ACT-LABS gun system Instruction Manual download pdf (Page 35)

5. Incubate 30 min to 4 hr under the appropriate conditions.

6. Prepare the Helios Gene Gun for operation as described in Section 5.3.

7. Immediately prior to DNA delivery, aspirate the media from the dish.

8. Hold the dish perpendicular to the spacer and touch the end of the plastic spacer (if the spac-

er is sterile), or as close as possible to the target, and discharge the Gene Gun (Figure 18).

9. Add 1.5–2 ml of media to the plate and return to the incubator.

6.3

In vivo

Delivery to Epidermis

Animal Preparation

1. Anesthetize the animal if necessary for safe handling.

2. Clip fur as closely as possible over the desired target area using Oster clippers with a

#40 surgical blade and brush or vacuum fur off.

3. (Optional) After clipping, a commercial depilatory such as Nair can be used to completely

remove the animal’s fur. This treatment removes the stratum corneum from the skin, com-

pletely exposing the epidermis. Carefully rinse the skin with warm water following

depilatory treatment.

4. If the target site is wet or dirty, clean and dry with 70% ethanol.

Helios Gene Gun Operation

1. Prepare the Helios Gene Gun for operation as described in Section 5.3.

2. Hold the spacer directly against the target site (Figure 19) and discharge the device.

(Discomfort following Helios Gene Gun bombardment of the skin is minimal. No appar-

ent macroscopic disruption of the skin, external bleeding or hematoma should be observed.

Some animals may show transient erythema or inflammation at the treatment site.)

Fig. 19.

In vivo

bombardment of murine epidermis (photo courtesy of Auragen, Inc.).

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ACT-LABS gun system Instruction Manual Page 35