Prevention of Protocols’ hazards

Transformation Protocol

Hazards

1. Infection: The E.coli strain used is usually harmless but some of them can be harmful if not properly handled causing diarrhea abdominal pain.
2. Antibiotic Resistance:caused by plasmids which often carry genes that confer antibiotic resistance. If such plasmids are released into the environment, they can contribute to the spread of antibiotic-resistant bacteria.

Solutions

1. Wearing proper personal protective equipment, and sterilizing all equipment and materials.
2. Working in our AFCM lab of BSL-2 providing further infection control in addition to safety.
3. Proper training of lab personnel on handling the equipment( plasmid and E.coli) in addition to proper disposal of E. coli and plasmid DNA.
4. Avoid accidental ingestion or contact with open wounds in addition to proper antiseptic techniques.

Lipofectamine™ 3000 Reagent Protocol

Hazards

1. Lipofectamine reagent causes irritation upon contact with skin and eye.
2. Lipofectamine reagent can be harmful if it is swallowed or inhaled.

Solutions

1. Wearing proper personal protective equipment.
2. Using a biosafety cabinet to provide personnel protection.
3. Minimizing direct handling with the reagent through using pipettes.
4. Working in a well-ventilated area to make sure that airflow is proper to minimize the risk of inhalation.

Cell Culture Protocol

Hazards

1. Contamination of the culture by unwanted organisms.
2. Infection through inhalation of the aerosols.
3. Injuries during culture such as burns by chemicals , falls and cuts by sharp objects.
4. Harmness if used chemicals are mishandled.
5. Exposure to UV radiation that emits from biosafety cabinet.

Solutions

1. Following proper aseptic techniques , and working in a sterile and well-ventilated work environment.
2. Wearing proper personal protective equipment.
3. Disposal the waste materials safely.
4. Using caution during handling pipettes, needles and other sharp objects.


(Figure ) Pipettes

5. Handling carefully with chemicals.

Western Blot Sample Preparation Protocol

Hazards

1. Contamination by microbes leads to inaccurate results.
2. LDS Buffer and RIPA Lysis BUffer contain components which are irritant to skin and eye.
3. Halt Protease and Phosphatase inhibitor cocktail components are harmful , if inhaled or swallowed.
4. Injuries during culture such as burns by chemicals , falls and cuts by sharp objects.

Solutions

We followed aseptic techniques to prevent contamination of samples and reagents:

1. Wearing proper personal protective equipment.
2. Avoidance direct handling , inhaling or ingestion of the used chemicals.
3. Proper storage for the BCA reagent in a dry , cool place away from light sources.
4. Regular documentation of the records of western blot procedure and safety measurements.
5. Using caution during handling pipettes, needles and other sharp objects .

Gel Electrophoresis

Hazards

1. Ethidium Bromide is a known mutagen and acutely toxic irritant.
2. Acrylamide is a presumed carcinogen , a potent neurotoxin in its unpolymerized state and irritant upon contact.
3. Phenol is suspected carcinogen and toxic.
4. Usage of electrical currents in solutions is a potential risk for lab staff which may cause injuries.
5. Exposure to thermal hazards as burns by liquified gels.
6. Exposure to the UV radiation when visualizing the ethidium bromide gels.

Solutions

Wearing proper personal protective equipment.

1. Complete polymerization of acrylamide to minimize exposure to the unpolymerized form.
2. Using laboratory Tongs to avoid thermal hazards as burns during handling with samples.
3. Ensure electrophoresis is grounded to reduce the electrical risk.
4. Using UV-blocking glasses , and a timer to limit exposure to UV rays.

Flow Cytometry

(Figure) Flow Cytometry

Hazards

1. Infection through the pathogens which are in samples.
2. Contamination by microbes leads to inaccurate results.
3. Reagents such as EDTA (ethylene diamine tetraacetic acid) and staining buffers can be harmful if mishandled.
4. Fluorescent dyes which are used can be toxic.
5. The laser radiation the emits from cytometry can cause irritation to the eye.

Solutions

1. Follow aseptic techniques to prevent contamination of samples and used reagents.
2. Wearing proper personal protective equipment.
3. Proper handling with chemicals and avoiding direct contact.
4. Minimizing the exposure to the fluorescent dyes and laser radiation.

MTT Cell Viability Assay

Hazards

MTT cell viability assay involves several chemicals where each one of them causes a certain hazard:

1. HCL:Highly corrosive causing severe burn and damage to any tissue it touches like skin, eyes, respiratory tract and esophagus.
2. MTT:Photosensitivity and allergic reactions in addition to causing diarrhea and vomiting if ingested also some papers report having carcinogenic potential.
3. Proper handling with chemicals and avoiding direct contact.
4. SDS (Sodium Dodecyl Sulfate):Redness of the eye and damage of the GI tract causing nausea and vomiting diarrhea.

Solutions

1. Preventing any chemical ingestion or contact with open wounds in addition to the usage of proper antiseptic techniques which also prevents contamination of samples and used reagents.
2. Wearing proper personal protective equipment.
3. Proper ventilation in addition to proper storage of chemicals.

Three Color Stains in HEK293 cells

Hazards

1. Formaldehyde:According to International Agency for Research on Cancer (IARC) ,it is classified as a human carcinogen which has been linked to various cancers ( Nasopharyngeal cancer and also leukemia) moreover it can cause allergic reactions when applied to skin in some people (allergic dermatitis).
2. DAPI (4',6-Diamidino-2-phenylindole),ProLong Gold Antifade Reagent: Skin redness, eye irritation moreover if ingested or inhaled it can cause GUT discomfort and coughing ,respectively.
3. Triton X-100: According to GHS it can Acute oral toxicity, eye damage and irritation of the skin which can progress to corrosion.

Solutions

1. Preventing any chemical ingestion or contact with open wounds in addition to the usage of proper antiseptic techniques which also prevents contamination of samples and used reagents.
2. Wearing proper personal protective equipment.
3. Proper ventilation in addition to proper storage of chemicals.
4. Using a fume hood when working with Triton X-100 in addition to protective gloves and goggles.

References

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