Hypoxia Core

Director

Imad Al Ghouleh, PhD | ima6@pitt.edu

The animal hypoxic core is located in the SBST 10th floor animal facility. The core contains 8 mouse chambers and 2 rat chambers. Each mouse chamber can hold 4 standard mouse cages with 4 mice per cage. Each rat chamber can hold 4 standard rat cages with 2 rats per cage.

Our core features ProOx 110 oxygen controllers and A-Chamber animal chambers from BioSpherix Ltd. This system allows precise setpoint and maintenance of hypoxic conditions for both acute and chronic animal studies. More information available at: https://biospherix.com/vascular-research

To minimize animal transfer between facilities, please be sure that SBST 10 location is in your IACUC protocol – this will allow you to order animals directly to SBST 10.

Please contact Dr. Imad Al Ghouleh (ima6@pitt.edu) if you have any questions.

1. For first time use please contact Dr. Imad Al Ghouleh (ima6@pitt.edu) to discuss your animal model needs and plan your experiments. You will be given instructions how to access the calendar to reserve chambers. You will also be given instructions how to use the chambers.

2. Prior to each experiment, please log into Google Calendar using the egdglab account in order to view the calendars and make reservations/changes. Once logged in, you may view the individual chamber calendars by clicking through the items on the left sidebar. When you make a new reservation, your calendar entry must follow this format: “PI-Last-Name/User’s Name/number of cages” (without quotes – for example, Gladwin/Courtney/4).

3. Once you made your reservation, please contact Dr. Imad Al Ghouleh (ima6@pitt.edu) with a cc to Dr. Andres Pulgarin (JROCHA@pitt.edu) to inform us of your new entry request. Also, Please contact us at least 1 week before starting your experiment so that we can pre-calibrate the system for you.

4. Once calibrated, you will be notified and you can place your animals in the chambers. Please note that opening the chamber will cause oxygen to normalize to atmospheric levels. Please make sure that you close the chamber door properly once you finish animal placement (and cage changes). Please also monitor the oxygen level to ensure it is decreasing back to the set point. If the oxygen level does not go back to the set point within 30 minutes, please contact the core.

IMPORTANT NOTE: Please do not manipulate the oxygen controller module under any circumstances. Core members will help you with any troubleshooting.

NOTE: To prevent over-booking and accommodate all users, if there are no animals in a pre-booked hypoxic chamber 2 days past the scheduled start date this reservation will be canceled by core personnel.

5. Animal care during hypoxia: Unfortunately, DLAR staff can’t provide animal care for animals under hypoxic exposure. Users are responsible for the care of those animals themselves. This includes monitoring and replenishing food and water, regular changing of cages and bedding, monitoring animal health and well-being, and filling in the animal husbandry log regularly.

6. Please be sure that your emergency contact info is available to DLAR staff. Please make sure that this information is easy to locate (best way is to print it and stick printout to the cubicle room door – cubicle/chamber numbers should be included).

7. At end of hypoxia exposure: Please clean the chamber once your experiment is over and your animals are out of hypoxia. Core use privileges may be revoked if chambers are not cleaned. Make sure to bring the empty tank back to BST12 storage space (see below) and notify Hypoxia Core members that your experiment is over, chamber has been cleaned and empty tank returned.

8. Nitrogen tank rotation: The low level of oxygen in hypoxic chambers is maintained by automated addition of nitrogen gas. It is the user’s responsibility to ensure nitrogen tanks do not run out during their experiments. If tanks are allowed to become empty oxygen levels would normalize compromising the quality of your experiment and results.

Based on our previous experience, to maintain a constant 10% oxygen level, nitrogen tanks needs to be replaced every 48 hrs for each mouse chambers and every 24 hrs for each rat chamber.

Users (and PI lab members) are responsible for transferring nitrogen tanks between BST12 and SBST10, monitoring nitrogen levels in tanks attached to their chambers, and changing tanks attached to their chambers as needed. Users are also expected to place 2 new tanks per chamber prior to pre-calibration.

Nitrogen tanks supply is kept at BST12 (12^th floor tank room near the freight elevators). Space is limited and timely exchange of empty tanks is needed. When you exchange tanks for your chambers, please make sure to return empty tanks back to BST12 by 3 pm on the day you changed it. Please do not wait untill both tanks in vivarium are empty in order to ensure continuous nitrogen supply and prompt tank rotation for your experiment.

Upon returning an empty tank, please make sure the tank cap is secured back on and a yellow “empty” card tag is placed on the tank. Make sure the tank is tightly secured in place. Please also make sure to log your tank usage and PI/user names on the log sheet next to the tanks in BST12.

Please contact Dr. Charles McTiernan (mctiernanc@upmc.edu) with any nitrogen questions.