CN105463054A - Method for calculating inhibition ratio of tumor medicine sensitivity test - Google Patents
Method for calculating inhibition ratio of tumor medicine sensitivity test Download PDFInfo
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- CN105463054A CN105463054A CN201610063897.1A CN201610063897A CN105463054A CN 105463054 A CN105463054 A CN 105463054A CN 201610063897 A CN201610063897 A CN 201610063897A CN 105463054 A CN105463054 A CN 105463054A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5082—Supracellular entities, e.g. tissue, organisms
- G01N33/5088—Supracellular entities, e.g. tissue, organisms of vertebrates
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Abstract
The invention discloses a method for calculating the inhibition ratio of a tumor medicine sensitivity test. The method includes the following steps of taking a black-white image of a tumor tissue block through transmitted-light illumination before medicine is added for acting, and subtracting the values of pixel luminance of the tumor tissue block from the average luminance of background pixels for summation to obtain the index A for reflecting the volume of the tumor tissue block unprocessed by medicine, adding MTT for cultivation after adding medicine for acting, taking a black-white image of the tumor tissue block through diffused illumination, subtracting the values of pixel luminance of the tumor tissue block from the average luminance of the background pixels for summation to obtain the index B for reflecting the activity of the tumor tissue block processed by medicine, setting a blank contrast tissue block during the test, and calculating the inhibition ratio of medicine for tumors. By means of the medicine, when the inhibition ratio is calculated, the two factors, namely, the thickness and the blue color shade, related to the activity, of the tumor tissue block are considered, and the problem that the accuracy of inhibition ratio calculation is limited in the tumor medicine sensitivity test method is solved.
Description
Technical field
The invention belongs to biomedicine field, particularly a kind of tumour medicine sensitivity test inhibiting rate method of calculation.
Background technology
Medical research and Clinical Laboratory relate to mensuration tumour to the susceptibility of various antitumor drug, for selection of clinical antitumor drug provides foundation.The present inventor discloses a kind of tumour medicine sensitivity test image analyzer camera head and Tumor Drug Sensitivity Test Method at utility model patent " tumour medicine sensitivity test image analyzer camera head " (CN200520053379.9).This method tumor tissue is placed in by stainless steel stent rest and the filter paper being in nutrient solution and Air Interface is cultivated, transillumination and the image with the computerized image analysis sgstem picked-up tumor tissue with camera is adopted before adding drug effect, binary conversion treatment is done to image, calculate the area of tumor tissue, add MTT after adding drug effect again and do short period of time cultivation, yellow MTT is reduced to the part tissue indigo plant dye that blue product makes the cell containing survival by the reductase enzyme of survivaling cell, adopt diffuse illumination and the indigo plant dye image with image analyzer picked-up tumor tissue, the blued area that binary conversion treatment calculates tumor tissue is done to image, inhibiting rate is calculated with following formula (1), determine the restraining effect of drug on tumor cell.
formula (1)
This method binaryzation tumor tissue area and blued area calculate the inhibiting rate of tumor tissue, are ignoring tumor tissue's thickness factor relevant with activity with the blue depth, thus accuracy system.
Summary of the invention
In order to overcome the shortcoming and deficiency that calculate inhibiting rate accuracy in prior art in Tumor Drug Sensitivity Test Method, the object of the present invention is to provide a kind of tumour medicine sensitivity test inhibiting rate method of calculation.The method is that a kind of factor relevant with activity for tumor tissue's thickness and the blue depth these two being taken into account when calculating inhibiting rate is improved one's methods.
