Fluorescence spectrophotometry for detecting grease content of living algae cells in real time

Abstract

The invention discloses fluorescence spectrophotometry for detecting the grease content of living algae cells in real time and belongs to methods for rapidly screening the grease content of microalgae plants. According to the fluorescence spectrophotometry for detecting the grease content of the algae cells, microalgae of different species and different shapes is different in structure and shape, experiments and measurement are conducted on the microalgae, and dyeing conditions required by the microalgae of different species and different shapes are different, including selection of the concentration of solutions containing the algae cells, selection of dosage of fluorescence dye, selection of environments for the dyeing process and selection of parameters of fluorescence intensity measurement conditions; the concentration of the algae cells is 1*10<6> / ml-8*10<6> / ml; the volume of algae liquid is 5ml; the solubility of Nile red in the aqueous phase is 0.5-4 microgram / ml; vortex shaking time is 0.5-4 minutes. The fluorescence spectrophotometry has the advantages that complex steps that in traditional dry weight method grease measurement methods, it is required to make dry algae powder and conduct grease extraction and the like are omitted, algae liquid of the living cells is directly utilized for measuring the grease content in real time, rapidly and accurately, and microalgae plants high in grease content can be screened in a high-pass and rapid mode.

Description

technical field [0001] The invention relates to a rapid screening method for the oil content of microalgae strains, in particular to a fluorescence spectrophotometric method for real-time detection of the oil content of living algae cells. Background technique [0002] At present, among the traditional detection methods of algae cell oil content, the most classic experimental method is to extract algae oil by physical method and weigh and calculate, but this process is too cumbersome, which is easy to cause experimental errors and the shortcomings of inaccurate measurement accuracy. [0003] 1) Determination of oil content in microalgae cells, represented by BlighEG and DyerWJ, is the most classic method for oil determination in algae cells. The microalgae oil content determination method initially used the organic solvent extraction method to break the algae cell wall and extract the oil. This method can obtain more accurate data under good operating conditions, but the pro...

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Problems solved by technology

The microalgae oil content determination method initially used the organic solvent extraction method to break the algae cell wall and extract the oil. This method can obtain more accurate data under good operating conditions, but the process takes a long time and requires further analysis. Accurate weighing of algae cell raw materials, the choice of extractant and the removal degree of solvent also directly affect the reliability of data

[0004] 2) Nilered fluorescence spectrophotometry has been applied to the report of the determination of algae cell lipid content, but due to the water insolubility of Nilered reagent, as well as the pH of the environment and the ionic strength of the solution, the degree to which Nilered penetrates into algae cells is different, so the application This method must have a set of experimental standard parameters for detection, and the application of the unified experimental research method of Nilered fluorescence spectrophotometry for a certain microalgae has not been reported in detail

[0005] The above two methods have different disadvantages, resulting in the inability to measure the lipid content of living cells in a fast, real-time and high-throughput way:

[0006] (1) Determination of oil content by traditional dry weight method is cumbersome, such as the preparation of algae powder, oil extraction, oil drying, etc. It takes at least one week to complete, which cannot meet the requirements for rapid determination

[0007] (2) The existing Nilered fluorescence spectrophotometry is only a qualitative experiment, and there is no standard procedure for a certain microalgae experimental operation

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