This unit of competency is applicable to technical assistants working in laboratories in the biomedical, environmental, biotechnology and education industry sectors.

Industry representatives have provided case studies to illustrate the practical application of this unit of competency and to show its relevance in a workplace setting. These are found at the end of this unit of competency under the section 'This competency in practice'.

MSL973004A

Perform aseptic techniques

ELEMENT

PERFORMANCE CRITERIA

1. Work safely according to the legal and regulatory framework

1.1. Ensure work practices and personal actions conform to regulations, codes, guidelines and enterprise quality assurance procedures

1.2. Identify hazards and enterprise controls associated with the sample, preparation methods, reagents and equipment

1.3. Select, fit and use personal protective clothing and safety equipment

1.4. Address hazards and incidents as they arise

1.5. Maintain a chain of custody, traceable to the worker, for all cells and tissues

2. Prepare and test cell and tissue culture media

2.1. Select and confirm media specifications and processes/methods

2.2. Prepare culture media to suit the application

2.3. Sterilise culture media and check for sterility

2.4. Perform quality control checks to ensure that culture media is fit for purpose

2.5. Store culture media in accordance with specifications

3. Prepare tissue or cell cultures

3.1. Select tissue/cell sample to optimise growth and prepare it for culture

3.2. Add specified growth agents and/or nutrients

3.3. Inoculate culture medium using aseptic techniques

4. Monitor tissue or cell culture

4.1. Incubate culture in specified conditions

4.2. Monitor growth of culture and record appearance and characteristics

4.3. Report presence or absence of contamination

4.4. Subculture the culture to continue the cell line

4.5. Dispose of biohazardous and other laboratory waste safely

5. Maintain records

5.1. Maintain records of batches of media and test data

5.2. Ensure records of tissue cultures are retrievable, legible and accurate

5.3. Ensure records conform to information management, records, quality system and legal requirements

Required skills

Required skills include:

working safely

satisfying all legal and regulatory requirements, including the use and care of biohazard cabinets

preparing, diluting and sterilising reagents and culture media that are fit for purpose

growing cell lines and tissue to specifications without contaminating the original sample and the environment

identifying expected cell types and recognising normal and abnormal cells using an inverted microscope

counting cells (total and viable)

monitoring cell growth and recognising problems such as contamination

maintaining chain of custody, traceable to the worker, of all cell lines, tissues, logs of work completed and procedures/methods used

Required knowledge

Required knowledge includes:

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 and removal of wastes

types and sources of contamination

purposes and mechanisms of staining

importance of strict aseptic techniques and 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

The Evidence Guide provides advice on assessment and must be read in conjunction with the performance criteria, required skills and knowledge, range statement and the Assessment Guidelines for the Training Package.

Overview of assessment

Critical aspects for assessment and evidence required to demonstrate competency in this unit

Assessors should ensure that candidates can:

work safely and satisfy all legal and regulatory requirements, including the use and care of biohazard cabinets

prepare, dilute and sterilise reagents and culture media that are fit for purpose

grow cell lines and tissue to specifications without contaminating the original sample and the environment

identify expected cell types and recognise normal and abnormal cells using an inverted microscope

count cells (total and viable)

monitor cell growth and recognises problems, such as contamination

maintain chain of custody, traceable to the worker, of all cell lines, tissues, logs of work completed and procedures/methods used.

Context of and specific resources for assessment

This unit of competency is to be assessed in the workplace or simulated workplace environment.

This unit of competency may be assessed with:

MSL933001A Maintain the laboratory/field workplace fit for purpose

MSL973003A Prepare culture media

MSL973007A Perform microscopic examination.

Resources may include:

laboratory equipped with appropriate equipment, samples, cell lines and reagents

enterprise procedures and standard methods.

Method of assessment

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.

Where applicable, reasonable adjustment must be made to work environments and training situations to accommodate ethnicity, age, gender, demographics and disability.

Access must be provided to appropriate learning and/or assessment support when required.

The language, literacy and numeracy demands of assessment should not be greater than those required to undertake the unit of competency in a work like environment.

This competency in practice

Industry representatives have provided the case studies below to illustrate the practical application of this unit of competency and to show its relevance in a workplace setting.