Object of the present invention is achieved through the following technical solutions:
A kind of tumour medicine sensitivity test inhibiting rate method of calculation, comprise the steps:
(1) black white image of transillumination picked-up tumor tissue is adopted before adding drug effect, background pixel mean flow rate is deducted respectively to the value summation of each pixel intensity of tumor tissue, obtain the Index A of a reflection drug treating pre-neoplastic tissue block volume;
(2) after adding drug effect, add MTT again and do short period of time cultivation, adopt the black white image of diffuse illumination picked-up tumor tissue, background pixel mean flow rate is deducted respectively to the value summation of each pixel intensity of tumor tissue, obtain the index B of activity of tumor tissue after a reflection drug treating;
(3) blank tissue block is set up in test simultaneously, and is calculated the inhibiting rate of drug on tumor by formula 2 with the Index A of step (1) and the index B of step (2);
formula 2.
The calculation formula of Index A or B can be but is not limited to formula 3:
formula 3;
Wherein, L in formula 3
mfor background pixel average brightness, L
(x, y)for the brightness of each pixel of image, N and M is respectively the horizontal and vertical pixel count of image.
Tumor tissue is placed in by stainless steel stent rest and the filter paper being in nutrient solution and Air Interface is cultivated by the present invention, under transillumination condition, absorb the black white image of tumor tissue with tumour medicine sensitivity test image analyzer camera head before adding drug effect, by video signal cable, picture signal is sent to the computer that is equipped with image card and image analysis software, 3 pairs of images carry out computational analysis with the formula, background pixel mean flow rate is deducted respectively to the value summation of each pixel intensity of tumor tissue, obtain the Index A of a reflection drug treating pre-neoplastic tissue block volume, L in formula
mfor background pixel average brightness, L
(x, y)for the brightness of each pixel of image, the brightness of each pixel reflects the thickness of this tissue block, and the brightness of pixel and the thickness of tissue block are inversely proportional to, N and M is respectively the horizontal and vertical pixel count of image.After adding drug effect, then add MTT and do the short period of time and cultivate, yellow MTT is reduced to the part tissue indigo plant dye that blue product makes the cell containing survival by the reductase enzyme of the cell of survival, the blue degree that contaminates and area and viable cell number relevant.Under diffuse illumination condition, the image of tumor tissue's indigo plant dye is absorbed with tumour medicine sensitivity test image analyzer camera head, the brightness of each pixel reflects the activity of this tissue block, the brightness of pixel and the activity of tissue block are inversely proportional to, 3 pairs of images carry out computational analysis with the formula, background pixel mean flow rate is deducted respectively to the value summation of each pixel intensity of tumor tissue, obtain the index B of tumor tissues activity after a reflection drug treating.Blank tissue block is set up in test simultaneously, adopts formula 2 to calculate the inhibiting rate of drug on tumor.
The present invention, relative to prior art, has following advantage and effect:
Factor relevant with activity for tumor tissue's thickness and the blue depth these two is taken into account when calculating inhibiting rate by method of the present invention, overcomes in Tumor Drug Sensitivity Test Method the problem calculating inhibiting rate accuracy.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Embodiment 1
It is below the example of a liver-cancer medicine sensitivity test.The 24 every holes of well culture plate add the nutrient solution (DMEM substratum) of 1mL.The tissue block that liver cancer sample is cut into diameter 0.5 ~ 1mm is positioned on the filter paper of culture hole, 4 ~ 5 pieces, every hole, individual layer discharge is concentrated to form the tissue block of the thick 0.5 ~ 1mm of a slice diameter 1.5 ~ 2mm, cultivate after 1 day, porous culture plate is put into the image that Mei Kong tumor tissue taken by tumour medicine sensitivity test image analyzer camera head under transmitted light irradiates, calculate the Index A of tumor tissue with formula 3.Then every hole adds medicine 10 μ L, and often kind of medicine 4 repeating holes, medicine final concentration is cis-platinum (DDP) 12.5mgmL
-1, pidorubicin (EADM) 11.5mgmL
-1, 5 FU 5 fluorouracil (5-FU) 50.0mgmL
-1, mitomycin (MMC) 3.75mgmL
-1with bleomycin (BLM) 15mgmL
-1, 4 every holes of blank culture hole are separately set and add physiological saline 10 μ L.Cultivate after 4 days, it is 10mgmL that every hole adds concentration
-1mTT25 μ L cultivate 4 hours, porous culture plate is put into the image that every hole tissue block indigo plant dye taken by tumour medicine sensitivity test image analyzer camera head under diffuse illumination, calculates the index B of the blue dye part of tumor tissue with formula 3.Inhibiting rate is calculated with formula 2.The inhibiting rate of each medicine of result is respectively DDP93.4%, EADM48.9%, 5-FU18.2%, MMC91.0% and BLM39.5%.
