Herzenberg Lab Human Blood Handling Guidelines

April 1999 Revision

Preliminary Information

1) All individuals working in the lab should have received a Hepatitis B vaccination. Call the Biosafety Office, 725-1473, for further information.

2) All individuals working with human samples should have completed the Stanford Bloodborne Pathogen training program on the internet:

http://somsafety.stanford.edu/bbp2/index.html

3) Contact Information:

Health & Safety Programs main office:

Phone: (650) 723-7233

E-mail: ma.gic@forsythe.stanford.edu

Buffy Coats / Whole Blood

1) Log in all buffies into the human blood samples binder located in B011 above Steve's bench.

2) Vacutainers containing blood should be stored/transported in orange capped biohazard containers.

3) Buffy coats should be treated as potentially infectious even after they are tested. They should always be handled in a manner as to minimize the risk of infection.

4) Label hoods, centrifuge and benches with a Processing Human Blood label containing the date and initials of the user.

5) Always wear a lab coat and gloves when working with human samples. Double gloving is recommended when handling blood.

6) Prepare a red autoclave bag to dispose of all contaminated waste. Use a red sharps container with a human samples label to dispose of contaminated pipettes and vacutainers.

7) Use a Fenwal transfer kit to extract buffies. Do not use scissors/sharps.

8) Use 20% bleach to decontaminate all containers/vacutainers prior to disposal.

9) Prepare a beaker with 20% bleach to decontaminate pipettes prior to disposal.

10) Remove gloves and wash hands when moving to other areas or handling common use equipment.

11) Immediately clean all spills with 20% bleach.

12) After completion, wipe off all surfaces with 20% bleach followed by dH2O and 70% ethanol. Flush aspirator with 20% bleach.

13) Seal filled sharps containers, place them in autoclave bags, mark the bags, "Biohazard", and place the bags on the cart in B013.

14) Remove warning labels from work areas after you are done.

Staining Human Samples

1) PBMC's and other cells derived from human blood should be treated as potentially infectious. After isolating from human blood, PBMC's can be brought to the bench top. Use extra care when handling them outside a hood.

2) Label work benches and centrifuges with a Processing Human Blood label containing the date and intials of the user.

3) Always wear a lab coat, gloves and eye protection when working with human samples.

5) Do not consume food or beverages in areas where human samples are being handled.

6) Dispose of all contaminated waste in a red autoclave bag. Use a red sharps container with a human samples label to dispose of contaminated pipettes and vacutainers.

7) Be sure to fix stained cells in 0.5% paraformaldehyde before running them on FACS.

8) After completion, wipe off all surfaces with 20% bleach followed by H2O and 70% ethanol.

9) Double bag autoclave bags, mark the bags, "Biohazard", and place bags on the cart in B013.

10) Seal filled sharps containers, place them in autoclave bags, mark the bags, "Biohazard", and place the bags on the cart in B013.