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Methods for Determining Cancer Incidence or Risk of Cancer Incidence

A cancer, risk technology, used in disease diagnosis, material excitation analysis, nanotechnology for sensing, etc., can solve problems such as low detection sensitivity

Inactive Publication Date: 2016-02-03
TOHOKU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the detection sensitivity is low, and there are also quantitative problems

Method used

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  • Methods for Determining Cancer Incidence or Risk of Cancer Incidence
  • Methods for Determining Cancer Incidence or Risk of Cancer Incidence
  • Methods for Determining Cancer Incidence or Risk of Cancer Incidence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Preparation method of immunostained tissue sample

[0088] In order to distinguish the difference from HercepTest (breast cancer tissue detected by HER2 antibody), all HercepTest-negative tissue samples were used as human breast cancer pathological tissue. On the other hand, there are cancer cases unrelated to the three main factors (ER, PgR, HER2) related to the proliferation of breast cancer. It is generally believed that the "triple negative" breast cancer has a poor prognosis and is difficult to treat. Here, "triple negative" breast cancer tissue samples are also discussed. Specifically, 4 test samples of breast cancer tissues from recurrent patients (relapse within about 1 year and death within 4 years) (one of the test samples was triple negative [because 20-25% of recurrent patients are "triple negative", so random Add the "triple negative" label to 1 of the 4 test samples]), "triple negative" breast cancer tissue from recurrent patients (recurrence within 4 yea...

Embodiment 2

[0090] Problems with fluorescent immunohistostaining

[0091] For the immunostained tissue sample prepared in Example 1 above, a combination of a confocal unit (manufactured by Yokogawa Electric Corporation), a fluorescence microscope (manufactured by Olympus Corporation), and an electron multiplier CCD (EM-CCD) imaging device (Andor (manufactured by the company) was irradiated with excitation light at 488 nm, and then a fluorescence image (fluorescence still image) of quantum dot fluorescent particles (detected by PAR1ab-QD705) was obtained using a bandpass filter of 695 to 740 nm. figure 1 (b) shows an example of a fluorescence still image of a tissue sample of a patient with recurrent breast cancer. Autofluorescence is so strong that quantum dot fluorescent particles (detected by PAR1ab-QD705) cannot be detected at first glance. Actually, when comparing the fluorescence brightness of autofluorescence and quantum dot fluorescent particles (detected by PAR1-antibody), only a...

Embodiment 3

[0094] Creation of corrected fluorescence images with autofluorescence removed

[0095] Fluorescence still images with zero (0) fluorescence intensity of autofluorescence as background are required. In this way, the total fluorescence in the fluorescence still image can be calculated as the fluorescence derived from the quantum dot fluorescent particles. The contrast adjustment method of the above-mentioned embodiment 2 is a division method of fluorescence intensity, and zero (0) cannot be obtained by division, so it must be an image processing method using a subtraction method that can obtain zero (0). Thus, as an image processing method for removing autofluorescence in a fluorescent still image, the method described below is considered. First, the breast cancer tissue sample immunostained with quantum dot fluorescent particles is irradiated with excitation light (laser) with an excitation wavelength of 488 nm, and a fluorescence image of quantum dot fluorescent particles is...

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Abstract

The present invention provides a high-accuracy and quantitative determination method of cancer onset or cancer risk by performing a quantitative tissue staining method using, for example, an inhibitor of cancer cells in living tissue. Motility and invasive PAR1 antibodies and other antibodies that recognize cancer growth control factors or metastasis control factors. The present invention uses the following tissue staining method to determine the incidence of cancer or the risk of cancer incidence. The tissue staining method includes the following steps: labeling an antibody that recognizes a cancer proliferation control factor or metastasis control factor with a fluorescent substance, making the fluorescently labeled antibody A step of contacting a tissue sample; a step of irradiating excitation light to the tissue site in contact with the antibody to obtain a fluorescent image; obtaining fluorescence emitted by the fluorescent substance in the same field of view and at the same focus as the fluorescent image A step of obtaining an autofluorescence image in a region near the short wavelength side or the long wavelength side compared to the wavelength region; performing image processing, removing the fluorescence brightness of the autofluorescence image from the fluorescence brightness of the fluorescence image, and obtaining a corrected fluorescence image The process; the process of counting the number of cells in the tissue site contacted by the antibody; the process of calculating and measuring the average fluorescence brightness of one fluorescent particle; the process of calculating the number of fluorescent particles per one cell.

Description

technical field [0001] The present invention relates to a method for determining the onset of cancer or the risk of developing cancer, using a tissue staining method that effectively eliminates the influence of autofluorescence and enables quantitative analysis with high precision. Background technique [0002] Cancer, together with vascular system diseases represented by myocardial infarction and cerebral infarction, are the two major causes of death in adults. For example, although the incidence rate of breast cancer in Japan is lower than that of European and American countries, it tends to increase in recent years. In 1998, it surpassed the incidence rate of gastric cancer and became the first female incidence rate. According to the statistics of the Ministry of Health, Labor and Welfare of Japan in 2005, which is the most recent report, the annual number of breast cancer patients exceeds 50,000. [0003] In the diagnosis of cancer, in addition to image diagnosis such a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N21/64G01N33/48G01N33/53G01N33/536
CPCG01N21/6428G01N33/57415G01N2800/50G01N21/6458G01N33/588B82Y15/00G01N21/6486
Inventor 权田幸祐宫下穰武田元博大内宪明
Owner TOHOKU UNIV
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