Methods & Standards

 

Protocols

Clamp Manual (download)

A full step-by-step description of the surgery, isotope clamp method and isotope calculations as performed by the Vanderbilt MMPC.

Surgical

Experimental

Note: Select protocols are shown. Inquire with Dr. Lantier regarding other tests and protocols.

Online analysis tools

CalR: A Free Web Application for Indirect Calorimetry Analysis
CalR generates time, bar and regression plots from data generated by CLAMS, Sable Systems and TSE while providing optimal statistical analysis (ANCOVA)

Published Methods and Standards

Design and interpretation of metabolic studies

Hyperinsulinemic-euglycemic clamps in conscious, unrestrained mice.
Ayala JE, Bracy DP, Malabanan C, James FD, Ansari T, Fueger PT, McGuinness OP, Wasserman DH.
J Vis Exp. 2011 Nov 16;(57).
Approach to assessing determinants of glucose homeostasis in the conscious mouse.
Hughey CC, Wasserman DH, Lee-Young RS, Lantier L.
Mamm Genome. 2014 Oct;25(9-10):522-38.
Considerations in the design of hyperinsulinemic-euglycemic clamps in the conscious mouse.
Ayala JE, Bracy DP, McGuinness OP, Wasserman DH.
Diabetes. 55(2): 390-7, 2006.
NIH Mouse Metabolic Phenotyping Centers: the power of centralized phenotyping.
Laughlin MR, Lloyd KC, Cline GW, Wasserman DH; Mouse Metabolic Phenotyping Centers Consortium.
Mamm Genome. 2012 Oct;23(9-10):623-31.
Standard operating procedures for describing and performing metabolic tests of glucose homeostasis in mice.
Ayala JE, Samuel VT, Morton GJ, Obici S, Croniger CM, Shulman GI, Wasserman DH, McGuinness OP; for the NIH Mouse Metabolic Phenotyping Center Consortium.
Dis Model Mech 3 (9-10):525-534, 2010.
Glucose metabolism in vivo in four commonly used inbred mouse strains.
Berglund ED, Li CY, Poffenberger G, Ayala JE, Fueger PT, Willis SE, Jewell MM, Powers AC, Wasserman DH.
Diabetes. 57(7):1790-9, 2008.
NIH Experiment in Centralized Mouse Phenotyping: The Vanderbilt Experience and Recommendations for Evaluating Glucose Homeostasis in the Mouse.
McGuinness OP, Ayala JE, Laughlin MR, Wasserman DH.
Am J Physiol Endocrinol Metab 297(4):E849-855, 2009.
Lost in translation.
Wasserman DH, Ayala JE, McGuinness OP.
Diabetes 58(9):1947-50, 2009.

Tracer methodology

Dose-response curves for in vivo insulin sensitivity in individual tissues in rats.
Kraegen EW, James DE, Jenkins AB, Chisholm DJ:
Am J Physiol 248 :E353 –E362,1985.
Isotope Tracers in Metabolic Research: Principles and Practice of Kinetic Analysis,
2nd Edition. John Wiley and Sons, Inc., New Jersey. 2004.
Wolfe RR, Chinkes DL.

Indirect Calorimetry

CalR: A Web-Based Analysis Tool for Indirect Calorimetry Experiments.
Mina AI, LeClair RA, LeClair KB, Cohen DE, Lantier L, Banks AS.
Cell Metab. 2018 Jul 4. pii: S1550-4131(18)30401-7.
Find CalR here: https://calrapp.org/
Mathematical model for the contribution of individual organs to non-zero y-intercepts in single and multi-compartment linear models of whole-body energy expenditure.
Kaiyala KJ.
PLoS One. 2014 Jul 28;9(7):e103301.
Toward a more complete (and less controversial) understanding of energy expenditure and its role in obesity pathogenesis.
Kaiyala KJ, Schwartz MW.
Diabetes. 2011 Jan;60(1):17-23.
Identification of body fat mass as a major determinant of metabolic rate in mice.
Kaiyala KJ, Morton GJ, Leroux BG, Ogimoto K, Wisse B, Schwartz MW.
Diabetes. 2010 Jul;59(7):1657-66.

Intravital microscopy

Insulin exits skeletal muscle capillaries by fluid-phase transport.
Williams IM, Valenzuela FA, Kahl SD, Ramkrishna D, Mezo AR, Young JD, Wells KS, Wasserman DH.
J Clin Invest. 2018 Feb 1;128(2):699-714
Automated quantification of microvascular perfusion.
McClatchey PM, Mignemi NA, Xu Z, Williams IM, Reusch JEB, McGuinness OP, Wasserman DH.
Microcirculation. 2018 Jun 16:e12482.