Great Bay Community College Document Number: 1.5.

3
320 Corporate Drive Revision Number: 3
Portsmouth, NH 03801 Effective Date: 18Jul08
Page 1 of 4
Title: Sanyo CO2 Incubator MCO-180IC Operation SOP
Approvals:
Preparer: _ Bob O‘Brien _______________________________Date: __17Jul08___________
Reviewer: _Deb Audino_______________________________ Date: __17Jul08___________

1. Purpose:
1.1. Operation of the Sanyo CO2 incubator.
2. Scope:
2.1. Applies to operation of the incubator for mammalian cell culture.
3. Responsibilities:
3.1. It is the responsibility of the course instructor/lab assistant to ensure that this SOP is
performed as directed and to update the procedure when necessary.
3.2. It is the responsibility of the students/technicians to follow the SOP as described and to
inform the instructor about any deviations or problems that may occur while performing
the procedure.
4. References:
4.1. Sanyo instruction manual MCO-18AIC.
5. Definitions: N/A
6. Precautions:
6.1. There is a possibility of oxygen depletion in the vicinity of the equipment.
6.2. Do not lean on the chamber doors.
7. Materials:
7.1. liquefied CO2 gas bone dry cylinder
7.2. CO2 gas regulator
7.3. tubing
7.4. tubing clamps
7.5. biopure water (filtered deionized water)
7.6. 70% isopropanol (IPA)
8. Procedure:
Note: Refer to Figures 1 and 2 as needed throughout this procedure.
8.1. Setup
8.1.1. Turn power on (if necessary).
8.1.2. The current incubator temperature and CO2 level will be displayed on the
control panel.
8.2. Set the temperature.
8.2.1. Press the Set key on the control panel.
8.2.2. The left most temperature digit will automatically flash.
8.2.3. Press the Numerical Value Shift key ▲ ▲ to change the numerical value to the
desired temperature.
8.2.4. Press the Digit Shift key ►► to shift digit positions to the middle position. The
temperature digit will automatically flash.
▲
8.2.5. Press the Numerical Value Shift key ▲ to change the numerical value to the
desired temperature.
8.2.6. Press the Digit Shift key ►► to shift digit positions to the right most position.
Great Bay Community College Document Number: 1.5.3
320 Corporate Drive Revision Number: 3
Portsmouth, NH 03801 Effective Date: 18Jul08
Page 2 of 4
Title: Sanyo CO2 Incubator MCO-180IC Operation SOP
8.2.7. Press the Numerical Value Shift key ▲ ▲
to change the numerical value to the
desired temperature.
8.2.8. Press the “ENT” key to allow the controller to accept the input temperature.
8.3. Set up the CO2 feed.
8.3.1. Verify that the tubing clamp is open to allow CO2 gas into the feed supply for
the incubator.
8.3.2. Verify that the open/close valve on the CO2 cylinder is in the open position and
that the CO2 regulator is turned on to the required pressure.
8.3.3. Press the “Set” key to set CO2.
8.3.4. The left most temperature digit will automatically flash, and the left most
carbon dioxide digit will automatically flash.
8.3.5. Press the Numerical Value shift key ▲ ▲
on the control panel until the display
shows the desired operating CO2 level.
8.3.6. Press the Digit Shift key ►►to shift digit positions and the Numerical Value
Shift key ▲
▲
to change the numerical value.
8.3.7. Press the enter key twice on the control panel to enter the percentage of CO2
into the memory.
8.3.8. When the CO2 density is set to 0.00, the control is off regardless of the chamber
density.
8.4. Operation
8.4.1. Verify that the actual temperature and percentage of CO2 are at the desired
levels.
8.4.2. Clean the outer surface of the vessel containing the culture or sample to be
incubated with 70% IPA prior to placing it in the incubator.
8.4.3. Open the outer incubator chamber door.
8.4.4. Open the inner incubator chamber door.
8.4.5. Place the culture to be incubated in the center of the tray if possible.
8.4.6. Close the inner incubator chamber door.
8.4.7. Close the outer incubator chamber door.
8.4.8. Verify that the operating temperature and CO2 levels on the control panel are set
to the recommended levels for the culture or sample being incubated.
9. Attachments:
9.1. Figure 1: Incubator Components
9.2. Figure 2: Control Panel
10. History:
Name Date Amendment
Bob O’Brien 03Feb05 Initial Release
Bob O’Brien 13Apr07 Updated the date format. Added key symbols to directions as
needed.
Bob O’Brien 18Jul08 College name change
Great Bay Community College Document Number: 1.5.3
320 Corporate Drive Revision Number: 3
Portsmouth, NH 03801 Effective Date: 18Jul08
Page 3 of 4
Title: Sanyo CO2 Incubator MCO-180IC Operation SOP

Figure 1: Incubator Components

1. Outer Door: sticks to frame with magnetic seal; door heater installed in the door panel
2. Tray Support: can be removed by lifting the front side and pulling it toward the door
3. Top Duct: inlet area for circulating air
4. Door Switch: detects door position (open/closed); stops the fan and CO2 gas feed
5. Circulation Fan: located inside the rear duct; fan can be autoclaved
6. Rear Duct: flow path for circulating air output; removable for access to circulation
7. Side Support: right and left supports can be removed for cleaning or disinfecting
8. Inner Door: made of tempered glass, avoid excessive impact on the glass
9. Tray: structure for support of samples and stir plates.
10. Water Level Sensor: detects the water level in the humidifying pan; located under the
humidifying pan cover
11. Humidifying Pan Cover: prevents spills from contaminating the humidifying pan
12. Humidifying Pan: properly installed when resting on the floor of the incubator cabinet and
covered by the humidifying pan cover; only use biopure water to fill the pan
13. Power Switch: this is the main power switch for the incubator
14. Air Sampler Outlet Cap: always attached except at time of air sampling
15. Air Sampler Outlet: use this outlet for air sampling; this also functions as an internal gas
outlet; should be covered with cap
Great Bay Community College Document Number: 1.5.3
320 Corporate Drive Revision Number: 3
Portsmouth, NH 03801 Effective Date: 18Jul08
Page 4 of 4
Title: Sanyo CO2 Incubator MCO-180IC Operation SOP

Figure 2: Control Panel

1. UV Indicator (this model is not equipped with UV)

2. Heat Indicator: this light comes on when the heater is energized
3. Digital Temperature Indicator: indicates current chamber operating temperature
4. CO2 Inject Indicator: this light comes on when CO2 gas is being injected
5. Digital CO2 Density Indicator: indicates current percentage of CO2 in the chamber
6. Calibration Key: not necessary to touch
7. Alarm Buzzer: press the “buzzer” key to silence the alarm when needed
8. CO2 Gas Supply Line Indicator (not installed in this model).
9. CO2 Gas Supply Line Switching Key (not installed in this model).
10. Enter Key: pressing this key inputs the set value into the controller
11. Numerical Value Shift Key ▲ ▲ : press this key in the set mode to change the numerical value
12. Digit Shift Key ►►: Press this key in the set mode to shift the position of the digit
13. Set Key: Press this key to access to the set mode
14. Over-heat Indicator: this light comes on when the chamber temperature reaches the upper
limit set value
15. Upper Limit Regulator: This regulator is used to set the upper temperature limit
16. RH Pan Indicator: this flashes when water in the humidifying pan is less than ~1 liter
17. Door Indicator: this light comes on when the door is open