Intestinal Adaptation Laboratory

Laser Capture Microdissection (LCM) Microscopy 

A significant and novel aspect of our laboratory is the ability to employ laser capture microdissection (LCM) to successfully extract enterocytes from frozen tissue sections of mouse ileum and to measure differences in gene expression along the crypt/villus axis within these captured cells. In our laboratory, we use an ArcturusXT Veritas microdissection instrument to capture cells from the mouse small intestine, as shown, by the crypt enterocytes isolated from the mouse ileum. The subepithelial structures section has remained intact along with the adjacent enterocytes that were not taken. 

The RNA from these captured cells is then extracted, and specific genes are used to confirm each cell compartment. Using real-time polymerase chain reaction (PCR), we have determined the mRNA expression level in LCM-captured material for a number of genes. In comparing mice that have undergone sham operation versus small bowel resection (SBR), we identified differences in postresection mRNA expression occurring within the proliferative compartment of the crypt enterocytes as well as villus enterocyte compartment. With linear amplification LCM, our lab also is using RNA to probe microarrays. Christopher Erwin, Ph.D., is director of the LCM core facility.

Related Publications
Knott AM, Erwin CR, Profitt SA, Juno RJ, Warner BW. Localization of postresection EGF receptor expression using laser capture microdissection. J Pediatr Surg 38: 440 - 445, 2003.

Stehr W, Mercer TI, Bernal NP, Erwin CR, Warner BW. Opposing Roles for p21waf1/cip1 and p27kip1 in Enterocyte Differentiation, Proliferation, and Migration. Surgery, 138(2):187-94, 2005.

Vikram Prasad, Gregory P. Boivin, Marian L. Miller, Lynne H. Liu, Christopher R. Erwin, Brad W. Warner and Gary E. Shull. Haploinsufficiency of Atp2a2, Encoding the SERCA2 Ca2+ Pump, Predisposes Mice to Squamous Cell Tumors Via a Novel Mode of Cancer Susceptibility. Cancer Research, 65(19): 8655-61.

Contact Us:
Brad W. Warner, M.D.
Pediatric Surgeon-in-Chief
St. Louis Children's Hospital
Apolline Blair St. Louis Children's Hospital Professor of Surgery
Chief, Division of Pediatric Surgery
Washington University School of Medicine

St. Louis Children's Hospital
Suite 5560 - Campus Box 8235
One Children's Place
St. Louis, Mo 63110
(314) 454-6022
E-mail brad.warner@wustl.edu 

Christopher R. Erwin, Ph.D.
Associate Professor of Surgery
Division of Pediatric Surgery
Washington University School of Medicine

St. Louis Children's Hospital
Suite 5560 - Campus Box 8235
One Children's Place
St. Louis, Mo 63110
(314) 454-6022
E-mail erwinc@WUSTL.EDU

Jun Guo, Ph.D.
Assistant Professor of Surgery
Division of Pediatric Surgery
Washington University School of Medicine

St. Louis Children's Hospital
Suite 5560 - Campus Box 8235
One Children's Place
St. Louis, Mo 63110
(314) 454-6022
E-mail guoj@wudosis.wustl.edu