Quantitative measurement of zinc secretion from pancreatic islets with high temporal resolution using droplet-based microfluidics.
We assayed glucose-stimulated insulin secretion (GSIS) from live, murine islets of Langerhans in microfluidic devices by the downstream formation of aqueous droplets. Zinc ions, which are cosecreted with insulin from beta-cells, were quantitatively measured from single islets with high temporal resolution using a fluorescent indicator, FluoZin-3. Real-time storage of secretions into droplets (volume of 0.470 +/- 0.009 nL) effectively preserves the temporal chemical information, allowing reconstruction of the secretory time record. The use of passive flow control within the device removes the need for syringe pumps, requiring only a single hand-held syringe. Under stimulatory glucose levels (11 mM), bursts of zinc as high as approximately 800 fg islet(-1) min(-1) were measured. Treatment with diazoxide effectively blocked zinc secretion, as expected. High temporal resolution reveals two major classes of oscillations in secreted zinc, with predominate periods at approximately 20-40 s and approximately 5-10 min. The more rapid oscillation periods match closely with those of intraislet calcium oscillations, while the slower oscillations are consistent with insulin pulses typically measured in bulk islet experiments or in the bloodstream. This droplet sampling technique should be widely applicable to time-resolved cellular secretion measurements, either in real-time or for postprocessing.
Easley
Christopher J
CJ
Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, Vanderbilt University, 702 Light Hall, Nashville, Tennessee 37232, USA.
Rocheleau
Jonathan V
JV
Head
W Steven
WS
Piston
David W
DW
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D044085:Microfluidics / Q000295:instrumentation* / Q000379:methods; D011083:Polycyclic Compounds / Q000737:chemistry*; D015032:Zinc / Q000378:metabolism*
C451182:FluoZin-3; D007328:Insulin; D011083:Polycyclic Compounds; D015032:Zinc; D002118:Calcium