation and/ or differentiation.
  • a tumour may be a single cell or multi- cellular mass.
  • the method is particularly useful for- the detection of carcinoma, especially invasive and non invasive carcinoma.
  • non malignant carcinoma examples include intraductal carcinoma, lobular carcinoma in situ and intraductal papillary carcinoma, benign Phyllodes Tumour.
  • invasive carcinoma examples include invasive ductal carcinoma, Paget's disease, invasive lobular carcinoma, medullary carcinoma, mucinous carcinoma, tubular (well- differentiated) carcinoma, invasive papillary carcinoma, adenoid cystic carcinoma, secretory carcinoma, apocrine carcinoma, carcinoma with metaplasia, malignant Phyllodes carcinoma, and mixed type carcinoma.
  • breast disease examples include benign neoplasia of the breast, such as fibroadenoma, other neoplasms, including Phyllodes tumour, lipomas, hemangiomas, leiomyomas, fibromas and angiosarcomas, sclerosing adenosis, hyperplasia, and non neoplastic diseases including fibrocystic disease.
  • benign neoplasia of the breast such as fibroadenoma, other neoplasms, including Phyllodes tumour, lipomas, hemangiomas, leiomyomas, fibromas and angiosarcomas, sclerosing adenosis, hyperplasia, and non neoplastic diseases including fibrocystic disease.
  • tissue examples include those having a connective tissue component that includes collagen, such as bowel, prostate, skin, cervical, ovarian tissue and the like.
  • tissue is selected from the group consisting of bowel, prostate, skin, cervix, ovarian tissue and the like. These and other examples of tissue may include epithelial cells. Other tissue such as cartilage and bone may also be determined according to the method.
  • the image may be assessed by assessing from the image whether the matrix at the tissue region is composed of collagen that predominantly has a straight conformation, as discussed above.
  • the image may be assessed by assessing from the image whether the matrix at the tissue region is composed of collagen that is organised into thin fibres, as discussed above. Also, the image may be assessed by assessing whether the matrix at the tissue region is composed of collagen that is organised into fibrils, the fibrils having a particular diameter, as discussed above.
  • the optical wave is typically a laser light.
  • the laser light has a wavelength in the range of about 700 to 900nm.
  • the second harmonic is typically in the range of about 350 to 450nm.
  • the laser light has a wavelength of about 830 nm and the second harmonic has a wavelength of about 410 to 415 nm. It will be understood that a laser light and second harmonic may have a wavelength that is more or less than those wavelengths described herein.
  • Any device for production of an optical wave having a wavelength as described herein may be used. Particularly preferred are those devices that are compatible with an apparatus for application of the optical wave to a tissue and/or for forming an image from the second harmonic (i.e the SHG signal) formed by application of the optical wave.
  • the tumour is detected in situ for example by using an optical fibre to apply an optical wave to a tissue.
  • an optical fibre to apply an optical wave to a tissue.
  • the method is performed in vitro.
  • a method for determining whether a tissue includes a tumour including:
  • the image is assessed by assessing from the image whether the matrix at the tissue region is composed of collagen that predominantly has a straight conformation; or collage that is organised into thin fibres; or collagen that is organised into fibrils having a particular diameter, as discussed above.
  • collagen is typically expressed in a tissue in the form of a matrix, or in other words, an association, collection or network of like fibres, and sometimes other fibres such as elastin and keratin.
  • images of normal tissue formed from SHG signals tend to represent a collagen matrix that has a particular degree of organisation.
  • a matrix tends to have a curly or crimped appearance, and it is distributed more or less evenly throughout the tissue. Further, acini and alveoli are clearly defined.
  • images of abnormal tissue formed from SHG signals tend to represent a collagen matrix in which organisation is relatively diminished as tissue tends toward neoplasia.
  • collagen tends to have a predominantly stringy or straight structure and it tends to be distributed unevenly through the tissue.
  • alveoli and acinus structures are poorly represented in these images.
  • an image of a collagen matrix, formed according to the method, that is characteristic of a tumour tends to be one in which a collagen matrix is represented as a stringy and uneven distribution of collagen. Further, where the tissue is breast tissue, the image may further have a poorly defined acini and alveoli morphology.
  • the inventor has further found that the collagen fibril diameters, measured using the SHG images, may be used to distinguish between normal and malignant tissue.
