Abstract
A microfluidic islet perifusion device was developed for the assessment of dynamic insulin secretion of multiple islets and simultaneous fluorescence imaging of calcium influx and mitochondrial potential changes. The device consists of three layers: first layer contains an array of microscale wells (500 μm diameter and 150 μm depth) that help to immobilize the islets while exposed to flow and maximize the exposed surface area of the islets; the second layer contains a circular perifusion chamber (3 mm deep, 7 mm diameter); and the third layer contains an inlet-mixing channel that fans out before injection into the perifusion chamber (2 mm in width, 19 mm in length, and 500 μm in height) for optimizing the mixing efficiency prior to entering the perifusion chamber. The creation of various glucose gradients including a linear, bell shape, and square shapes also can be created in the microfluidic perifusion network and is demonstrated.
Highlights
3. 25-30 mice islets were incubated with 5 μM Fura-2/AM and 2.5 μM Rhodamine 123 (Rh123, a mitochondrial potentials indicator, Sigma, MO) for 30 min at 37 °C in Krebs-Ringer buffer (KRB) containing 2 mM glucose
After setup, the mice islets are perifused with KRB containing 2 mM glucose for 10 min and a glucose ramp (2mM 25mM) for 25 min
Changes in Mitochondrial potentials are seen earlier as expected, at about 11 minutes. This data demonstrates the advantage of using this microfluidic network to characterize islet physiology
Summary
[NOTE: do not hold the wafer with tweezers after spinning SU8] 3. Soft bake the wafer at 65 °C for 20 min and at 95 °C for 50 min. Microchannel Master Protocol (500 μm deep wells) 3. Soft bake the wafer at 65 °C for 15 min and at 95 °C for 2 hrs and 30 min.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