Biotechnology

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.

Education

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 range statement relates to the unit of competency as a whole. It allows for different work environments and situations that may affect performance. Bold italicised wording, if used in the performance criteria, is detailed below. Essential operating conditions that may be present with training and assessment (depending on the work situation, needs of the candidate, accessibility of the item, and local industry and regional contexts) may also be included.

Codes of practice

Where reference is made to industry codes of practice, and/or Australian/international standards, it is expected the latest version will be used

Standards, codes, procedures and/or enterprise requirements

Standards, codes, procedures and/or enterprise requirements may include:

Australian and international standards, such as:

AS 1678 Emergency procedure guide -Transport

AS 2252 Biological safety cabinets

AS ISO 17025-2005 General requirements for the competence of testing and calibration laboratories

AS/NZS 2243 Set:2006 Safety in laboratories set

AS/NZS 2982.1:1997 Laboratory design and construction - General requirements

AS/NZS 4187:2003 Cleaning, disinfecting and sterilizing reusable medical and surgical instruments and equipment, and maintenance of associated environments in health care facilities

AS/NZS ISO 14000 Set:2005 Environmental management standards set

AS/NZS ISO 9000 Set:2008 Quality management systems set

HB 9-1994 Occupational personal protection

Australian code of good manufacturing practice for medicinal products (GMP)

Australian Dangerous Goods Code

client and product specifications

enterprise procedures, standard operating procedures (SOPs) and quality assurance procedures

gene technology regulations

Guide to physical containment levels and facility types

manufacturer's instructions or verbal direction from laboratory manager, supervisor or senior technician

material safety data sheets (MSDS)

National Code of Practice for the labelling of workplace substances [NOHSC:2012 (1994)]

occupational health and safety (OHS) national standards and codes of practice

operation and maintenance manuals for automated media preparation equipment

principles of good laboratory practice (GLP)

production schedules and instructions

Therapeutic Goods Regulations 1009

verified test methods

Applications of plant tissue/cell culture

Applications of plant tissue/cell culture may 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

Applications of animal tissue/cell culture may include:

establishment and maintenance of animal cell lines, such as liver, epidermal and 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

Hazards may include:

biohazards, such as infectious agents and oncogenic DNA

chemical and radiation hazards

allergenic factors

cryogenic liquids, such as nitrogen

heat from burners and molten agar

ultraviolet (UV) light

sharps, broken glassware

contaminated clothing

Hazard control measures and safety procedures

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, MSDS 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 and gowns

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

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 and pipette pumps

cell counting chambers and haemocytometer

incubators, including specialised atmosphere carbon dioxide

light and binocular inverted microscopes

centrifuges

Pre-use checks

Pre-use checks include:

performing routine maintenance

checks on raw materials and consumables, including use by date, possible contamination and storage conditions

Sterilisation and disposal of biohazardous wastes

Sterilisation and disposal of biohazardous wastes may include:

steam and high pressure air or steam

boiling, microwaving and autoclaving

filtration

gas, chemical and radiation

Plant tissues and cells

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 and somatic embryoids

tissue for callus development to initiate cell suspension cultures

Animal tissues and cells

Animal tissues and cells may include:

primary cells from animal tissue, such as heart, liver, kidney and epidermal

secondary cells, such as epithelial, endothelial and fibroblast

continuous cell lines, such as tumour lines, hybidomers and transformed lines (Epstein-Barr virus)

Preparing a primary culture

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

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

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 (e.g. Mycoplasma), fungi and other plant or animal tissue in the media

checking growth rates

performing viable cell counts

Subculture

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

Occupational health and safety (OHS) and environmental management requirements

OHS and environmental management requirements:

all operations must comply with enterprise OHS and environmental management requirements, which may be imposed through state/territory or federal legislation - these requirements must not be compromised at any time

all operations assume the potentially hazardous nature of samples and require standard precautions to be applied

where relevant, users should access and apply current industry understanding of infection control issued by the National Health and Medical Research Council (NHMRC) and State and Territory Departments of Health

Unit sector

Testing

This unit contains employability skills.