See Instructions in EMV´s HMS-page

Risk Assessment of
Hematoxylin/eosin staining of frozen sections
Department: EMV / Group: MuscleBiology / Date: 2013-04-09
Tasks included in the risk assessment
  • Filter hematoxylin
  • Prepareacidalcohol
  • Prepare Scott´s tap water
  • Filling/emptyingstainingjarswith hematoxylin and dip slides
  • Filling/emptyingstainingjarswithacidalcohol and dip slides
/
  • Filling/emptying staining jars with Scott´s tap water and dip slides
  • Filling/emptying staining jars with ethanol and dip slides
  • Filling/emptying staining jar with eosin and dip slides
  • Filling/emptying staining jars with xylene and dip slides
  • Mount slides in mounting medium
  • Clean and take care of waste
Sample preparation is not included in this risk assessment.

Includedchemicals, mixtures and solutions

Chemical, mixture or solution / CAS Number / Amount / Indication of danger(hazard statement andlimit value)
Hematoxylin / 517-28-2 / 200 ml / H302 Harmful if swallowed.
H315 Causes skin irritation.
H319 Causes serious eye irritation.
H335 May cause respiratory irritation.
Acidalcohol: HCl and 99,5% ethanol / 200 ml / H225 Highly flammable liquid and vapour
Level limit value / Ceiling limit value / Short-term value / Total dust / Respirable dust
[ppm] / [mg/m3] / [ppm] / [mg/m3] / [ppm] / [mg/m3] / [mg/m3] / [mg/m3]
500 / 1000 / 1000 / 1900
-Hydrochloricacid, conc37% / 7647-01-0 / 5 ml / H314 Causes severe skin burns and eye damage.
H335 May cause respiratory irritation.
Level limit value / Ceiling limit value / Short-term value / Total dust / Respirable dust
[ppm] / [mg/m3] / [ppm] / [mg/m3] / [ppm] / [mg/m3] / [mg/m3] / [mg/m3]
5 / 8
-Ethanol, 99,5% / 64-17-5 / 800 ml / H225 Highly flammable liquid and vapour
Level limit value / Ceiling limit value / Short-term value / Total dust / Respirable dust
[ppm] / [mg/m3] / [ppm] / [mg/m3] / [ppm] / [mg/m3] / [mg/m3] / [mg/m3]
500 / 1000 / 1000 / 1900
Scott´stapwater / 1000 ml / -
-Sodium bicarbonate, 2 g/l / 144-55-8 / 2 g / Not classified
-Magnesium sulphate, 10 g/l / 7487-88-9 / 10 g / Not classified
Ethanol, 80% / 64-17-5 / 200 ml / H225 Highly flammable liquid and vapour
Level limit value / Ceiling limit value / Short-term value / Total dust / Respirable dust
[ppm] / [mg/m3] / [ppm] / [mg/m3] / [ppm] / [mg/m3] / [mg/m3] / [mg/m3]
500 / 1000 / 1000 / 1900
Eosin Y, solution in ethanol / 15086-94-9 / 200 ml / Caution! – Not fully investigated!
Dissolved in flammable liquid
Ethanol, 95% / 64-17-5 / 700 ml / H225 Highly flammable liquid and vapour
Level limit value / Ceiling limit value / Short-term value / Total dust / Respirable dust
[ppm] / [mg/m3] / [ppm] / [mg/m3] / [ppm] / [mg/m3] / [mg/m3] / [mg/m3]
500 / 1000 / 1000 / 1900
Xylene, 100% / 1330-20-7 / 600 ml / H226 Flammable liquid and vapour.
H312 Harmful in contact with skin.
H315 Causes skin irritation.
H332 Harmful if inhaled.
Caution! Skin penetrating substance according to The Swedish Work Environment Authority’s provisions and on occupational exposure limit values, AFS 2011:18
Level limit value / Ceiling limit value / Short-term value / Total dust / Respirable dust
[ppm] / [mg/m3] / [ppm] / [mg/m3] / [ppm] / [mg/m3] / [mg/m3] / [mg/m3]
50 / 221 / 100 / 442
Pertex mounting medium, contains xylene and ethyl benzene / - / 1 drop / R10 Flammable.
R20/21 Harmful by inhalation, in contactwith skin and ifswallowed.
R38 Irritating to skin.

Identification of risks of accident or ill-health, assessment of risks and suggestion of preventive measures

