Elements and Performance Criteria

The range of variables relates to the unit of competency as a whole. It allows for different work environments and situations that will affect performance.
Where reference is made to industry Codes of Practice, and/or Australian/international standards, it is expected the latest version will be used.
All operations must comply with legal/regulatory framework of regulations, codes and guidelines, relevant standards, appropriate procedures and/or enterprise requirements.

These procedures include or have been prepared from
Australian and international standards, such as -
AS ISO/IEC 17025 General requirements for the competence of testing and calibration laboratories
ISO 9000 series Quality management and quality assurance standards
AS 2243.2 Safety in Laboratories - Biological aspects
AS 2243.3 Safety in Laboratories - Microbiology
AS 2830 Good laboratory practice
AS 1678 Emergency procedures guide for hazardous materials
AS 2243.8 Fume hoods
AS 2252 Biological safety cabinets
AS 2982 Hand washing facilities
SAA HB9 Occupational personal protection, and other relevant standards for protective clothing (for example, AS 2161, AS 2210, AS 1337 and AS 1338)
AS 4187 Code of Practice for cleaning, disinfecting and sterilising reusable medical and surgical instruments and equipment, and maintenance of associated environments in health care facilities.
Office of the Gene Technology Regulator (OGTR) guidelines for working with genetically altered organisms
Good Laboratory Practice (GLP)
Therapeutic Goods Act
guidelines for the operation of classes of laboratories (for example, PCII, PCIII and PCIV physical containment)
enterprise procedures, standard operating procedures (SOPs), quality assurance procedures
verified test methods.

Applications of plant tissue/cell culture could include
mass propagation of commercial species
production of disease free plants by meristem tip culture
conservation of rare plants
haploid plant production by anther/pollen culture
'sports' produced by somaclonal variation
development of resistant plants by directed cell selection
protoplast fusion to produce novel plant hybrids.

Applications of animal tissue/cell culture could include
establishment and maintenance of animal cell lines, such as liver, epidermal, fibroblastic
maintenance of continuous cell lines
preparation of cell cultures for commercial sale
growth and enumeration of viruses
extraction of DNA
extraction of antigens for use in diagnostic tests
research of cell structure and function, cancer and tumour biology
immunofluorescent techniques
testing of media efficacy
production of monoclonal antibodies
production of genetically modified cell cultures
secondary metabolite production.

Hazards may include
biohazards, such as infectious agents, oncogenic DNA
chemical and radiation hazards
allergenic factors
cryogenic liquids, such as nitrogen
heat from burners, molten agar
ultraviolet light
sharps, broken glassware
contaminated clothing.

Hazard control measures and safety procedures may include
ensuring access to service shut-off points
recognising and observing hazard warnings and safety signs
labelling of samples, reagents, aliquoted samples and hazardous materials
handling and storage of hazardous materials and equipment in accordance with labelling, materials safety data sheets and manufacturer's instructions
identifying and reporting operating problems or equipment malfunctions
cleaning and decontaminating equipment and work areas regularly using enterprise procedures
using personal protective clothing and equipment, such as gloves, safety glasses, coveralls, gown
using containment facilities (PCII, PCIII and PCIV physical containment laboratories), containment equipment (biohazard containers, laminar flow cabinets, Class I, II and III biohazard cabinets) and containment procedures
following established manual handling procedures
reporting abnormal emissions, discharges and airborne contaminants, such as noise, light, solids, liquids, water/waste water, gases, smoke, vapour, fumes, odour and particulates to appropriate personnel.

Tissue culture equipment and facilities may include
growth cabinets
culture vessels, growth chambers, sterile containers, culture plates, flasks and bottles
autoclaves
positive filtration apparatus
auto pipettes, pipette pumps
cell counting chambers, haemocytometer
incubators, including specialised atmosphere carbon dioxide
light and binocular inverted microscopes
centrifuges.

Pre-use checks include
performing routine maintenance
checks on raw materials and consumables include use by date, possible contamination and storage conditions.

Sterilisation and disposal of biohazardous wastes may include
steam and high pressure air or steam
boiling, microwaving, autoclaving
filtration
gas, chemical, radiation.

Plant tissues and cells may include
plant tissue, such as petioles, leaves, stems and petals
meristem tissue
special tissue, such as fern stolon, seed embryos, somatic embryoids
tissue for callus development to initiate cell suspension cultures.

Animal tissues and cells may include
primary cells from animal tissue, such as heart, liver, kidney, epidermal
secondary cells, such as epithelial, endothelial, fibroblast
continuous cell lines, such as tumour lines, hybidomers, transformed lines (Epstein-Barr virus).

Preparing a primary culture may involve
thawing of cryopreserved cells and monitoring of cell recovery
enzymatic disaggregation from tissue
mechanical disaggregation from tissue
primary explant technique
pre treatment
disinfestation of explants using hypochlorite and water.

Suitable culture conditions may include
specified temperature and light intensity
appropriate atmosphere, such as carbon dioxide
shaking of cell suspensions or roller bottles
conditions for establishment, multiplication or planting out
special conditions for protoplast culture.

