BTX AgilePulse ID User Manual page 26

AgilePulse ™
Injection
• Prepare transfectant and adjuvant if desired in 0.25 to 1X PBS.
Suggested plasmid DNA concentration range is 1 to
4 μg/ml (recommended starting concentration is 2 μg/ml).
• To aid in accurate administration of the procedure and
minimize the potential for startle reflex, anesthetize animal
as per veterinarian recommendations. The subject should be
anesthetized to a level where it is not responsive to a strong
toe pinch or other equivalent stimulus.
• Clip hair or depilate to expose target tissue.
• Clean site with 70% alcohol and remove excess alcohol
with gauze.
• Anesthetized animal should be oriented to allow for easy
exposure of target tissue.
• Obtain tuberculin syringe(s), 3/10 cc (BD model 328431),
loaded with transfectant mixture. Use a 30 gauge needle For
muscle electroporation, fit the syringe with P10 tubing to
ensure an even depth of insertion into the muscle targeted.
• For skin electroporation insert needle tip 2-3 mm just under
the skin (needle should be visible) and slowly inject DNA into
site. A clear bleb in the skin should be visible post-injection.
• For muscle electroporation insert needle tip into the muscle at
desired depth and slowly inject DNA into site.
• Inject 20 to 30 μl transfectant solution.
• Repeat the procedure on the other sites if necessary.
24
ID Tutorial
AgilePulse ID In Vivo Gene Delivery System
Publication 015-101444 Rev 2.0 • www.btxonline.com
Electroporation
• Insert needle electrode so the injection site is placed
between the two needle rows. Begin the process by pressing
the "Next" button. This will take you to a screen with a series
of buttons.
• Press the "Load" button. Read the "load reading" on the right
hand side of the screen. If it is below 100 ohms, then the
electrode is inserted far enough. If above 2000 ohms, insert
the needles deeper in the tissue.
• After correct insertion has been verified, sequentially press
the remaining buttons as they fill-in to start electroporation
sequence.
• Keep pushing electrode firmly against animal until pulsing
stops.
• Remove electrode after delivery of pulses.
• Check the log file to verify proper delivery.
• Repeat electroporation as needed, then press "Done" icon
to return to the main screen.
• Observe animal and keep warm until it recovers from
anesthesia. Put animal back into its cage.