  • the inventor has found that the collagen fibrils in malignant tissue are smaller in diameter than those in normal tissue, and that these fibrils correspond to immature collagen fibrils, whereas those in normal tissue correspond to a mixture of immature and mature fibrils. Further, as discussed herein, the inventor has found that an image of a collagen matrix of a malignant tumour can be distinguished from the image of a collagen matrix of a benign tissue. Specifically, it has been found that collagen of a matrix associated with a malignant tumour tends to be composed predominantly of collagen having a straight conformation, as defined above. Collagen of a matrix associated with a benign tumour tends to be composed of collagen having both straight and curly or crimped conformation, as defined above.
  • the image may be assessed by assessing from the image whether the matrix at the tissue region is composed of collagen that predominantly has a straight conformation, as discussed above.
  • the image may be assessed by assessing from the image whether the matrix at the tissue region is composed of collagen that is organised into thin fibres, as discussed above.
  • the image may be assessed by assessing whether the matrix at the tissue region is composed of collagen that is organised into fibrils, the fibrils having a particular diameter, as discussed above.
  • the image of the matrix formed from the SHG signal can be assessed by any method or instrument in which the information in the image can be presented so as to be visible.
  • An example of a suitable method and instrument is described further herein.
  • Some components of this instrument are: a pulsed laser source linked to a microscope; a sample stage; appropriate filters to isolate the SHG signal; a detector of suitable resolution; imaging software to collect the signal and form an image; software for measuring fibre/fibril diameters, and morphological parameters.
  • one particular advantage of the method is that it has a much higher sensitivity for the detection of abnormal or neoplastic tissue than conventional techniques, such as histological techniques.
  • conventional techniques such as histological techniques.
  • the inventor by assessment of an image formed from SHG signals, the inventor has detected a tumour in the form of a stringy, unevenly dispersed collagen matrix in tissue that was otherwise considered by conventional histological technique to be normal. This is a particularly important advantage in the detection of breast cancer as these cancers tend to metastasise by forming infiltrating fenestrations that are often difficult to detect by conventional techniques.
  • the method comprises the steps of:
  • images of abnormal tissue formed from SHG signals tend to represent a collagen matrix in which the directional distribution of collagen fibres is anisotropic and highly directional.
  • images of normal tissue formed from SHG signals tend to represent a collagen matrix in which the directional distribution of collagen fibres is highly isotropic.
  • a method for determining whether a tissue includes a tumour including:
  • Figure 1 SHG image of tumour site breast tissue.
  • Figure 2 SHG image of breast tissue at 6 mm from the tumour site.
  • Figure 8 Short profile plot through a long (200 ⁇ m) straight section of fibres in a densely packed region of malignant tissue.
  • Figure 9 Short profile plot through a long (130 ⁇ m) straight section of sparsely distributed fibres in malignant tissue.
  • Figure 10 Profile plot through a short straight section of fibre groups in normal tissue.
  • Figure 11. 110 ⁇ m profile plot through an area where straight fibre groups are sparsely distributed in malignant tissue.
  • Figure 12. 110 ⁇ m profile plot through an area where straight fibre groups are densely distributed in malignant tissue.
  • Figure 13 300 ⁇ m profile plot through an SHG image from malignant tissue showing a sparse, low intensity distribution.
  • Figure 14 300 ⁇ m profile plot through an SHG image from normal tissue showing a more even, higher intensity distribution.
  • Figure 15 Surface plots of a) malignant tissue SHG signal, b) normal tissue SHG signal, demonstrating the sparse distribution of collagen in malignant tissue versus the more even distribution in normal tissue.
  • Figure 16 Histogram of fibril diameters in malignant tissue.
  • Figure 17 Histogram of fibril diameters in normal tissue.
  • FIG. 18 SHG images of a) normal tissue, b) malignant tissue at the same PMT voltage settings, c) malignant tissue at different PMT settings, showing the presence of straight, fine fibres.
  • Figure 20 SHG maximum surface plots of a) normal tissue, at 20 0 C, and b) normal tissue at 60 0 C at the same PMT settings.
  • Figure 21 SHG average surface plots of a) normal tissue at 20 0 C, and b) normal tissue at 60 0 C at the same PMT settings.
  • Figure 22 SHG minimum surface plots of a) normal tissue at 20 0 C, and b) normal tissue at 60 0 C at the same PMT settings.
  • Figure 23 SHG images of a) malignant tissue with the same PMT settings as for figure 19, b) the same sample, the image having been collected at different PMT voltage settings.
