| Popis: |
The oxygen gradient in the intestine influences intestinal physiology, and the microbial environment. Neurons that innervate the intestine are in constant communication with the local microbiome and this communication is regulated by the physiological health of the tissue. Measuring the communication between neurons and resident gastrointestinal cells could provide essential information about gut health. Despite this, a method to simultaneously culture intestinal tissue slices with a physiologically-relevant oxygen gradient with the capability of monitoring neural-gut communication in real-time has not been explored. Here, we designed and fabricated a 3D printed microfluidic device with precisely placed 500 μm delivery ports to maintain the oxygen gradient. The gradient is maintained from outlets below while allowing access to the slice from above for neurochemical detection with fast scan cyclic voltammetry and carbon-fiber microelectrodes. Outlets are placed in an oval where deoxygenated media is delivered to the middle of the slice, then oxygenated media is delivered to the outside of the slice. An oxygen sensitive fluorescent dye, Tris(2,2’-bipyridyl)dichlororuthenium(II), is used to characterize the tunability of the gradient delivery on a slice. Then while maintaining the oxygen gradient tissue function and viability were examined with fast scan cyclic voltammetry paired with carbon fiber microelectrodes by measuring transient release of neurotransmitters in the intestine. This chip allows direct access to intestinal slices for real time measurements, and imaging while maintaining an oxygen gradient to recreate the physiological environment which has not previously been done giving new insights to neuro-immune communication. |