Monitoring growth of tissue and cell lines may include
identification of normal and abnormal cells viewed by an inverted stereo microscope
recognition of contamination, such as bacteria (eg Mycoplasma), fungi, other plant or animal tissue in the media
checking growth rates
performing viable cell counts.

Subculture may include
treatment of callus to multiply or regenerate shoots
treatment to encourage adventitious bud
treatment to encourage rooting
subculture of embryoids
cell suspensions
preparation of protoplasts.

Health, safety and environment
All operations to which this unit applies are subject to stringent health, safety and environmental (HSE) requirements, which may be imposed through State or Federal legislation, and these must not be compromised at any time. Where there is an apparent conflict between performance criteria and HSE requirements, the HSE requirements take precedence.
All operations assume the potential hazardous nature of samples and require standard precautions to be applied. Users should access and apply current industry understanding of infection control issued by the National Health and Medical Research Council and State and Territory Departments of Health. All operations are performed in accordance with standard operating procedures.

The Evidence Guide describes the underpinning knowledge and skills that must be demonstrated to prove competence.
Critical aspects of competency

Competency must be demonstrated in the ability to perform consistently at the required standard. In particular, assessors should look to see that the candidate:

works safely and satisfies all legal and regulatory requirements, including the use and care of biohazard cabinets
prepares, dilutes and sterilises reagents and culture media that are fit for purpose
grows cell lines and tissue to specifications without contaminating the original sample and the environment
identifies expected cell types and recognises normal and abnormal cells using an inverted microscope
counts cells (total, viable),
monitors cell growth and recognises problems, such as contamination
maintains chain of custody, traceable to the worker, of all cell lines, tissues and logs of work completed and procedures/methods used.

Underpinning knowledge

Competency includes the ability to apply and explain:

basic structure and function of cells and organelles
basic classes and classification of culturable material, such as organisms, plants, animals, bacteria, viruses, tissues, cells and prions
cell physiology and processes, such as simple and facilitated diffusion, plasmolysis, osmosis, tonicity, active transport, energy production, mitosis, motility, phagocytosis and pinocystosis
concepts and principles of cell growth, such as need for nutrients, role of growth regulators, removal of wastes
types and sources of contamination
purposes and mechanisms of staining
importance of strict aseptic techniques, cleaning procedures
hazards and risks in biological laboratories
relevant health, safety and environment requirements
enterprise and/or legal traceability requirements
relevant quality control checks and quality assurance procedures.

Assessment context and methods
This unit of competency is to be assessed in the workplace or simulated workplace environment.

The following assessment methods are suggested:

examination of tissue and cell cultures prepared by the candidate
observation of the candidate preparing a range of tissue and cell cultures
review of work records and results obtained by candidate
feedback from supervisors and peers on adherence to enterprise/technical procedures
questioning to assess underpinning knowledge.
In all cases, practical assessment should be supported by questions to assess underpinning knowledge and those aspects of competency which are difficult to assess directly. Questioning techniques should suit the language and literacy levels of the candidate.

Interdependent assessment of unit

This unit of competency may be assessed with:

PMLMAIN300B Maintain the laboratory fit for purpose
PMLTEST304B Prepare culture media
PMLTEST308B Perform microscopic examination.

Resource implications

Resources may include:

laboratory equipped with appropriate equipment, samples, cell lines and reagents
enterprise procedures, standard methods.

This competency in practice
A laboratory assistant maintains a leucocyte cell line which is used to routinely produce monoclonal antibodies which have been ordered by researchers. The assistant's job is to ensure that the cell line's growth is optimised to ensure a regular supply of high quality product. Every day, she/he checks for growth and contamination by aseptically removing a sample for microscopic examination. She/he also checks the colour of the pH indicator in the media and records cell line characteristics, such as its appearance, number of cells and any evidence of contamination in her/his laboratory notebook. She/he also checks the incubator temperature and atmosphere together with the labelling and possible leakage of flasks.
A laboratory assistant at a regional university is instructed to prepare 95 flasks of Vero (African green monkey kidney) cells for a practical class in three weeks time. She/he routinely passages the cells once per week and usually splits the flasks into six. She/he has three flasks routinely subcultured from last week and calculates that these can be subcultured to produce the required number of flasks while holding back some flasks from each subculture as a back up in case of contamination and for routine passaging after the practical class. She/he prepares the 95 flasks in the third week and checks them for obvious bacterial or fungal contamination and for Mycoplasma contamination. She/he labels all the flasks with the required information, records all the steps in the laboratory cell culture log and puts the flasks out in the teaching laboratory just prior to the class.

The seven key competencies represent generic skills considered for effective work participation. The bracketed numbering against each of the key competencies indicates the performance level required in this unit. These are stand-alone levels and do not correspond to levels in the Australian Qualifications Framework (AQF).
Level (1) represents the competence to undertake tasks effectively
Level (2) represents the competence to manage tasks
Level (3) represents the competence to use concepts for evaluating and reshaping tasks.
Communicating ideas and information
(1)

Collecting analysing and organising information
(2)

Planning and organising activities
(2)

Working with others and in teams
(1)

Using mathematical ideas and techniques
(1)

Solving problems
(2)

Using technology
(2)