  • Figure 24 High resolution image of fine fibres at the end of a thick fibre group (image is 58 ⁇ m by 30 ⁇ m).
  • Figure 26 a) H&E stained histopathology slide image of tissue in a benign area of tissue showing fibroblasts in a highly activated state , b) the SHG ' image of the same sample in this region.
  • Figure 27 a) H&E stained histopathology slide image of tissue in a malignant area of the same tissue sample shown in figure 19 showing fibroblasts in a highly activated state and b) the SHG image of the same sample in this region.
  • Figure 28 H&E stained histopathology slide image showing tumour cells forming along collagen fibres in Indian file in malignant tissue.
  • SHG imaging was conducted as discussed below using malignant tissue from 12 patients, benign tissue from 6 patients and normal tissue from 10 patients. In the order of 800 SHG images have been produced from these tissue samples.
  • the lesion types include: invasive ductal, ductal hyperplasia with atypia, fibroadenoma, ductal hyperplasia without atypia, sclerosis adenosis, intraduct papilloma, fibrofatty, and malignant tissue of grades 1 , 2 and 3. Classification rates of 100% were achieved when classifying the tissue as malignant, benign or normal using collagen fibre morphology and macroscopic tissue morphology, as discussed below, as measures of disease. In one case a 20 ⁇ m piece of malignant tissue was observed in a benign sample.
  • Laser light from a Coherent Mira tuneable pulsed titanium sapphire laser of wavelength 830 nm and pulsewidth approximately 100 fs, was utilised in order to produce SHG signals in human breast tissue.
  • the wavelength was checked with a Rees spectrum analyser and pulsewidth with an APE autocorrelator.
  • tissue samples imaged comprised human breast tissue sourced from biopsy samples as follows;
  • the equipment used to produce the SHG signals included a Leica DMIRBE inverted stand equipped with a Leica TCS2MP confocal system.
  • the microscope is equipped with dual photomultiplier transmitted light detectors.
  • a 415/10-nm narrow bandpass filter (with the laser tuned to 830 nm) was used to exclude fluorescent signals in the transmission detector.
  • the main objective used was a 25x NA 0.75 oil-immersion planapochromat and an oil-immersion condenser was also used.
  • the second objective used was a 40. OX/1.25 oil-immersion planapochromat.
  • the laser light was passed through the tissue sample, then filtered using the 415/10 nm narrow bandpass filter to select the SHG signals which were registered using photomultiplier detectors.
  • SHG images were collected automatically as frame-by-frame sequential series, the average of four images at the same position being used in order to reduce the effects of background signals.
  • Leica software was then utilised to form images using the SHG signals. Formation of each image
  • tumour tissue can be distinguished from that of normal tissue, and hence that tumour tissue can be detected, according to one or more of an assessment of: (i) an image formed from SHG signals; (ii) SHG signal anisotropy.
  • an image formed from SHG signals is the structure of collagen fibres, tissue morphology highlighted by collagen distribution, fibre curvature, fibre distribution, fibre/fibril diameters, and presence of immature collagen fibres.
  • each of the above parameters are markers for disease, in particular breast cancer, and hence represent a technique for the detection and diagnosis of cancer.
  • Figure 1 demonstrates the unique nature of the collagen structure at the tumour site. Note the 'stringy' nature of the collagen fibres in some areas of the image. Also note the uneven distribution of the collagen matrix, some areas of the image indicating a very sparse collagen presence.
  • Figure 2 shows a representative image generated by collagen at 6 mm from the tumour site. This tissue had been diagnosed as 'normal' tissue by the pathologist.
  • the collagen structure here differs greatly from that observed at the tumour site; the collagen matrix has a 'curly' or 'crimped' arrangement and its distribution, in the areas where acinus or alveoli are not present, is more uniform.
  • Figure 3 gives another example of the 'stringy' nature of the collagen matrix at the tumour site. This effect was observed in all tumour site images, irrespective of tissue morphology.
  • Figure 4 also indicates the presence of traces of 'stringy' collagen, normally associated with tumour site tissue, close to relatively well-defined alveoli. This is a significant observation, indicating the possibility that the method can image the production of 'stringy' collagen, created by the tumour to enable metastasis. The remainder of the image shows evenly distributed 'curly' collagen, associated with normal tissue, and the presence of well-defined alveoli.
  • Figure 2 for the case where normal tissue was imaged, the alveoli are well defined and circular. Since they are acini they do not contain collagen and hence are observed as black saclike structures.
