Method for detecting whether nanoparticles exist in organ tissues or not
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A nanoparticle and tissue technology, which is applied in the direction of material analysis by measuring secondary emissions, can solve the problems of cumbersome nanoparticle methods and high cost, and achieve the effects of easy operation, reduced difficulty, and convenient search
Active Publication Date: 2014-12-10
HARBIN INST OF TECH
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[0003] The present invention aims to solve the problems of cumbersome and high-cost existing methods for detecting nanoparticles in organs and tissues, and provides a method for detecting whether there are nanoparticles in organs and tissues
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specific Embodiment approach 1
[0016] Specific embodiment one: the method for detecting whether there are nanoparticles in the organ tissue in this embodiment is carried out according to the following steps:
[0017] 1. Wash the organ tissue with PBS buffer, take 0.3-0.5g into a centrifuge tube, add 500μL RIPA lysate, and break;
[0018] 2. Add 500 μL RIPA lysate to the centrifuge tube, shake and mix well to obtain tissue fluid, and filter the tissue fluid with a filter to obtain filtrate;
[0019] 3. Take the filtrate and place it in a new centrifuge tube, extract it with saturated phenol for 1-3 times, discard the phenol layer, and obtain liquid A;
[0020] 4. Centrifuge liquid A at 14,000 g for 30 minutes, discard the supernatant, and obtain a precipitate;
[0021] 5. Suspend the precipitate with 100-200 μL alcohol, drop it on the surface of the aluminum plate or copper plate, and perform scanning electron microscope detection after the alcohol evaporates;
[0022] 6. After the nanoparticles are observ...
specific Embodiment approach 2
[0024] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the organs and tissues in Step 1 are fresh or frozen. Others are the same as in the first embodiment.
specific Embodiment approach 3
[0025] Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the crushing method in step 1 is ultrasonic crushing or mechanical stirring crushing. Others are the same as in the first or second embodiment.
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Abstract
The invention relates to a method for detecting whether nanoparticles exist in organ tissues or not, and solves the problems of complicated process and high cost of the conventional method for detecting nanoparticles in organ tissues. The method comprises the following steps: 1, cleaning the organ tissues, putting the cleaned organ tissues into a centrifugal pipe, and adding lysis liquid for breaking; 2, adding lysis liquid, uniformly shaking to obtain tissue liquid, and filtering the tissue liquid by a filter net to obtain filter liquor; 3, putting the filter liquor into a new centrifugal pipe, and extracting by saturated phenol to remove a phenol layer so as to obtain liquid A; 4, centrifuging the liquid A to obtain settlings; 5, suspending the settlings by alcohol, dropwise adding the suspension liquid onto the surface of an aluminum or copper plate, and carrying out detecting with a scanning electron microscope; 6, performing energy spectrum analysis to verify whether the nanoparticles are nanoparticles to be detected or not. According to the method, the fixing, embedding and slicing processes are eliminated, a sample preparation method is relatively simple and convenient, the operation is easy, and expensive slicing equipment is not required; the method is used for detecting whether the nanoparticles exist in the organ tissues or not.
Description
technical field [0001] The invention relates to a method for detecting nanoparticles. Background technique [0002] With the development of nanotechnology, nanomaterials have been applied in various fields closely related to human beings, such as pharmaceuticals, food additives, papermaking, sewage treatment, etc. However, a large number of studies have shown that eating or contacting these nanomaterials for a long time will have negative Therefore, nano-toxicity has become a key topic today. At present, the main point of view on the mechanism of the direct harm of nanomaterials to the body is that when nanomaterials enter the organs in the form of particles, their nanoscale volume and specific surface area will cause a large accumulation of reactive oxygen species in the cells of the organs, resulting in reactive oxygen species. Oxygen metabolism disorder eventually leads to oxidative stress, and oxidative stress is an important threat to cells, which can cause DNA damage,...
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