Bitan:make subQ pumps

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Making Subcutaneous pumps at the VA

I) Do we have enough of all materials?

II) To make 0.9% saline A) 100ml dd H2O, 0.9g NaCl

III) to sterilize/filter saline for pump incubation bath

A) filters with blue lids B) spray gloves with 70% EtOH in hood, open sterile filter bag in hood, C) attach vacuum tube to filter and turn vacuum/yellow handle on. D) Pour in the 0.9% saline slowly E) Once saline all through, turn vacuum off and put filter lid on F) Label lid with saline and group name of animals

IV) To make pumps in the hood

A) Get 1cc or .5cc syringes with no needle B) Open miniosmotic pump parts in hood: tube and cap and blunt needle C) Put blunt needle on syringe and pull up 400ul of drug/vehicle solution for 3 pumps (each pump model 1004 holds 100ul) D) make sure top of pump is dry (with kimwipe) stick needle into pump slowly and hold at an angle so hole of pump is not totally blocked by needle E) look at starting amount of liquid in syringe, push syringe pump in slowly until a liquid bubble forms at the hole of the pump, make sure about 100ul have been used 1) if 100ul haven’t been used, you could dry top of pump and keep filling, should see air bubbles come up until pump is full. 2) Might have to get new pump if can’t figure out F) Take cap with metal catheter and place into hole. Push sides of cap down with 2 fingers without covering hole in cap. If 100ul in pump, small amount of liquid should come out of cap. G) Drop into sterile saline, should sink if pump is filled accurately. H) Put in 37 deg incubator at least over night to prime. 1) Could prime over weekend if need to (Sally Frautschy) 2) Franziska Richter puts 1ml of sterile saline into 1.5ml tubes and incubates each pump separately.

(Tweezer is in hallway freezer in the door)