In order to cultivate microalgae, it is first necessary to separate the desired "single species" or "clones" from the natural seawater if, in the course of cultivation, contamination with the algae or insect infestation occurs. The separation of the algae species must be re-introduced to ensure that the algae species have "single" or "growth" characteristics.
Based on their own experience and habits, microalgae growers have created a variety of methods for the separation and screening of algae species, which are more than a dozen. The most commonly used methods are the micropipette method, the water droplet method, and the agar plate method.
Micropipette method is the most commonly used, and it is one of the most reliable methods to obtain "single" or "clone". However, with high technical requirements, under the microscope, first use an anatomical needle made of pig's eyebrows to turn the debris in the water droplets and unwanted algal cells to the side and transfer the desired algae cells to the other side; The phototaxis of dinoflagellates can be applied to light on one side. The dinoflagellate to be selected is guided to the light side and the impurities remain in the original place. This makes it easier to suck the desired algal cells with a glass micropipette, and in addition, it is relatively large for individuals. The type can be operated under a double-dissecting microscope, and the algae cells in the micropipette are gently blown into shallow water slides or droplets on the slide glass. In order to achieve pure culture, the microalgae cells are preferably washed several times. , that is, after blowing in another droplet, sucking out another droplet of sterilized broth, repeatedly weighing it several times, and finally confirming by microscopic examination that the dripping is one algal cell to be screened, and then inoculating the test tube, or A section of capillary tube containing algal cells was pinched with sterile tweezers and placed in an inoculation tube.
The water droplet method is more convenient and convenient. First, a 24 by 24 mm cover glass was cut into small pieces of 8 by 8 mm with a small grinding wheel of an ampoule, washed and dried, and placed in a Petri dish. Bacteria, arrange these small slides on a sterilized glass slide, and in each small slide, drop a small drop of water containing microalgae, the size of the water droplets to see the water droplets under the microscope at low magnification. All or most of the time, the slightest movement of the small water droplets can be seen by moving the microscope stage slightly. It is found that only one small algae cell to be separated from the small water droplets can be used to disinfect the glass when other organisms are mixed. The tablets were sandwiched and placed in a small test tube containing culture medium for cultivation. If it is not to obtain "clones", as long as they are the same type of algae cells in the water droplets, 2-3 cells will form a chain, which will make it easier for the algae needed for rapid growth and reproduction.
The agar plate method is most suitable for the separation of energy-moving pinnate diatoms and flagellar green algae, and the method is easy to grasp. This can be divided into streak inoculation, spray inoculation and water droplet diffusion. Now only take the droplet diffusion method as an example. In the prepared 1-1.5% agar, heat and stir, sub-packed in petri dishes, thick 3-5 mm, autoclaved, let cool), sparsely dripped some water droplets, put Incubation into the incubator or under the northern window, when the algae in the alfalfa reservoir diffuse with the water droplets and the movement of the algae itself, disperse into a single individual, form algae on a certain part of the plate in some aspects, check and confirm under the microscope Afterwards, the required algae are removed with the sterilized glass micro-needles, put into a test tube, and repeated several times to obtain a pure-bred culture.
Based on their own experience and habits, microalgae growers have created a variety of methods for the separation and screening of algae species, which are more than a dozen. The most commonly used methods are the micropipette method, the water droplet method, and the agar plate method.
Micropipette method is the most commonly used, and it is one of the most reliable methods to obtain "single" or "clone". However, with high technical requirements, under the microscope, first use an anatomical needle made of pig's eyebrows to turn the debris in the water droplets and unwanted algal cells to the side and transfer the desired algae cells to the other side; The phototaxis of dinoflagellates can be applied to light on one side. The dinoflagellate to be selected is guided to the light side and the impurities remain in the original place. This makes it easier to suck the desired algal cells with a glass micropipette, and in addition, it is relatively large for individuals. The type can be operated under a double-dissecting microscope, and the algae cells in the micropipette are gently blown into shallow water slides or droplets on the slide glass. In order to achieve pure culture, the microalgae cells are preferably washed several times. , that is, after blowing in another droplet, sucking out another droplet of sterilized broth, repeatedly weighing it several times, and finally confirming by microscopic examination that the dripping is one algal cell to be screened, and then inoculating the test tube, or A section of capillary tube containing algal cells was pinched with sterile tweezers and placed in an inoculation tube.
The water droplet method is more convenient and convenient. First, a 24 by 24 mm cover glass was cut into small pieces of 8 by 8 mm with a small grinding wheel of an ampoule, washed and dried, and placed in a Petri dish. Bacteria, arrange these small slides on a sterilized glass slide, and in each small slide, drop a small drop of water containing microalgae, the size of the water droplets to see the water droplets under the microscope at low magnification. All or most of the time, the slightest movement of the small water droplets can be seen by moving the microscope stage slightly. It is found that only one small algae cell to be separated from the small water droplets can be used to disinfect the glass when other organisms are mixed. The tablets were sandwiched and placed in a small test tube containing culture medium for cultivation. If it is not to obtain "clones", as long as they are the same type of algae cells in the water droplets, 2-3 cells will form a chain, which will make it easier for the algae needed for rapid growth and reproduction.
The agar plate method is most suitable for the separation of energy-moving pinnate diatoms and flagellar green algae, and the method is easy to grasp. This can be divided into streak inoculation, spray inoculation and water droplet diffusion. Now only take the droplet diffusion method as an example. In the prepared 1-1.5% agar, heat and stir, sub-packed in petri dishes, thick 3-5 mm, autoclaved, let cool), sparsely dripped some water droplets, put Incubation into the incubator or under the northern window, when the algae in the alfalfa reservoir diffuse with the water droplets and the movement of the algae itself, disperse into a single individual, form algae on a certain part of the plate in some aspects, check and confirm under the microscope Afterwards, the required algae are removed with the sterilized glass micro-needles, put into a test tube, and repeated several times to obtain a pure-bred culture.