A method for measuring the concentration of plant suspension culture cell liquid by micro sampling

Abstract

The invention discloses a method for trace detection of concentration of a plant suspension culture cell liquid, and the method includes the following steps: 1) acquisition of area data of stained cells in the cell liquid by trace detection, 2) acquisition of a standard curve of the area of the stained cells and the concentration of the cell liquid, and 3) establishment of a growth model of resistant Pinus densiflora 13-1 suspension subculture. The concentration of the cell liquid can be tested by trace detection, only 0.1mL of the cell liquid to be detected is sucked, under controlled sterile conditions, the cell liquid still can be cultured and proliferated; the method uses Image-ProPlus software to count, batch data processing time is saved, and artificial judgment errors can be reduced.

Description

technical field [0001] The invention belongs to the technical field of plant cell tissue culture, and in particular relates to a method for micro-sampling and measuring the concentration of plant suspension culture cell liquid. Background technique [0002] During the plant cell suspension culture process, the cells are evenly dispersed in the liquid medium, the nutrient utilization rate is high, and the proliferation speed is fast, and the suspension culture method can use large-scale bioreactors for large-scale industrial production of plant cells and their metabolites. [0003] The harvesting target of plant cell suspension culture is plant cells or their metabolites. However, plant cells can only maintain rapid growth within a certain concentration range, so it is of great significance to monitor the concentration of cell fluid in real time. Most of the plant cells are embryogenic stalk cell clusters, which are transparent, large, and easy to precipitate, so it is diffic...

AI Technical Summary

Problems solved by technology

[0004] At present, in the process of suspension primary culture, researchers mostly use cell sedimentation volume (SCV) to measure cell biomass and conduct quantitative research. After multiple cultures, plant cells are dispersed in the cell fluid, and the concentration of the cell fluid becomes thinner and less likely to settle; Therefore, in the face of the low concentration and high dispersion characteristics of suspension cell subculture, it is more difficult to quantitatively measure cell biomass.

At the same time, in the first generation of suspension experiments, it was found that the amount of cells needed for filtration and weighing and measuring SCV each time was too large, and the culture could no longer be continued after the measurement; when measuring the growth curve of cells for a long time, coupled with the design of experimental repetition, what needs to be done lots of experiments

Obviously, the SCV method to measure the concentration of cell fluid cannot fully meet the needs

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