  • Figure 1 indicates that the alveoli at the tumour site are poorly defined, showing infiltration by 'stringy' collagen and not possessing clear edges.
  • the distribution of the collagen matrix also differs between tissue states; the 'normal' tissue indicates an even distribution of collagen matrix throughout the tissue, where alveoli are not present, whereas the images from tumour tissue indicate sparse representation of collagen in some areas.
  • tissue necrosis causing destruction of tissue morphology, reduction in the collagen matrix, and the formation of new 'stringy' collagen as a path for metastasis.
  • This change in tissue morphology was observed to change gradually from the tumour site to 6 mm where the tissue morphology was very well defined. .
  • tissue morphology recognised using this technique includes adipose (figure 5), ductal carcinoma in situ (figure 6), and cysts (figure 7).
  • adipose (figure 5)
  • ductal carcinoma in situ (figure 6)
  • cysts (figure 7).
  • the presence of a thick layer of collagen around the cyst should be noted, this differentiating it from ductal carcinoma in situ.
  • Collagen fibre distribution was studied using the SHG images, and compared in straight fibre sections within both malignant and normal tissue.
  • Figures 8 and 9 show intensity profiles over a short region (15 ⁇ m) of the image for malignant tissue, figure 8 being for fibres in a densely packed region, figure 9 for sparsely distributed fibres. These figures indicate that densely packed straight fibres in malignant tissue exist in slightly thicker groupings than those in sparsely packed regions. When both figures 8 and 9 are compared with that for normal tissue (figure 10) it can be seen that the straight fibres in normal tissue are more densely packed and exist in thicker fibre groupings. Gray value comparisons cannot be made between the plots for malignant and normal tissue in this case as they were obtained using different voltage settings on the PMT.
  • Plot profiles were produced for SHG images from both malignant and normal tissue.
  • the diameters of fibres and individual fibrils were measured and comparisons made. It was found that the fibres from normal tissue had diameters ranging from 0.24 ⁇ m to 1.03 ⁇ m, with a median value of 0.445 ⁇ m and variance 0.034 ⁇ m.
  • fibres from normal tissue are found to be larger than those observed in malignant tissue, this being due either to collagen degradation or fibrillogenesis in the malignant tissue. It is suggested that the cause might be fibrillogenesis, this being discussed in the section "fibrillogenesis and degradation".
  • the larger number of measured diameters (representing 11 fibre diameter groupings, 11 peaks in a histogram of fibre diameters) in normal tissue suggests that these fibres constituted a greater range in the number of fibrils in a fibre than for those studied in malignant tissue (representing 8 fibre diameter groupings, 8 peaks in a histogram of fibre diameters).
  • fibrils in malignant tissue do not have diameters greater than 0.05 ⁇ m, whereas fibrils in normal tissue can have diameters of up to 0.1 ⁇ m.
  • the distribution of fibril diameters was found to be unimodal for malignant tissue (median 0.04 ⁇ m with a standard deviation of 0.005 ⁇ m, indicating that a unimodal description is appropriate), and multimodal for normal tissue (see figures 16 and 17).
  • Fibrils in both malignant and normal tissue have diameters that are multiples of 8 - 10 nm. This agrees with research conducted using small-angle x-ray scattering (SAXS) and electron microscopy 1 ' 2 ' 3 . It should be noted that these values may be reduced with respect to those for live tissue since the tissue preparation (fixation and embedding) can cause shrinkage or expansion of the structures under investigation 2 .
  • SAXS small-angle x-ray scattering
  • Fibrillogenesis & Degradation of collagen is expected to be observed in malignant tissue as the tumour cells cause necrosis and remodelling of the extracellular matrix (ECM) in order to invade the surrounding tissue. This is observed in our images by a reduction in SHG signal intensity from malignant tissue versus normal tissue, and also by a reduction in the number and density of collagen fibres observed in the image (as previously mentioned). This is demonstrated in figures 18 a) and b), taken at the same voltage across the PMT and hence directly comparable in terms of intensity. In addition to this, when the SHG image from malignant tissue is collected at PMT settings such that the collagen fibres present are highlighted, straight, fine fibres are observed that are not present in the images of normal tissue. This is shown in figure 18 c).
  • Normal breast tissue was subjected to degradation by heating the tissue to 60 0 C, this degradation being similar to proteolysis caused by enzymatic catalysis present in malignancy.
  • Small-angle x-ray scattering results also indicate that the collagen fibres unravel in the presence of malignancy, producing a larger scattering surface area for amorphous scatter 4 .
