Physiology and Cultivation of
Algae and Cyanobacteria
5.
Overview
• Collection of the samples
• Isolation & purification of algal culture
• Culturing
– Methods
– Equipments & material
– Conditions
– Culturing media
Collection of the samples
• purpose specific
• sample specifics (nature & environment)
• time
• concentration
• type of vessel/container
• removal of the unwanted organisms (filter)
• transfer conditions & storing
Isolation & purification of algal
culture
• Equipments & suplies
– Microscopes
– Filters & sieves
– Glassware, Plasticware, Utensils
• Methods
– Sterile manipulation
– Isolation techniques
Equipments & suplies
• Microscopes
– dissecting (80x,..)
– inverted
– lighting
• dark-field
• fiber optic light source
• fluorescent lamp
• Epifluorescence
• LED
• Filters & sieves
– woven screens
– nylon netting
– membrane filters (material)
– differential filtration
• Flow-box
Glassware, Plasticware, Utensils
• borosicate glassware
• plasticware (ready-to-use, culture-grade)
• sterilization technique & sterility
• dust-proof cabinet / clean containers
• caps & splips
• sterile filtration apparatus
• sterile spatula
• pens, labels
• parafilm
• growth chambers
– test tubes, flask, culture flask, Erlenmeyer flask, Petri dish, plugs, two-steps screw
– cultivator, tank, bioreactor
Sterilization & sterile manipulation
Methods of isolation & purification
• Enrichment culture
• Single-cell isolation
• Size separation >> filtering
• Density separation >> centrifugation
• Dilution
• Isolation with use of agar
– streaking
– spray
• Isolation with use of phototaxis
• Automatization (flowcytometer)
Culturing techniques
• chemicals
• equipment – balances, ph-meter, autoclave, filtration,
ultrasonic washer, refrigerator, cultivator
• conditions – ph, temp, irradiation, photoperiode,..
• glasware – E. flask, reagent bottles, pipetes, flask, tubes, P.
dishes, spatulas, funels, filter holder, syringe, ..
• water
• agar
• soil
Media
• stock solutions
– macronutrients
– trace elements
– vitamins
– chelators
– soil extracts
• preparation of media
• synthetic media
• enriched media
• soil water media
• solidified media – agar
• Freshwater media
• Seawater media
• natural & artificial