More recently, through collaboration with Dr. Fang Huang at Purdue University, the lab has begun to address how changes in the structure and composition of key β cell organelles contribute to the loss of insulin secretory capacity in the β cell during diabetes development. Dr. Huang's group has developed a novel super resolution imaging technique known as whole-cell 4Pi single-molecule switching nanoscopy (W-4PiSMSN) which allows for 10- to 20-nm 3D resolution across the thickness of the entire mammalian cell due to its ability to overcome the diffraction limit of light. The lab is using this technique to image the ultrastructural architecture of the ER in the β cell. By leveraging the nanometer resolution and localization capabilities of W-4PiSMSN, the precise meshwork of the ER can be visualized and the tubule diameter can be quantified.
[Pictured: Representative super-resolution image of the rat insulinoma-derived pancreatic β cell line with the endoplasmic reticulum labeled using a Sec61β-GFP plasmid and GFP-Booster Alexa Fluor 647 nanobody (shown in orange). Scale bar = 5 micrometers. Photograph courtesy of Madeline McLaughlin, Fang Huang, PhD, and Carmella Evans-Molina, MD, PhD.]
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