A quick way to test the effect of nanoparticles on algae activity

Abstract

The invention relates to a method capable of rapidly and accurately testing the activity of algae cells under the action of nanoparticles. The steps are: preparation of algae liquid culture and configuration of experimental solution; algae growth inhibition or inactivation test. During the test period, 100-400uL of the algae liquid after the test is dropped into the sterilized cell culture plate after a fixed period of time, and taken. Parallel sample; add cell proliferation reagent of 1 / 10 volume of algae liquid to the orifice plate algae liquid, incubate in the dark for 4-12 hours to make the cells adhere to the wall; absorb the supernatant and measure the absorbance at 450nm with a microplate reader or a spectrophotometer (OD) value. This method uses the absorbance of soluble formazan to measure the activity of mitochondrial enzymes in cells, which can indirectly reflect the activity of cell proliferation and avoid the interference of nanoparticles on counting. The method can be used to test the effect of nanoparticles in various water bodies such as sewage water, river water, various circulating water, and culture fluid on most algae including cyanophyta and green algae.

Description

technical field [0001] The invention relates to a detection method for the influence of nanoparticles on algae activity, which can quickly evaluate the inhibitory and killing effects of nanoparticles on algae, and facilitates the optimization of material properties. Background technique [0002] As a simple single-celled organism, algae has a high value in the fields of food, medicine and the environment, but algae outbreaks such as cyanobacteria blooms and red tides can also bring disasters to water ecology. Studies have proved that nanomaterials may be toxic to algae and play a positive role in the restoration of water body ecology, but the effect of nanomaterials on algae needs to be evaluated using a relatively simple and accurate method. Existing algae cell activity detection mainly refers to cell in vitro toxicity test standards, such as ISO1442:1999 and BS EN ISO8692:2004, but these standards do not clearly provide algae cytotoxicity test methods for nanomaterials, an...

AI Technical Summary

Problems solved by technology

Existing algae cell activity detection mainly refers to cell in vitro toxicity test standards, such as ISO1442:1999 and BS EN ISO8692:2004, but these standards do not clearly provide algae cytotoxicity test methods for nanomaterials, and all use cell concentration values As a cell viability standard, the effect of flocculation caused by nanoparticles is ignored on the results, and live cells cannot be distinguished from dead cells

Although some documents use fluorescent staining to count living and dead cells, they have high requirements for equipment and cannot quantify the activity of cells; the patent (CN101201312A) has invented a neutral red staining method, which does not need to be stained with fluorescent reagents. However, the interference of nanoparticles cannot be avoided, and the flocculation-sedimented algal cells still have the ability to proliferate; the chlorophyll a test method has a good linear correlation with cell activity, but when the cell activity negatively increases, its value has a poor correlation with chlorophyll content, and chlorophyll a It is easy to decompose under extreme stress conditions (strong light, strong acid and strong alkali, high temperature), so this method is only suitable for growth inhibition experiments under mild conditions; the cell proliferation reagent method is more accurate and can avoid the interference of various insoluble impurities, but For example, the MTT method is rarely used in the algae test. On the one hand, because the formazan produced by the MTT method is insoluble, the cells need to be lysed, and the steps are complicated. Tests are harmful to the experimental results and the health of the experimenters

Images