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Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes.

Publication ,  Journal Article
Couter, CJ; Surana, NK
Published in: J Vis Exp
May 8, 2016

The intestines - which contain the largest number of immune cells of any organ in the body - are constantly exposed to foreign antigens, both microbial and dietary. Given an increasing understanding that these luminal antigens help shape the immune response and that education of immune cells within the intestine is critical for a number of systemic diseases, there has been increased interest in characterizing the intestinal immune system. However, many published protocols are arduous and time-consuming. We present here a simplified protocol for the isolation of lymphocytes from the small-intestinal lamina propria, intraepithelial layer, and Peyer's patches that is rapid, reproducible, and does not require laborious Percoll gradients. Although the protocol focuses on the small intestine, it is also suitable for analysis of the colon. Moreover, we highlight some aspects that may need additional optimization depending on the specific scientific question. This approach results in the isolation of large numbers of viable lymphocytes that can subsequently be used for flow cytometric analysis or alternate means of characterization.

Duke Scholars

Published In

J Vis Exp

DOI

EISSN

1940-087X

Publication Date

May 8, 2016

Issue

111

Location

United States

Related Subject Headings

  • Peyer's Patches
  • Mice
  • Lymphocytes
  • Intestine, Small
  • Intestinal Mucosa
  • Flow Cytometry
  • Animals
  • 1702 Cognitive Sciences
  • 1701 Psychology
  • 0601 Biochemistry and Cell Biology
 

Citation

APA
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ICMJE
MLA
NLM
Couter, C. J., & Surana, N. K. (2016). Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes. J Vis Exp, (111). https://doi.org/10.3791/54114
Couter, Cheryn J., and Neeraj K. Surana. “Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes.J Vis Exp, no. 111 (May 8, 2016). https://doi.org/10.3791/54114.
Couter, Cheryn J., and Neeraj K. Surana. “Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes.J Vis Exp, no. 111, May 2016. Pubmed, doi:10.3791/54114.

Published In

J Vis Exp

DOI

EISSN

1940-087X

Publication Date

May 8, 2016

Issue

111

Location

United States

Related Subject Headings

  • Peyer's Patches
  • Mice
  • Lymphocytes
  • Intestine, Small
  • Intestinal Mucosa
  • Flow Cytometry
  • Animals
  • 1702 Cognitive Sciences
  • 1701 Psychology
  • 0601 Biochemistry and Cell Biology