Islet Core

Perifusion

Advanced Diabetes Research

The Islet Core provides consultative advice and state-of-the-art equipment and services for the isolation and study of pancreatic islet structure and function in support of diabetes-related research.

Access Our Services in MiCORES

How We Serve Your Research Needs
  • Pancreas
  • Embryonic rudiments (E10.5 — birth)
  • Islet isolation

Embryonic tissue dissection

embryonic-dissections
At chosen embryonic stages, the pancreas can be isolated from developing mice or rats. Early stages can also be put in culture embedded in a collagen.

Tissue harvesting and islet isolation

Islet-isolation
We offer pancreas dissections and islet isolation from both rats and mice.
  • β- α-cell mass
  • Proliferation
  • Apoptosis

β-cell mass

5 sections spread throughout the pancreas;

  • Fluorescent Immunohistochemistry staining for insulin
  • Imaging
  • Morphometry
Beta Cell mass calculation

Immunostaining

In addition to insulin and glucagon, a number of antibodies for pancreatic markers are available through the Core.

islets-staining
Immunostaining for insulin (blue), glucagon(red) and amylase (green) on a paraffin embedded pancreatic section.
beta-cell-mass-multiple-images
Immunostaining for insulin (red) for five randomly selected sections at different depths within the pancreas.

Software assisted nuclei counting

  • Proliferation (Ki67)
  • Apoptosis (TUNEL) staining

In vivo imaging of fluorescent reporters in anesthetized mice

  • Imaging during the course of a glucose tolerance test
  • Image processing for analysis
  • Islet tracking over the course of the glucose challenge
fluorescent-reporter-live-tracing
A. Schematic representation of the sfGFP-CPep reporter mouse model. B. The pancreas is externalized through laparotomy on an anesthetized mouse, and placed a heated slide for imaging throughout an IPGTT. C. Glucose measurements during the course of the glucose challenge. D. Fluorescence pancreas imaging from a CPep-sfGFP mouse (Top: fluorescence, Bottom: Intensities are presented in pseudo-colors from dark blue to yellow). E. Quantification of individual response of the islets over the course of the glucose challenge (Left: normal scale, and Right: Log scale showing fold changes independently of islet size).
fluorescent-reporter-quanditifcation

AI-based image analysis

Using Aivia (Microscopy and Imaging Core), we can automate morphometric assessments (β- α-cell mass, proliferation and apoptosis) as well as fluorescent islet tracking and signal quantification after neural network training.

AI-based-morphometry

• Static GSIS
• Perifusion
• Respirometry

Static Glucose-stimulated insulin secretion

In culture with transwell inserts

Low glucose, high glucose, low glucose, KCI
Two pictures of diabetic lab equipment
Culture plate (left) with transwell inserts (close-up on the right)

 

Perifusion

Up to 12 chambers can be used in parallel to measure islet secretions using the custom program of your choice.

The perifusion chambers and reagents are temperature-controlled. The reagents can be gazed with CO2.

A series of images with lab equipment and cell data
Top left: Immunostaining for β- (blue) and α-cells(red). Top right: Secretion profile for islets after a perifusion assay. Bottom left: Dissecting microscope. Bottom right: Perifusion apparatus.

Respirometry (BaroFuse)

Respirometry and perifusion can run concomitantly on up to 6 samples at the same time.

A picture of the barofuse instrument
The BaroFuse instrument
A graph regarding mitochondrial respiration and insulin secretion in mouse islets

Simultaneous parallel measurements of mitochondrial respiration and insulin secretion in mouse islets. Representative measurement of oxygen consumption (top) and insulin release (bottom) from islets studied on the BaroFuse multichannel respirometry/perifusion system following infusion of glucose, nimodipine, and potassium cyanide. Top graph is the mean oxygen consumption (±SD) of two independent channels (75 islets each from islets isolated from the same mouse). Bottom graph is insulin secretion collected from the same islets from each independent well (blue, red).

Transfection

Islets can be transfected in vitro, or in vivo after I.P. delivery

islets-transfections
A. Transfection of islets in vitro, and B. 2 weeks after I.P. delivery of 1.10^12 particles of β-cell specific AAV8-mIP (mouse insulin promoter)-eGFP vector.

Islet dissociation and pseudo-islets

  • Islet dissociation
  • MACS separation of the β-cells
  • Transfection of single cells
  • Pseudoislet generation

 

pseudoislets
Generation of human pseudoislets following high efficiency viral transduction. A. Brightfield (top), live cell confocal images (middle) and confocal images (bottom) of intact human islet and human pseudoislet transduced for 2 hrs with adenoviral particles expressing eGFP under control of the PDX1 promoter for #-cell specific expression. B. Insulin secretion measured after static incubation in low (LG; 2 mM) and high glucose (HG; 20 mM), from transduced intact human islets or human pseudoislets from the same donors. C. Immunostaining for insulin, glucagon, and somatostatin of fixed and cryoembedded human pseudoislet sections.
  • Islet isolation
  • Secretion assays
  • Morphometry
  • Experimental design
  • sfGFP-CPep imaging
  • Embryonic rudiments cultures
  • Grafts and Imaging in the anterior chamber of the eye

Key Personnel

Scott Soleimanpour, MD

Scott A Soleimanpour, MD
Larry D Soderquist Professor
Professor of Internal Medicine
Professor of Molecular and Integrative Physiology and Associate Director
M-Diabetes for Type I Diabetes Basic Research
Metabolism
Endocrinology and Diabetes
Medical School
Corentin Cras Meneur

Corentin Cras-Meneur
Associate Research Scientist
Internal Medicine
Medical School
Emily Walker photo

Emily M Walker
Center Member

Contact Us
If you are an external customer, contact the core for pricing.