TO ALL TEACHNING ON: Tissue Culture Course. BHS33-2 by I559m7B9


									                                                                                                . Neville A. Punchard. 2001

i). Basic cell culture laboratory skills, namely:
1.   Use, care and routine maintenance of basic tissue culture laboratory equipment, i.e.:
      Class II safety cabinet
      37oC CO2 Incubator
      Inverted phase contrast microscope
      Associated equipment, i.e. vacuum pump (if used), automatic pipette pumps, sterile plastic ware, etc.

2.   Basic sterile cell handling techniques and use of the Class II cabinet, i.e.:
      The structure of a cabinet, basic principles of use.
      Cleaning of cabinet before and after use, including use of UV light if applicable (don’t forget the bit about
          letting the cabinet airflow run for a while before and after use).
      How to set the cabinet up before use, i.e. work from sterile to non-sterile area, setting out bottles of reagents,
          beakers, waste containers, equipment, etc., to be used before starting work,
      How to remove flasks of cells from the incubator and into the cabinet to minimise risk of infection
      How to handle flasks of cells and sterile reagents in a cabinet while keeping them sterile,
      How to use sterile equipment in a cabinet.
      How to safely dispose of liquid and solid waste resulting from work in the cabinet and cell culture in general.
      What to do with contaminated glassware, etc.
      How to use other equipment in a cabinet, such as automatic pipette pumps and automatic pipettes
      How to examine a flask of cells using an inverted phase contrast microscope, including actual setting up of
          the microscope. What to look for in examining cells, such as: how to assess and report degree of confluency,
          and what it means; what floating cells in adherent culture can mean; what infections can look like, and
          generally how to assess their degree of success in cell culture.

3.   The "basic" use of the incubator i.e.:
      The structure and use of the incubator, i.e.: need for 37 oC, CO2 supply and water tray, monitoring all of these,
          how to look for sources of inflection in an incubator, and how you would clean an incubator, e.g.. if a user
          suspected it had a source of infection in it or for routine maintenance.
      How to set an incubator up from scratch
      How to monitor use of an incubator each time they use it, i.e. check temperature, water and CO 2 supply as
          first thing to do when entering the tissue culture lab and last thing before leaving it.
      How to place and remove flask from an incubator, e.g. loosening of caps and why needed, also different types
          of vented caps, sizes of flask available for use, etc.
      How to remove flasks of cells from the incubator and into the cabinet, and vice versa, to minimise risk of

4.   How to work independently in a tissue culture lab: by performing actual setting up and use of equipment and cell
     culture techniques on your own (after being taught/demonstrated the relevant techniques), with the need for
     minimal supervision.

5.   How to co-ordinate your work with that of other users in the use of the equipment and room, e.g. other
     staff/students using the cell culture laboratory.
6.   How to keep clean, and generally maintain, the cell culture equipment and laboratory.
7.   How to maintain continuous records of the cell cultures, i.e. using laboratory notebooks and central records


To top