  • the SHG images obtained from this degraded tissue were analysed, the results indicating that the straight collagen observed in malignant tissue is not a product of degradation.
  • Figures 19 a) and b) show representative SHG images obtained for normal tissue at 20 0 C and 60 0 C respectively, taken with the same voltage settings on the PMT. From this it can be seen that heating the tissue to 60 0 C causes a small but noticeable degradation of collagen, which is translated into a decrease in the SHG signal.
  • the images are from different slices of the tissue sample and hence morphology is not directly matched, however, the morphology of the tissue remains largely unaffected by heating to 60 0 C.
  • the outline structure of the ducts is only slightly blurred on heating. It should also be noted that there are no straight, fine collagen fibres present in either case.
  • the surface plots given in figure 20 a) and b) show the maximum intensity distributions observed in a) normal tissue at 20 0 C, b) normal tissue at 60 0 C at the same PMT voltage as used to produce the images in figure 19.
  • the surface plots given in figure 21 a) and b) show the average intensity distributions observed in a) normal tissue at 20 0 C, b) normal tissue at 60 0 C at the same PMT voltage as used to produce the images in figure 19.
  • the surface plots given in figure 22 a) and b) show the minimum intensity distributions observed in a) normal tissue at 20 0 C, b) normal tissue at 60 0 C at the same PMT voltage as used to produce the images in figure 19, In all these comparisons it can be seen that the intensity distribution is greater for the tissue kept at 20 0 C than for that heated to 60 0 C.
  • the mean SHG intensity is calculated for images from tissue at both temperatures and a comparison made, it is found that the median of the mean SHG signal intensities for the normal tissue at 20 0 C is 36.5, and 21.8 for the tissue heated to 60 0 C.
  • Figure 23 shows the SHG image from a malignant tissue sample, taken at a) the same PMT settings as those used to produce figure 19, and b) different settings used to highlight the straight fibres.
  • Figure 23 a) indicates that collagen degradation in malignant tissue is much greater than that produced when heating tissue to 60 0 C.
  • a comparison of figure 23 b) with figure 19 b) also indicates that the straight collagen observed in malignant tissue is not observed in normal tissue degraded by heating.
  • the morphology of the malignant tissue also shows significant distortion, to a much greater extent than for the heated normal tissue. This is as expected since the presence of tumour cells can lead to both necrosis, regions where tumour cells have infiltrated normal tissue and mimicked ductal tissue, all of which will yield unevenly outlined areas where no collagen is present.
  • Fibril diameters in malignant tissue are significantly smaller than those in normal tissue.
  • the distribution of fibril diameters is unimodal in malignant tissue and multimodal in normal tissue.
  • Figure 26 a shows fibroblasts in a benign area of tissue, these fibroblasts being in a normal active state.
  • Figure 26 b) shows the SHG image of the same sample in this region. The image contains no straight collagen associated with malignancy, and would be classed as benign when observing the state of the collagen and tissue morphology.
  • Figure 27 a shows fibroblasts in a malignant area of tissue, these fibroblasts posses a dispersed chromatin, being in a highly active state.
  • Figure 27 b) shows the SHG image of the same sample in this region. The image contains a large amount of straight collagen associated with malignancy, and would be classed as malignant when observing the state of the collagen and tissue morphology. This suggests that the collagen observed in malignant tissue using SHG is newly synthesised collagen.
  • the final parameter studied was the anisotropy of the SHG signal.
  • the SHG polarization anisotropy can be used to determine the absolute orientation and degree of organisation of the collagen in tissues.
  • the anisotropy parameter, ⁇ describing the
  • Polarisation of the laser light was achieved using a linear polariser, and a rotatable polariser placed between the condenser and the PMT detector allowed the SHG signal to be separated into parallel and perpendicular components.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Medical Informatics (AREA)
  • Biophysics (AREA)
  • Pathology (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Physics & Mathematics (AREA)
  • Molecular Biology (AREA)
  • Surgery (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention concerne l'utilisation de signaux de génération de seconde harmonique permettant de détecter le cancer, en particulier, le cancer du sein.
PCT/AU2005/001615 2004-11-15 2005-10-19 Detection du cancer WO2006079143A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AU2004906531A AU2004906531A0 (en) 2004-11-15 Detection of cancer
AU2004906531 2004-11-15