formula 3;
Wherein, L in formula 3
mfor background pixel average brightness, L
(x, y)for the brightness of each pixel of image, N and M is respectively the horizontal and vertical pixel count of image.
formula 2.
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from spirit of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (2)
1. tumour medicine sensitivity test inhibiting rate method of calculation, is characterized in that comprising the steps:
(1) black white image of transillumination picked-up tumor tissue is adopted before adding drug effect, background pixel mean flow rate is deducted respectively to the value summation of each pixel intensity of tumor tissue, obtain the Index A of a reflection drug treating pre-neoplastic tissue block volume;
(2) after adding drug effect, add MTT again to cultivate, adopt the black white image of diffuse illumination picked-up tumor tissue, background pixel mean flow rate is deducted respectively to the value summation of each pixel intensity of tumor tissue, obtain the index B of activity of tumor tissue after a reflection drug treating;
(3) blank tissue block is set up in test simultaneously, and is calculated the inhibiting rate of drug on tumor by formula 2 with the Index A of step (1) and the index B of step (2);
formula 2.
2. tumour medicine sensitivity test inhibiting rate method of calculation according to claim 1, is characterized in that: described Index A or the calculation formula of B are formula 3:
formula 3;
Wherein, L in formula 3
mfor background pixel average brightness, L
(x, y)for the brightness of each pixel of image, N and M is respectively the horizontal and vertical pixel count of image.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002312761A (en) * | 2001-04-12 | 2002-10-25 | Matsushita Electric Ind Co Ltd | Image processing method for cell image |
| CN2780773Y (en) * | 2005-01-05 | 2006-05-17 | 中山大学 | Camera device for tumour medicine sensitivity testing image analyzer |
| CN103088102A (en) * | 2013-01-09 | 2013-05-08 | 苏州麦克威尔生物医药科技有限公司 | Sterile instant type medicine film and tumor drug sensitivity testing application thereof |
| CN103966168A (en) * | 2014-05-19 | 2014-08-06 | 广州恒迪生物科技有限公司 | Tumor living tissue in-vitro culture system and culture method |
-
2016
- 2016-01-29 CN CN201610063897.1A patent/CN105463054A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002312761A (en) * | 2001-04-12 | 2002-10-25 | Matsushita Electric Ind Co Ltd | Image processing method for cell image |
| CN2780773Y (en) * | 2005-01-05 | 2006-05-17 | 中山大学 | Camera device for tumour medicine sensitivity testing image analyzer |
| CN103088102A (en) * | 2013-01-09 | 2013-05-08 | 苏州麦克威尔生物医药科技有限公司 | Sterile instant type medicine film and tumor drug sensitivity testing application thereof |
| CN103966168A (en) * | 2014-05-19 | 2014-08-06 | 广州恒迪生物科技有限公司 | Tumor living tissue in-vitro culture system and culture method |
Non-Patent Citations (2)
| Title |
|---|
| 实现肿瘤化疗药物个体化的研究进展: "实现肿瘤化疗药物个体化的研究进展", 《2012医学科学前沿暨第二届个体化治疗与抗肿瘤药物研究新趋向研讨会论文集》 * |
| 梁永钜等: "一种新的组织块培养肿瘤药敏试验法", 《中国新药杂志》 * |
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Application publication date: 20160406 |