STANDARD OPERATING PROCEDURE

HM550 Cryostat – HMRB 197

These guidelines are written to ensure that users of the HM550 cryostat in HMRB 197 are safe when
sectioning samples up to a classification of Biosafety Level 2 (BSL-2).

Authorization
Prior to start of work, ALL users must have:
• had an orientation by HBI AMP staff
• read this SOP
• submitted a completed HBI AMP orientation form
• undergone a detailed training with an experienced lab member
All users sectioning Risk Group (BSL-2) samples must have taken Biosafety training provided by
University of Calgary. Sectioning of human tissues or samples containing blood borne pathogen or
viruses must be reviewed to determine the risks involved to all users before use of the cryostat.

Hazards
SHARPS, Cold Temperatures (-20°C or below), Risk Group 2 Biohazardous Material

Personal protective equipment

Gloves & Lab Coats

Environment
When working in HMRB 197:
• No food or drink are allowed
• Close the door to limit access before starting your work
• Disinfect all working surface at the end of your work
• Wash your hands thoroughly at the end of your work
• Make sure you locate the First Aid Kit
• Be familiar with Emergency Procedures
• Ensure you have a personal cell phone with you at all times
• Use SoloSafe or Ensure a colleague knows if working alone or after-hours
• All incidents must be reported to HBI AMP staff and OARS
*DO NOT TOUCH DOOR HANDLES WITH GLOVES*

Emergency procedures
Follow the Post-exposure Protocol:
https://www.ucalgary.ca/risk/staff-wellness/occupational-health/post-exposure-protocol
*In case of serious emergencies contact 911 and campus Security 220-5333*

revised 9/17/2019
 STANDARD OPERATING PROCEDURE
HM550 Cryostat – HMRB 197

Using the cryostat in HMRB 197

1. Sign up for time on Faces Scheduling.

2. Bring your own consumables (brushes, blades & tweezer, OCT medium, slides) for preparation. All samples and
consumables must be carried to and from the cryostat (HMRB 197) in clean, closed, leak-proof,
impact resistant secure containers with absorbent material at the bottom of the containers.
3. PPE (gloves & lab coats) must be worn during handling and preparation of frozen sections. Do not
touch door handles with gloves! At any time, do not touch cold surfaces inside the cryostat for
extended periods of time to avoid cold burns.
4. Adjust temperature, let equilibrate, and mount samples.
5. Adjust section thickness and trim.
6. Place sample in holder and tighten clamp.
7. Insert blade into knife carrier (using tweezers), adjust position, and tighten both orange clamps.
8. WHEN NOT SECTIONING, WHEEL SHOULD BE LOCKED AND BLADE GUARD ENGAGED.
9. Make sure anti-roll plate and area adjacent to blade are clean.
10. Slowly cut tissue, brushing away from blade.
11. Stick sections on glass slides. TAKE EXTREME CARE OF SHARP BLADE.
12. At the end of your work
a. Lock the wheel
b. Remove blade (using tweezers) and discard in yellow bins
c. Reset temperature to -20°C
d. Remove all samples from the cryostat
e. Trimmings are considered BSL-2: remove ALL debris and discard in yellow bins
f. Disinfect hand-wheel and all top and inside surfaces with 70% ethanol
g. Close the window sash
h. Wash hands with soap and water
i. Log your time/issues in the log book by the entrance
13. All unused samples must be taken back to respective labs for disposal, transported as
described in #2 above.

revised 9/17/2019