Chapter 2
Basic Information and Overview
1 Routine Precautions for Sterile Work
It is not necessary to spray alcohol in the cabinet before use. Turn the UV
light off and turn the fan and fluorescent light on. The gas burner cannot
be lit before the fan is operational in most laminar flow cabinets. While
working, keep the front glass shutter as low as possible, leaving only
enough space to insert your arms under the shutter.
It is recommended that you wipe your hands and arms with a 0.1 % aqueous solution of benzalkonium chloride prior to beginning work in the
laminar flow cabinet and that you repeat the wiping of your hands and
arms during the work. A 70% ethanol solution may be used instead of
benzalkonium; however, repeated wiping with ethanol may lead to rough
hands because alcohol extracts fat from your hands. All materials to be
used should be wiped with 0.1 % benzalkonium chloride before placing
them in the cabinet. When a glass medium bottle is opened, flame the
mouth and neck of the bottle for a few seconds. Use large forceps to hold
the cap of the bottle during the flaming process. If a screw-capped bottle
made of plastic is being used, wipe the neck of the bottle with 0.1 % benzalkonium chloride after removing the cap.
Pasteur pipette are used frequently. Pipettes that come in contact with
the cell suspension should not put back into the medium bottle. Doing
so will cause cellular contamination. To avoid this, it is recommended
that the pipette be changed each time it comes in contact with the cultured cells or the cell suspension. Or, if one cell line only is being handled,
clearly distinguish one pipette to be used for the culture medium and
another for cultured cells. Keep the tip of the pipette free from other
materials during the work. If the tip accidentally touches other materials, replace it immediately with a new one. Pasteur pipettes are used frequently with rubber caps or nipples. The rubber cap should be kept in
70% ethanol when not in use, because fungi may grow inside the cap,
especially if the inside of the cap has been wet with culture media.
2 Culture Media
Culture media is the most important factor affecting the results of culture. Three types of culture media are used. The first contains natural
substances as the main components. The second type of culture medium
OVERVIEW
Laminar
flow
cabinet
Disinfection
Pasteur
pipettes
Basic Information and Overview
1 Routine Precautions for Sterile Work
It is not necessary to spray alcohol in the cabinet before use. Turn the UV
light off and turn the fan and fluorescent light on. The gas burner cannot
be lit before the fan is operational in most laminar flow cabinets. While
working, keep the front glass shutter as low as possible, leaving only
enough space to insert your arms under the shutter.
It is recommended that you wipe your hands and arms with a 0.1 % aqueous solution of benzalkonium chloride prior to beginning work in the
laminar flow cabinet and that you repeat the wiping of your hands and
arms during the work. A 70% ethanol solution may be used instead of
benzalkonium; however, repeated wiping with ethanol may lead to rough
hands because alcohol extracts fat from your hands. All materials to be
used should be wiped with 0.1 % benzalkonium chloride before placing
them in the cabinet. When a glass medium bottle is opened, flame the
mouth and neck of the bottle for a few seconds. Use large forceps to hold
the cap of the bottle during the flaming process. If a screw-capped bottle
made of plastic is being used, wipe the neck of the bottle with 0.1 % benzalkonium chloride after removing the cap.
Pasteur pipette are used frequently. Pipettes that come in contact with
the cell suspension should not put back into the medium bottle. Doing
so will cause cellular contamination. To avoid this, it is recommended
that the pipette be changed each time it comes in contact with the cultured cells or the cell suspension. Or, if one cell line only is being handled,
clearly distinguish one pipette to be used for the culture medium and
another for cultured cells. Keep the tip of the pipette free from other
materials during the work. If the tip accidentally touches other materials, replace it immediately with a new one. Pasteur pipettes are used frequently with rubber caps or nipples. The rubber cap should be kept in
70% ethanol when not in use, because fungi may grow inside the cap,
especially if the inside of the cap has been wet with culture media.
2 Culture Media
Culture media is the most important factor affecting the results of culture. Three types of culture media are used. The first contains natural
substances as the main components. The second type of culture medium
OVERVIEW
Laminar
flow
cabinet
Disinfection
Pasteur
pipettes