Lab sessions in Biotechnology

Cultivation of pure cultures, i.e. only one species of microorganism, is carried out in Erlenmeyer flasks in shaking water baths in liquid volumes of 50-150 mL, or in bioreactors with liquid volumes of 4 L, 20 L or 1000 L on the desired substrate.

Equipment

• Shaking water bath for Erlenmeyer flasks
• Bioreactor bubble column or airlift: 4 L liquid volume
• Bioreactor bubble column or airlift: 20 L liquid volume
• Bioreactor bubble column: up to 1000 L liquid volume 

Analysis

Primary analysis of substrates and metabolites, see also the section Analysis linked to biotechnological experiments under the lab module Cultivation of human and animal cells. 

Mixed microbial cultures, such as sludge for anaerobic digestion or rumen fluid from animals, are used for the production of biogas and volatile fatty acids which are further valorised into single cell protein for animal feed, biofuels such as butanol and hexanol, bacterial bioplastics such as polyhydroxyalkanoates (PHAs), carbon source for denitrification in wastewater, etc. The cultivation of mixed microbial cultures is carried out in bioreactors such as serum bottles, benchtop CSTRs, and specialised membrane bioreactors on a laboratory and pilot scale.

Equipment

• Anaerobic Reactors 120 ml – 31 L
• Membrane bioreactors 4 L – 500 L
• Sidestream micro, ultra and nanofiltration units 0.5 L – 50 L
• Trickle bed bioreactors for bioconversion of gases 10 L
• Incubators 35–55 degrees
• Anaerobic chamber/glovebox
• BPC High Precision Gas Volume and Flow Meter
• Furnaces and muffle furnaces
• Spectrophotometers
• Viscometer/Rheometer

Analysis

• GC Unit for CO₂/CO/H₂/CH₄ measurements
• GC Unit for Volatile Fatty Acid Analysis
• HPLC and LC-MS for the measurement of alcohols, acids, furans and sugars
• Elemental analyser for C/H/N/S measurement
  Kjeldahl unit for nitrogen determination
• Fibre analyser
• Fat analyser
• Atomic emission spectroscopy for quantification of metal elements

Culture of human and animal cell lines is carried out in T-flasks under controlled conditions in cell culture biosafety cabinets. In order to maintain a viable and long-term stable culture, the cells are regularly subcultured and can then be used in various types of experiments, such as cytotoxicity analyses.In order to maintain a viable and long-term stable culture, the cells are regularly subcultured and can then be used in various types of experiments, such as cytotoxicity analyses.

Equipment

• Class II biological safety cabinet
• CO₂ Incubator
• Inverted microscope for cell monitoring
• Microplate reader

Analysis 

Primarily, the morphology and viability of the cells are analysed. Microscopic images are taken to document cell state and the degree of confluence can be estimated. For functional tests, MTT analysis can be performed to measure cell viability after treatment, for example in cytotoxicity studies.

Pre-treatment of hard-to-degrade substrates is important to improve degradation and increase the efficiency of biological processes. By pre-treating the substrate – for example with mechanical, thermal, chemical or biological methods – structures are broken down, cell walls are opened and the availability of microorganisms increases. 

Equipment

• Parr reactor for chemical reactions at high temperature and pressure, such as biomass pretreatment
• Ultra-fine friction grinder “Supermasscolloider” for homogenisation and fibrillation of soft materials, e.g. Biomass
• Rotary evaporator with capacity of 2-20 litres, for concentrating solutions, biomass pre-treatment and cellulose purification
• Rotor bolt grinder for grinding down dry materials, such as wood chips and bread
• Liquid nitrogen generator, for e.g. freeze-thaw cycle with liquid nitrogen for pre-treatment of materials and extraction of various substances

Analysis linked to biotechnological experiments

• Oven for dry weight determination
• Ultrafiltration
• Centrifuges with different capacities for separating materials of varying densities
• Spectrophotometer
• FTIR for analysis of primarily pre-treatment and its impact on the substrate
• Analysis of polysaccharides, including cellulose, hemicellulose, chitin and chitosan, as well as analysis of lignin
• Analysis of how different materials affect cells through cell culture experiments, with a focus on cytotoxicity, biocompatibility, cell viability and proliferation
• Investigation of antibacterial properties of different materials.
• Texture analyser
• HPLC for the quantification of metabolites, organic acids, common inhibitors that can be formed in biomass pre-treatment, and simple sugars
• GC for Analysis of Gases and Fatty Acids (VFAs)
• Kjeldahl and Elemental analyser for determination of nitrogen (including crude protein), carbon, hydrogen and sulphur
• Microscope (including oCelloScope – automated imaging and analysis of living microbial cells)