Task / Identified risk/problem / Assessmentof risk, see risk rating / Preventivemeasures / Estimated risk after preventive measure
Consequence / Probability / Assessedrisk
Filter hematoxylin / Potential for exposure via inhalation and direct contact with eyes and skin. / Minor / Possible / High risk / Use labcoat, safety glasses and disposable nitrile gloves. Work in fume hood or on ventilated bench with Plexiglas hood.Write in handling and safety instruction. / Low risk
Prepare acid alcohol
/ Potential for exposure via inhalation and direct contact with eyes and skin.
Potential fire hazard. / Serious / Possible / High risk / Use labcoat, safety glasses and disposable gloves.
Prepare in fume hood. Write in handling and safety instruction. The limit values for hydrochloric acid and ethanol are not exceeded at any occasion.
All staff participates in mandatory fire training every 5 years.
Labs are equipped with fire extinguishers. / Low risk
Prepare Scott´s tap water / No risk associated with task / - / - / - / Use labcoat, safety glasses and disposable nitrile gloves. / -
Filling/emptying staining jars with hematoxylin and dipslides / Potential for exposure via inhalation and direct contact with eyes and skin. / Minor / Possible / High risk / Use labcoat, safety glasses and disposable nitrile gloves. Work on ventilated bench with Plexiglas hood. Write in handling and safety instruction. / Low risk
Dip slides in distilled water / No risk associated with task / - / - / - / Use labcoat, safety glasses and disposable nitrile gloves.
Filling/emptyingstainingjarswithacidalcohol and dip slides / Potential for exposure via inhalation and direct contact with eyes and skin.
Potential fire hazard. / Serious / Possible / High risk / Use labcoat, safety glasses and disposable nitrile gloves.
Work on ventilated bench with Plexiglas hood. Write in handling and safety instruction. The limit values for hydrochloric acid and ethanol are not being exceeded at any point.
All staff participates in mandatory fire training every 5 years.
Labs are equipped with fire extinguishers. / Low risk
Filling/emptyingstainingjarswithScott´s tap water and dip slides / No risk associated with task / - / - / - / Use labcoat, safety glasses and disposable nitrile gloves. / -
Filling/emptying staining jars with ethanol and dip slides / Potential for exposure via inhalation and direct contact with eyes and skin.
Potential fire hazard. / Serious / Possible / High risk / Use labcoat, safety glasses and disposable nitrile gloves.
Work on ventilated bench with Plexiglas hood. Write in handling and safety instruction. The limit value for ethanol is not being exceeded at any point.
All staff participates in mandatory fire training every 5 years.
Labs are equipped with fire extinguishers. / Low risk
Filling/emptying staining jar with eosin and dip slides / Potential for exposure via inhalation and direct contact with eyes and skin. / Minor / Possible / High risk / Use labcoat, safety glasses and disposable nitrile gloves.
Work on ventilated bench with Plexiglas hood. Write in handling and safety instruction.
All staff participates in mandatory fire training every 5 years.
Labs are equipped with fire extinguishers. / Low risk
Filling/emptying staining jars with xylene and dip slides / Potential for exposure via inhalation and direct contact with eyes and skin.
Potential fire hazard. / Serious / Likely / Veryhigh risk / Use labcoat, safety glasses and double, disposable nitrile gloves. Change gloves frequently, especially if you suspect contamination. Remove gloves carefully without touching skin. Avoid all contact with xylene.
Work in fume hood. Write in handling and safety instruction. The limit value for ethanol is not being exceeded at any point.
All staff participates in mandatory fire training every 5 years.
Labs are equipped with fire extinguishers. / Medium risk
Mount slides in mounting medium / Potential for exposure via inhalation and direct contact with eyes and skin.
Potential fire hazard. / Minor / Unlikely / High risk / Use labcoat, safety glasses and disposable nitrile gloves.
Work in fume hood. Write in handling and safety instruction.
All staff participates in mandatory fire training every 5 years.
Labs are equipped with fire extinguishers. / Low risk
Transport and handlechemicals and bottles / Dropbottles or chemicals potential risk for exposure via inhalation or directcontactwitheyes and skin.
Potential firehazard. / Serious / Possible / High risk / Store acids and basesbelowwaistheight. Transport on trolley or in a bucket. Make sure thatyouare prepared in caseof a spillage. Write in handling and safetyinstruction
Centralt placerade spillkit samt tillhörande instruktion tas fram, se EMV:s HMS-handlingsplan för 2013. / Low risk
Clean and take care of waste / Potential for exposure via direct contact with eyes and skin.
Potential environmental risk. / Serious / Possible / High risk / Same personal protective equipment as during the task
Send as chemical waste
Dispose of eosin and xylene as chemical waste. / Low risk
In case of power or ventilation failure or if evacuation is necessary / Fumesevaporatetolab / Serious / Possible / High risk / Close containers and close fume hood. Write in handling and safety instruction. / Low risk
Cleaners / If glass ware not rinsed before sending to be washed, potential for exposure via direct contact with eyes and skin. / Minor / Possible / High risk / Before sending glass ware to be washed it must be properly rinsed. Write in handling and safety instruction.
Cleaning must only be performed in free areas and cleaners must not wipe spill. Write ininstruction for cleaners. / Low risk
Have hazardous tasks been defined and rules for working alone been considered? / Yes
Have risks during power/water failure etc or because of evacuation been considered? / Yes
Do you have preparedness in case of spillage, eg first aid, absorption material? / Yes, in BMC Service
Do you have a written handling and safety instruction for this work? / Yes
Do you have a written instruction for cleaners and other maintenance personal? / Included in the Improvement plan for EMV 2013
References
  • Protocol for Hematoxylin/eosin stainingoffrozensections
  • SDS for all chemicals
  • LU´sHandling Regulations for Management ofhazardouswaste
  • EMV:s HMS-webpage

Participants
Katarina Johansson and Madeleine Durbeej-Hjalt
Reviewed by
Åsa Gustavson, Hans Hovenberg and Kristin Holmberg
Approved by Research Group Leader / Name in printing
Madeleine Durbeej-Hjalt