Publications (1)

Publication Number Publication Date
WO2006079143A1 true WO2006079143A1 (fr) 2006-08-03

Family

ID=36739948

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/AU2005/001615 WO2006079143A1 (fr) 2004-11-15 2005-10-19 Detection du cancer

Country Status (1)

Country Link
WO (1) WO2006079143A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107941765A (zh) * 2017-11-16 2018-04-20 南方医科大学南方医院 胃癌切除标本的胃浆膜表面胶原组织评价方法
US10229488B2 (en) 2010-03-31 2019-03-12 Agency For Science, Technology And Research Method and system for determining a stage of fibrosis in a liver

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6208886B1 (en) * 1997-04-04 2001-03-27 The Research Foundation Of City College Of New York Non-linear optical tomography of turbid media
WO2003055379A2 (fr) * 2001-12-24 2003-07-10 Salafsky Joshua S Procede utilisant une technique optique non lineaire a surface selective en vue d'imager des echantillons biologiques

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6208886B1 (en) * 1997-04-04 2001-03-27 The Research Foundation Of City College Of New York Non-linear optical tomography of turbid media
WO2003055379A2 (fr) * 2001-12-24 2003-07-10 Salafsky Joshua S Procede utilisant une technique optique non lineaire a surface selective en vue d'imager des echantillons biologiques

