Preparation of the Biological Safety Cabinet (bsc) for Cell Culture Work

SOP:        CC 6.6

Revision:         9.0

Title:        Preparation of the Biological Safety Cabinet (BSC) for Cell Culture Work

1.0 Purpose:

This procedure describes the proper method for preparing the Biological Safety Cabinet (BSC; a.k.a. Class II Laminar Flow Hood) prior to working with cell cultures and at the conclusion of the work.

2.0 Scope

Each time any type of work is to be done in the BSC the cleaning procedure described in this SOP must be followed before beginning work and at the conclusion of the work.  The BSC should also be cleaned when more than one cell line is to be used in a single day in the same BSC.  After completing work with one cell line the user must clean the BSC before introducing the new cell line into the BSC.

3.0 Responsibility

All users of the BSC without exception must be familiar with and understand this protocol. It is the responsibility of the lab manager / lab supervisor to verify and document that all users have been informed of this requirement and properly trained in the application of this protocol.  The lab supervisor / lab manager shall monitor adherence to this policy and take action as necessary to ensure compliance.

4.0  List of Abbreviations

4.1  BSC: Biological Safety Cabinet

4.2  IPA : isopropyl alcohol (2-propanol) or 70% ethanol

4.3  u.p.: ultrapure

4.4  U.V.: ultraviolet

5.0 Procedure

5.1 Materials

5.1.1        Biological Safety Cabinet (BSC)

5.1.2        Disinfectant Solution (L-pHast, Sporicidin, Wesocdyne, or similar agent) prepared at the proper concentration.

5.1.3        75% (v/v) Isopropyl alcohol (IPA)  

5.1.4        Wipe cloths - sterile, lint free (e.g. Stericloths or comparable)

5.1.5        Gloves

5.1.6        Sleeves (Optional)

5.1.7        Lab coat

5.1.8          Hair covering (Optional)

5.1.9        Safety glasses

5.2        Procedure for Routine (Daily) Cleaning of BSC

5.2.1        Be sure all items have been removed from BSC - it should be completely empty.

5.2.2          Lower sash to proper level (if applicable) and turn on the blower and U.V. light.  U.V. light is optional.  Allow air to circulate for at least 10-15 min prior to beginning work. Blower and U.V. light can remain on overnight. Do not work in room with U.V. light on.

5.2.3        Gown appropriately.  Ideally this will include hair cover, lab coat, gloves, closed toe shoes, sleeves, mouth cover, and safety glasses.

5.2.4        Turn off U.V. light and turn on white light while leaving blower on.

5.2.5        Obtain sterile solution of 75% isopropyl alcohol (IPA) or 70% ethanol in spray bottle and sterile lint free wipe(s).  Spray gloves, sleeves, IPA bottle and wipes with IPA solution.

5.2.6        Spray the interior of BSC with disinfectant solution (rotate the solution on a weekly basis) and wipe the BSC in the following order (you are wiping to evenly spread a thin layer of disinfectant over the entire surface and to dislodge any microbes -  not to wipe dry):

5.2.6.a        Left wall of BSC: wipe downwards with overlapping strokes from front to back.

5.2.6.b        Back wall of BSC: wipe downwards with overlapping strokes from left to right side - be sure to include the  corners.

5.2.6.c        Right wall of BSC: wipe downwards with overlapping strokes from back to front.

5.2.6.d        Floor of BSC: wipe from back to front in overlapping strokes starting from the left side and working to the right.

5.2.6.e        Interior of glass barrier:  wipe downwards with overlapping strokes from left to right side.