In-situ marine bacteria abundance and diversity detecting method based on laser induced fluorescence

Abstract

The invention discloses an in-situ marine bacteria abundance and diversity detecting method based on laser induced fluorescence. The method is wide in application range, quick and convenient to analyze and environment-friendly. According to the technical scheme, the method is characterized by comprising the steps that (1) a to-be-detected water sample solution is conveyed by a pump; (2) the water sample solution flows through a filter column; (3) a water sample flowing through the filter column enters a fluorescent pond; (4) a high-frequency pulse excitation light source-deep ultraviolet laser diode pumps a solid laser to generate a light source with a continuous wave band, the light source irradiates the fluorescent pond, a spectra light splitting device, namely a grating, on the detection side of the fluorescent pond performs spatial light splitting, chromatic dispersion is performed and then a lambda1-lambda2 spectral band is formed; (5) a CCD (charge coupled device) photoelectric detector positioned at a detection window simultaneously acquires lambda1-lambda2 spectral data, and internal transformation and time sequence integration are performed to obtain a wavelength-light intensity two-dimensional spectrum; and (6) a data processing system calculates and analyzes the abundance and the diversity of marine bacteria in a marine bacteria fluorescent identification mode.

Description

technical field [0001] The invention belongs to the field of detection methods for in-situ abundance and diversity of marine bacteria, in particular to a detection method for in-situ abundance and diversity of marine bacteria based on laser-induced fluorescence, which has a wide range of applications, quick and easy analysis, and is environmentally friendly. This method uses the optical characteristics of the fluorescence spectrum produced by the laser-induced fluorescence process and the "characteristic spectrum" of various marine bacteria itself - that is, the fluorescence spectra produced by various marine bacteria in the laser-induced fluorescence process are compared in different bands, Combined with marine bacterial fluorescence recognition patterns, the abundance and diversity of marine bacteria can be analyzed. Background technique [0002] Marine bacteria are an important part of the marine ecosystem. The study of bacteria in the marine environment is not only one ...

AI Technical Summary

Problems solved by technology

Due to the need to collect and prepare samples for these technologies, strict requirements for sample injection, and the need to make fluorescent markers, it is difficult to apply to the detection of bacteria in the marine field

At the same time, fluorescence microscope technology and plate culture methods have proved that in the ocean environment, only a small amount of marine bacteria can form colonies through traditional fixed plate technology, and some bacteria are in a living non-culturable state, which cannot be fully reflected by conventional isolation and culture methods. The state of marine bacterial resources and ecological functions, it is difficult to fully understand the abundance and diversity of bacteria and their ecological significance and role in the marine carbon cycle

[0004] In addition, the method adopts the mode of on-site sampling and analysis in the laboratory, that is, the method of on-site and real-time measurement cannot be realized, and other interfering substances are easily introduced during the sample transportation and processing process, which affects the accuracy of the analysis.

The detection of marine bacterial abundance and diversity involves certain environmental conditions and complex processes, so it is impossible to guarantee that there will be no possibility of secondary pollution in this process. The incompatibility of the environment, coupled with the large difference between the laboratory environment and the marine environment, leads to a large difference between the measurement results and the actual situation, and the accuracy and reliability of the results are questioned

[0005] The above methods have the following defects in varying degrees: 1. It must be completed in the laboratory, and the application cannot be real-time on site, and the scope is limited

2. The analysis takes a long time, at least a few days

3. The analysis process is complicated, the conditions are harsh, the energy consumption is high, and the technical level of the experimenters is high.

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