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    Articles

    Nanofil

    Sub-microliter injection system for small animal research

    NanoFil is a specially designed 10 microliter syringe developed in response to customer requests for improved microinjection in mice and other small animals. It makes quantitative nanoliter injection much easier and more accurate than any other method currently in use. In traditional nanoliter injection, the dead volume of the syringe and needle is backfilled with oil as a displacement transfer. This makes the process messy and risks contamination of the injected sample. NanoFil’s low volume, flexible tubing eliminates the need for oil. Injection is now simpler, and less messy, and there is no possibility of oil contamination in critical applications such as ophthalmology research (please see the Retinal Pigment Epithelial (RPE) and Intra Ocular (IO) injection kits listed below. When the inner diameter of the tip on a conventional syringe is reduced to less than 100 micron, it is very difficult to back fill the solution at a reasonable speed. NanoFil solves this problem by using a tip coupling mechanism that makes it possible to change the syringe tip during the experiment. Simply load the sample using a larger tip, such as the 26 gauge needle provided with the syringe, and then replace it with a micro tip for sample injection. On a conventional 10 microliter syringe, a solid ring or bushing is permanently bonded to the tubing. Replacing the tip in middle of the experiment is not practical. With the nanoFil, different gauge tips can be exchanged on the syringe by a simple twist of the brass lock, gently pulling out the tip, and replacing with the desired new tip. To secure the tip, NanoFil uses an olive shaped silicon gasket that is similar to, but much sturdier than, some of the microelectrode holders used for electro physiology recording. The silicone gasket makes it possible to hold not only metal tips but also glass and quartz tubing. Many types of tubing can be easily connected to the syringe as long as the outer diameter (OD) is close to, but not more than, the inner diameter (ID) of the inside barrel. Specially designed tips The application potential of this design is tremendous.

    Flexible quartz capillaries used in Gas Chromotography (GC) and Capillary Electrophoresis (CE) can also be easily coupled to the syringe. Specially designed tips as small as 36 gauge (110 micron OD) are offered in both blunt and bevelled styles. Our studies have shown that these tips will cause less trauma to the tissue than any other form of micro syringe currently in use. NanoFil has a unique coupling mechanism that allows many different forms of small tubing and tips to be coupled with the syringe barrel.

    Based on application requirements, NanoFil can be used in several different configurations Direct injection by hand: This is the simplest and most economical way to inject. Any of our tips can be inserted directly into the NanoFil syringe. Even the SilFlex tubing can be inserted to switch from hand injection to the other methods listed below. The limitation of this method is the difficulty achieving sub microliter resolution. Installed on WPI’s UMP-II microsyringe pump: This will allow the user to achieve nanoliter resolution and reproducibility. For neural system injection, mount the UMPII on a stereotaxic frame. SilFlex tubing and holder: The needle is mounted on a small plastic holder that is connected to the NanoFil by a 35 cm length of flexible tubing. The NanoFil is mounted on the UMP II pump. This configuration allows the user to hold the animal in one hand and insert the needle with the other. When the needle reaches the desired location, activate the pump using the footswitch and the pre-programmed injection volume will be delivered.This configuration gives a nanoliter level of accuracy and reproducibility. It is best suited for applications such as the RPE and IO injection.

    Selecting the correct tip for your application

    The replaceable needles used with the NanoFil are available with either blunt or bevelled tips. The blunt tip is used for injection into soft tissue and when a uniform solution distribution is needed. The bevelled style is used for applications that involve the penetration of a tough tissue. One of the main factors that limit the resolution and accuracy of conventional micro syringes to the upper tens of nanoliters range is diffusion in the large tip ID. When the tip ID is equal or larger than 100 micron, the error caused by tip diffusion is in the nanoliter range level ( (100 micron)^3 = 1 nanoliter ). With a 36 gauge needle installed on the NanoFil, the error caused by diffusion will be reduced to the sub nanoliter level, making accurate injection of a nanoliter possible. All of WPI’s bevelled tips have a unique 25 degree tri-surface bevel that is optimized for microinjection. A 10 degree single-surface bevelled tip penetrates better than one with a 25 degree angle, however the distance between the upper opening to the tip (see section “F” in Fig 2) is longer. As a result, it requires a deeper penetration of the tip to achieve the same level of liquid delivery. Deeper penetration means more tissue damage. WPI’s unique 25 degree bevelled tip solves this problem with two extra bevelled surfaces. The tip of a single surface bevelled tip is actually a blade instead of a point. It dulls very quickly. In contrast, the trisurfaced tip has a real point. It not only penetrates much better but is also much more durable. Our tests show that our 33 gauge, 25 degree bevelled tip penetrates easier and lasts longer than other manufacturer’s 33 gauge, 10 degree single bevelled tips. With a 35 gauge tri-surface bevelled tip, the resistance to the penetration becomes even less. Each of our tips undergo a penetration test before leaving the factory to guarantee the best results for our customers.