Simple and efficient method for separating and purifying unicellular algae

Abstract

The invention discloses a simple and efficient unicellular algae separation and purification method, which comprises the following steps: accurately measuring the unicellular algae cell density of an algae solution to be separated and purified by using a blood cell counting plate under an optical microscope, diluting the algae solution to the unicellular algae cell density of 20-30 cells / mL by using sterilized sand filtration seawater, sucking 1-3 drops of diluted algae liquid according to the quantity of miscellaneous algae cells and harmful organisms in the algae liquid to be separated and purified, dripping the diluted algae liquid into a glass test tube filled with a unicellular algae culture solution, culturing for 10-15 days in a place with sufficient illumination and without direct sunlight, and carrying out microscopic examination and screening to obtain purified unicellular algae without miscellaneous algae cells and harmful organisms. The method disclosed by the invention is simple to operate, extremely low in requirement on professional degree of operators, wide in application place, suitable for laboratories with good conditions and farms with simple conditions, and capable of ensuring pure culture and long-term storage of the unicellular algae, and the success rate of separation and purification reaches 100%.

Description

technical field [0001] The invention relates to the field of unicellular algae cultivation, in particular to a simple and efficient method for separating and purifying unicellular algae. Background technique [0002] As bait, Unicellular algae has been widely used in the cultivation of economically cultured seedlings. However, during the process of seed preservation and inoculation of Unicellular algae, contamination by miscellaneous algae cells or harmful organisms often occurs, resulting in the abnormal growth and death of Unicellular algae. Therefore, it is necessary to separate and purify the algal species to ensure monoculture and normal growth and reproduction, so that it can be applied to scale-up productive cultivation. [0003] The traditional separation and purification methods of Unicellular algae are mainly water drop method and plate separation method. In the water drop method, use a capillary pipette to absorb the diluted algae liquid, drop 4 to 5 micro-water ...

AI Technical Summary

Problems solved by technology

In the water drop method, use a capillary pipette to absorb the diluted algae liquid, drop 4 to 5 micro-water droplets scattered on the slide, and then examine the unicellular algae cells, harmful organisms and miscellaneous algae cells in each micro-water droplet under a microscope. In the case of unicellular algae cells, no harmful organisms and miscellaneous algal cells, the micro-water droplets are sucked with a micropipette and transferred to a test tube containing a culture medium for cultivation, but the water-drop method has the following three disadvantages: (1) In order to ensure There is only one cell in each water droplet, and the algae solution must be properly diluted first, and the degree of dilution needs to be continuously adjusted according to the number of algae cells in the micro-water droplet, which is cumbersome to operate and takes a long time; The salinity rises sharply, causing the Unicellular algae cells to shrink and deform or even die, which reduces the success rate; (3) It cannot be guaranteed that the Unicellular algae cells in the micro-water droplets are accurately absorbed by the micropipette, and may still remain on the glass slide. reduced success rate

Based on this, the patent CN107384794A has further improved the water drop method. Each drop of water sample is dropped on a small-sized carrier sheet that can be directly put into the culture test tube. After being selected by microscopy, the selected carrier sheet is directly placed in the Cultivate in the culture test tube, the method of this patent makes up for the above-mentioned third point deficiency, but there are still first two points of deficiency

In addition, another method of separation and purification of Unicellular algae——in the plate separation method, use an inoculation loop to dip the algae liquid and gently streak it on the agar medium containing the nutrient salt of the Unicellular algae, so that the algal cells are serially diluted on the line. After the growth and reproduction of the single cell algae cell group into the single cell algae, it is placed in the test tube for culture, but the preparation process of the plate is cumbersome, and the water in the agar medium is easy to be separated and purified during the 10-15 day separation and purification process. Evaporate and dry out, resulting in shrinkage, deformation and even death of the cells of the unicellular algae

[0004] It can be seen that the success rate of the existing methods for separation and purification of Unicellular algae is extremely low, and it can only be carried out under good laboratory conditions, and it is difficult to carry out in farms with poor conditions. , the development of a single-celled algae separation and purification method with simple operation, high purification success rate, and wide application is quite necessary for the field of single-celled algae cultivation