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Method for detecting asbestos

一种检测方法、石棉的技术，应用在测量装置、颜色/光谱特性测量、仪器等方向，能够解决无法快速检测石棉、无法应对石棉风险、难多个处理等问题，达到简便且高精度检测的效果

Inactive Publication Date: 2014-04-16
HIROSHIMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, observation with a phase-contrast microscope requires skilled skills and considerable time, so it is difficult to perform multiple treatments at the same time
In addition, electron microscopes are very expensive equipment, so not everyone can easily implement
In addition, the determination by the electron microscope requires not only complicated pretreatment of the sample, but also the analysis of the fibers observed by the phase contrast microscope with an energy dispersive X-ray analyzer, which requires a lot of time and patience.
[0007] Therefore, it is impossible to quickly detect asbestos in the existing method using a phase-contrast microscope and an electron microscope (hereinafter referred to as "phase-contrast microscope-electron microscope method")
Therefore, the risk of asbestos at the demolition site, which requires rapid detection, cannot be addressed

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0184] [Example 1: Asbestos binding protein H-NS 60-90 Preparation and fluorescent labeling]

[0185] The biotin-streptavidin interaction is used to combine the biotin-modified asbestos-binding protein with fluorescently labeled streptavidin to produce fluorescently-labeled asbestos-binding protein.

[0186] First, make biotinylated asbestos binding protein. Using protein expression vector pET21-b (Novagen) as a template, oligonucleotide primer P1 (GCTCAGAAAATCGAATGGCACGAACACCACCACCACCACCACTGAACTA: SEQ ID NO:1) and oligonucleotide primer P2 (CTCGAAGATGTCGTTCAGACCGCCACCCTCGAGTGCGGCCGCAAGCTTGTC: SEQ ID NO: 2) were used for reverse PCR , Thus insert the biotinylated tail before the HisTag of pET21-b. The reverse PCR reaction was performed using KOD-plus-mutagenic kit (TOYOBO) according to the company's protocol. The biotinylated protein expression vector was named "pET21-AviTag-C".

[0187] Then, using the genomic DNA of Escherichia coli K-12 (Escherichia coli K12, ATCC700926) as a t...

Embodiment 2

[0193] [Example 2: Asbestos Microscopic Observation by Phase Contrast Fluorescence Method (1)]

[0194] Cut the membrane filter containing asbestos amosite (thea asbestos) (JAWE231) as the test substance into 1 / 8 pieces, with the collecting side facing up, add 20μl of buffer C [0.3M phosphate buffer three times Solution (pH8.0), 0.3M NaCl, 0.5% Tween80 (registered trademark)].

[0195] Then, add 5 times 20μl of H-NS containing fluorescent protein 60-90 -Streptavidin-Cy3 (12.5nM), H-NS 60-90 -Streptavidin-DyLight488 (50nM), H-NS 60-90 -Streptavidin-DyLight550 (12.5nM), H-NS 60-90 -Streptavidin-CF555 (12.5nM) or H-NS 60-90 -Streptavidin-CF488A (50nM) buffer C to bind fluorescent protein with amosite. After that, 20 μl of buffer C was added dropwise 3 times to remove unbound fluorescent protein. Finally, add 20 μl of water three times to remove the surfactant or salt from the buffer.

[0196] Next, place the filter collection surface on a glass slide (Mitsunami, MICRO SLIDE GLASS, 1 m...

Embodiment 3

[0202] [Example 3: Asbestos Microscopic Observation by Phase Contrast Fluorescence Method (2)]

[0203] In addition to the use of membrane filters that capture amosite (thea asbestos) (JAWE231) and rock wool (JFM standard fiber samples) as the test objects, and only use "H-NS 60-90 -Streptavidin-Cy3" was used as a fluorescent protein, and a specimen was prepared by the same method as in Example 2.

[0204] Observe the obtained specimen with a phase contrast / fluorescence microscope (Epi-fluorescence microscope BX-60, manufactured by Olympus). First, observe with a phase contrast microscope to confirm the fibrous material, then switch the optical path to fluorescence mode and observe the same field of view.

[0205] figure 2 It is a diagram showing the result of microscopic observation of asbestos by phase contrast fluorescence method. figure 2 (A) represents the phase contrast microscope image, (B) represents the fluorescence microscope image in the same field as the phase contrast ...

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Abstract

In order to provide a method capable of detecting asbestos more efficiently, easily, and with higher precision than phase-contrast microscope and electron microscope methods, without changing the criteria for detecting asbestos used in phase-contrast microscope and electron microscope methods, an asbestos-binding protein having a fluorescent marker is brought into contact with a substance to be detected, and a phase-contrast microscope and fluorescent microscope are then used in combination, whereby asbestos contained in the substance is detected.

Description

Technical field [0001] The invention relates to a method for detecting asbestos. Background technique [0002] Recently, the adverse effects of asbestos (fibrous silicate, also called "asbestos") on the human body are becoming a problem. That is, the company announced that people engaged in the manufacturing or use of asbestos in the past often suffered health damages such as lung cancer and mesothelioma. According to reports, through inhalation of asbestos dust, asbestos lung, lung cancer, malignant mesothelioma and other health problems are likely to occur. obstacle. [0003] Asbestos lung is one of the diseases of pulmonary fibrosis (pneumoconiosis) in which lung fibrosis occurs. There are many causes of pulmonary fibrosis such as other mineral dust, but pulmonary fibrosis caused by exposure to asbestos is particularly distinguished as asbestos lung. Lung cancer caused by asbestos fiber is caused by asbestos fiber taken into the alveoli mainly through physical stimulation. T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N21/27G01N21/78
CPCG01N2021/6439G01N33/5308G01N21/6428G01N21/6458
Inventor 黑田章夫石田丈典西村智基
Owner HIROSHIMA UNIVERSITY
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