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
GUO Y. ET AL.: "Subsurface tumor progression investigated by noninvasive optical second harmonic tomography", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, vol. 96, no. 19, 14 September 1999 (1999-09-14), pages 10854 - 10856 *
YASUI T. ET AL.: "Characterization of collagen orientation in human dermis by two-dimensional second-harmonic-generation polarimetry", JOURNAL OF BIOMEDICAL OPTICS, vol. 9, no. 2, 2004, pages 259 - 264 *
ZIPPEL W. ET AL.: "Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation", PROCEEDINGS OF THE NATIONAL ACADEMY OD SCIENCES OF THE UNITED STATES OF AMERICA, vol. 100, no. 12, 10 June 2003 (2003-06-10), pages 7075 - 7080 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10229488B2 (en) 2010-03-31 2019-03-12 Agency For Science, Technology And Research Method and system for determining a stage of fibrosis in a liver
CN107941765A (zh) * 2017-11-16 2018-04-20 南方医科大学南方医院 胃癌切除标本的胃浆膜表面胶原组织评价方法

Similar Documents

Publication Publication Date Title
AU2014234022B2 (en) Measurement of tissue structures
Nguyen et al. Intraoperative evaluation of breast tumor margins with optical coherence tomography
US6965793B2 (en) Method for Raman chemical imaging of endogenous chemicals to reveal tissue lesion boundaries in tissue
Marghoob et al. Instruments and new technologies for the in vivo diagnosis of melanoma
US7697576B2 (en) Cytological analysis by raman spectroscopic imaging
Taroni et al. Diffuse optical spectroscopy of breast tissue extended to 1100 nm
US20050250091A1 (en) Raman molecular imaging for detection of bladder cancer
AU2010247132B2 (en) Tissue sample analysis
Gubarkova et al. Tissue optical properties estimation from cross-polarization OCT data for breast cancer margin assessment
Ooi et al. Fourier transform infrared imaging and small angle x‐ray scattering as a combined biomolecular approach to diagnosis of breast cancer
Wang et al. Recent advances in spontaneous Raman spectroscopic imaging: instrumentation and applications
Anuthama et al. Characterization of different tissue changes in normal, betel chewers, potentially malignant lesions, conditions and oral squamous cell carcinoma using reflectance confocal microscopy: Correlation with routine histopathology
Mahmoud et al. Delineation and detection of breast cancer using novel label-free fluorescence
Canpolat et al. Intra-operative brain tumor detection using elastic light single-scattering spectroscopy: a feasibility study
WO2006079143A1 (fr) Detection du cancer
Sunder et al. Laser Raman spectroscopy: A novel diagnostic tool for oral cancer
EP2887050A1 (fr) Procédé de démarcation sans marqueur de tissus
Nguyen et al. Portable real-time optical coherence tomography system for intraoperative imaging and staging of breast cancer
Lin et al. Diagnosing pituitary adenoma in unstained sections based on multiphoton microscopy
WO2017005838A1 (fr) Examen non invasif de tissu biologique sur la base d'imagerie par tomographie en cohérence optique haute définition en champ plein
Boppart et al. Coherent optical imaging and guided interventions in breast cancer: translating technology into clinical applications
Gubarkova et al. Contrast Enhancement of Cross-Polarization OCT Images of Breast Cancer by Optical Coefficient Calculation
Wu et al. Application of a Novel Miniaturized Histopathologic Microscope for Ex Vivo Identifying Cerebral Glioma Margins Rapidly During Surgery: A Parallel Control Study
Koul et al. Breast Tumor Margin Assessment Using Sub-Terahertz Wave
Pearson et al. Small-angle X-ray scattering and second-harmonic generation imaging studies of collagen in invasive carcinoma

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application
NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 05856170

Country of ref document: EP

Kind code of ref document: A1