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Detection and analysis method for cell size distribution of phytoplankton

A technology of phytoplankton and analysis method, which is applied in the field of quickly obtaining the particle size distribution in the statistical sense, can solve the problems of lack of detection and analysis methods, unable to obtain the particle size distribution spectrum of phytoplankton cells, etc., and achieves high classification efficiency and sample demand. The effect of small amount and avoiding interference

Inactive Publication Date: 2016-08-17
CHONGQING INST OF GREEN & INTELLIGENT TECH CHINESE ACADEMY OF SCI
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Problems solved by technology

Due to the lack of effective detection and analysis methods, flow cytometry has not yet been used to detect the particle size distribution of phytoplankton, and it is also impossible to obtain a statistically significant particle size distribution spectrum of phytoplankton cells in water samples

Method used

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  • Detection and analysis method for cell size distribution of phytoplankton
  • Detection and analysis method for cell size distribution of phytoplankton
  • Detection and analysis method for cell size distribution of phytoplankton

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Embodiment 1

[0032] Flow cytometry was first used in the field of biomedical research. When a single cell with a fluorescein marker or an autofluorescent pigment passes through the laser irradiation area, it is excited to generate scattered light and fluorescent signals, and the strength of the forward scattered light (FSC) signal is related to the particle size of the detected cells. The fluorescent signal can be used to distinguish phytoplankton from other impurities such as sediment.

[0033] All phytoplankton contain photosynthetic pigments, which can spontaneously produce fluorescence when excited by laser light, and different pigments have different fluorescence characteristics. If excited by a 488nm laser, chlorophyll a emits red fluorescence (PerCP channel), and phycoerythrin emits orange fluorescence (PE channel). Chlorophyll a is the most basic photosynthetic pigment of phytoplankton, and all phytoplankton contain chlorophyll a. Therefore, using the PerCP channel to detect and ...

Embodiment 2

[0053] Taking a sample of phytoplankton in the backwater area of ​​a certain reservoir as an example, the particle size distribution of phytoplankton in the freshwater sample is detected and analyzed according to the method of the present invention. The operation process and results are as follows:

[0054] 1) Due to the collected fresh water sample concentration and the experimental requirement of 1×10 5 ~100×10 5 The cells / ml are consistent, so the sample is directly tested on the machine after passing through a 50μm sieve.

[0055] 2) In this example, a FACS Verse flow cytometer produced by BD Company was used for the experiment. Take 1ml of the pretreated sample, put it in the flow cytometer, adjust the test conditions and fluorescence compensation according to the method described in the patent, set the total number of stored cells to 10,000, and collect the experimental data. In the PerCP-FSC two-dimensional scatter diagram, the cells containing chlorophyll a (PerCP p...

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Abstract

The invention discloses a method for detecting and analyzing the particle size distribution of phytoplankton cells. 1) Pretreatment is performed on samples containing phytoplankton cells, and the cell concentration is adjusted to a suitable range; 2) According to chlorophyll a autofluorescence, the phytoplankton cells were screened out by cytometer analysis; 3) Under the same test conditions, fluorescent microspheres with different particle sizes were used to fit the Φ-FSC standard curve of forward scatter signal intensity (FSC) and particle size value Φ , to obtain a fitting function that converts the FSC signal intensity into a particle size value; 4) On the basis of step 2), generate a peak diagram of the particle number distribution of the phytoplankton cells injected after screening and the FSC distribution, and on the basis of step 3), Calculate the particle size corresponding to the mean value of the cell count in the injected sample to obtain the particle size distribution of the phytoplankton cells. The invention is easy to operate, can obtain the particle size distribution of phytoplankton in a statistical sense, and can be used to carry out research on particle size composition, ecological structure and the like of phytoplankton populations.

Description

technical field [0001] The invention belongs to the field of ecological environment research, and in particular relates to the use of flow cytometry to detect and analyze the particle size distribution of phytoplankton cells, in particular to quickly obtain the particle size distribution in a statistical sense. Background technique [0002] Phytoplankton is a general term for tiny plants that live in plankton in water. They are widely distributed in oceans and freshwater bodies around the world, and are the most important primary producers and energy converters in aquatic ecosystems. Among the many parameters studied on phytoplankton, particle size is a very important parameter. Phytoplankton metabolism, photosynthesis, energy utilization and other physiological characteristics are related to particle size. The contribution to productivity varies. [0003] In recent years, in the field of aquatic ecology and biogeochemistry, extensive research has been carried out on the pa...

Claims

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Application Information

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IPC IPC(8): G01N15/02
Inventor 李哲肖艳陶思圆郭劲松张振
Owner CHONGQING INST OF GREEN & INTELLIGENT TECH CHINESE ACADEMY OF SCI
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