Frequently Asked Questions About the Neuron Device at Xona microfluidics, LLC

How do I place an order?

To place an order, please download our order form (you will need the Adobe Acrobat Reader) and then simply fax it to us at:

(951)308-2488 or send it via email to xona@xonamicrofluidics.com

What material are the Neuron Devices made from?

A. Polydimethylsiloxane (PDMS)

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Is it safe?

A. Yes, PDMS is considered biologically inert and safe for both humans and cells.

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Does PDMS interfere with microscopy and live cell imaging?

A. PDMS is optically transparent and suitable for most types of microscopy and cell imaging.

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What is the purpose of the neuron microfluidic device?

A. The neuron microfluidic device allows researchers to compartmentalize neuronal cultures. The neuron device consists of two culture chambers separated by a microgroove barrier. Neurons are typically loaded on one side of the device. The size of the microgroove barrier allows axons to grow through the barrier while keep the cell bodies (Soma) isolated from the distal chamber. Please note however that the microgrooves will not keep migrating cells from crossing the barrier. In addition high density cultures may result in some cell bodies entering the microgrooves.

The neuron device also makes it possible to perform axotomy, cutting axons via vacuum aspiration in the distal chamber and observing their regrowth.

The neuron device is also small in size (total volume ~ 600 ul) thus researchers save on the expensive reagents needed for neuronal culture.

What is fluidic isolation?

A. The neuron device consists of two chambers 100 um in height connected by a microgroove barrier that is 3 um in height. The hydrostatic pressure between the two chambers separated by the microgroove allows the researcher to fluidically isolate each chamber. This is accomplished by keeping the volumes in the wells on one side of the device higher then the other side of the device. The difference in volume creates hydrostatic pressure thus fluidically isolating each compartment.

Can immunohistochemistry be performed in the device?

A. Yes, samples can be stained and imaged in the device. In addition, if non-plasma bonding was performed, it may be possible (though not always necessary) to remove the device for imaging. In the case of non-plasma bonding, chilling the device for 5 minutes on ice after fixing helps with removing the device while minimizing damage to the cells.

E18 Cortical Rat Neuron stained with Map2

E18 Cortical Rat Neuron stained with Map2

Can protein lysate be isolated from the device?

A. Yes, however keep in mind that in most cases 80,000 to 200,000 cells are being loaded per device. Thus the amount of protein lysate obtained may or may not be suitable for your application. The amount of protein lysate from the axonal compartment will be even lower. If you seek to do a western blot with an antibody to a protein known to be low in abundance with lysate from the axonal compartment one device may not yield enough protein. It is possible to pool lysates from several devices however Xona Microfluidics LLC makes no guarantees that you will be able to detect your protein of interest.

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Can RNA be isolated from the device?

A. Yes, however as with protein, the low amount of cells in the device generally translates into low yields of RNA. Like for protein lysates, devices can be pooled in order to obtain enough RNA to detect low yield RNA’s. Your success in isolating RNA will depend on the abundance of your target RNA.

Product Request/Packing of our Devices

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What is plasma bonding?

A. Plasma bonding involves using a plasma cleaner to produce reactive oxygen species on the surface of both the PDMS and glass. In addition to cleaning the glass and PDMS the plasma cleaner also sterilizes. After treating with plasma when the PDMS comes in contact with the glass it forms a tight irreversible bond. Plasma treating also makes the surfaces hydrophilic thereby facilitating the introduction of liquids into the device. We typically recommend the Harrick Plasma cleaner. Please go here for more information on plasma cleaning.

Do I need a plasma cleaner?

A. No, the neuron device can be bound to cover glass without using a plasma cleaner. It is however important to use clean, sterile glass. Typically we recommend using a water bath sonicator to clean the glass followed by an ethanol bath to sterilize them. After removing from ethanol we allow the cover glass to dry in a biosafety cabinet in order to maintain sterility.

Also, the neuron devices will need to be sterilized prior to use. This can be done by rinsing the devices with 70% Ethanol and allowing them to dry in a biosafety cabinet. The devices can also be sterilized by autoclaving. However, one needs to take care in choosing what type of container to autoclave the devices in. Some plastic containers and bags can impart toxins into the device which will then kill the cells. Please go here for more information on plasma cleaning.