RISK ASSESSMENT is a process of evaluating/determining a

potential hazard, likelihood of suffering, or any adverse effects.

Factors/category determined risks:

•Operator (experience, training, protective clothing)

•Equipment (age, suitability, electrical safety, containment, heat,
maintenance, disposal)
•Physical risks (intense heat and cold, fire)
•Chemicals (including gases and volatile liquid)
•Biohazards (pathogenic, scale, genetic manipulation, containment)
•Radioisotopes (emission, volatility, localization on ingestion,
disposal)
•Special circumstances (pregnancy, illness, allergy etc)
•Elements of procedure (scale, complex, duration, number of persons
involved, location)
GENERAL SAFETY IN A TISSUE CULTURE LABORATORY
 
•Operator (experience, training, protective clothing)
•Equipment (maintenance, electrical safety, mechanical reliable)
ie toxic fume or aerosol from the centrifuge and homogenizer.
Should be contained or placed in fume cupboard.
•Glassware and sharp items (broken glasses, sharp bin, syringe
needles into metal contained. Accidental inoculation can causes
risk of transplantation)
•Chemical toxicity (distribution of powder and aerosols by
laminar flow hoods, advise to use liquid-based detergent or table
ie hypochlorite disinfectant, dimethyl sulfoxide DMSO,
mutagens, carcinogen, cytotoxic drugs, uses gloves)
•Gases (CO2, O2, N2). Keep in pressurized cylinder and secured.
Leakage of the gases causes risk of asphyxiation
•Liquid nitrogen (-196C, risk with frostbite, asphyxiation, and
explosion. Uses thick glove
•Burns (handling autoclave, ovens, and hotplates, naked flames
from Bunsen burner)
RADIATION
3 types of radiation hazards:
 

1. Ingestion
Soluble compound could be splashed on the hands or aerosol
generated via pipetting or the use of syringe.

Triated nucleotide incorporated in into DNA causes radiolysis

within the DNA. Example radioisotope iodine will concentrate in
thyroid and causes local damage.

Precautions: work in class II hood, wear gloves. Monitor spillage,

thorough clean up.
 
 
 
2. Irradiation from labeled reagents
Example: 32P, 125I, I13I and 51Cr.
Protection: 2-mm-thick lead shield, storing isotope in a lead
pot, work on tray in class II, Perspex screens (5mm)

3. Irradiation from a high-energy source.

X-ray machine, 60Co, or ultraviolet (UV) in sterilizing
apparatus or stopping cell proliferation in feeder layer.

Causes burn to the skin and damage eyes

Precautions: located in specified area, wear barrier filer

goggles.
Disposal of radioactive waste;
In designated sinks
Record the amount and disposal site.
Decontaminated the re-used vessel in (biological
decontaminant) hypochlorite and (radioactive
decontaminant) Decon
BIOHAZARDS

A biological agent, such as a virus or a condition that

constitutes a threat to humans, especially in biological
research or experimentation.

Horizontal laminar flow hoods >assure the sterility of the

culture is protected

Vertical Laminar flow with air-curtain hood > to prevent the

exposure of the operator to aerosol. These are defined as
Class II microbiological safety cabinets.
Levels of biological containments

4 biological safety levels: BSL 1, BSL 2, BSL 3, BSL 4.

Criteria:
•access,
•cleaning, personal hygiene,
•airflow and ventilation,
•equipment,
•sharps, MScs,
•disinfection,
•storage and transfer, disposal,
•biosafety manual and training,
•accident and spills,
•validation of facilities.
Microbiological safety cabinets

3. Maximum protection from know pathogens (a sealed

pathogen cabinet with filtered air leaving and
entering via a pathogen trap filter; Microbiological
safety cabinet Class III)

5. Intermediate level of protection for potential

hazards (A vertical laminar flow with front protection
in form of air curtain and filtered exhaust;
Microbiological safety cabinet Class II)

3. Minimal protection (open bench)

Human biopsy materials
 
1. Classified pathogen vs unclassified pathogen.
Unclassified pathogen: recombinant technique (transfection,
retroviral infection, interspecies hybridization)
 
2. Adventitious agents in human or other primate biopsy
samples or cell lines or animal products such as serum( from
endemic countries).
 
Handling precautions: samples in double wrapped container,
enter the logbook on receipt, work in class II biohazard
hood, avoid using sharp instruments, tape and label
container, proper disposal)
Genetic manipulation is procedure involved in altering
genetic constitution of the cells or cell line by
transferring nucleic acid.
 
Disposal of biohazardous waste should be put in a bag
and autoclaved/immersed in hypochlorite ie Clorox at
300-2500ppm.
 
Fumigation of the microbiological safety cabinet is
usually carried out by formaldehyde or Hydrogen
peroxide overnight.