Gnotobiotic Mouse Technology: An Illustrated Guide

Chriss J. Vowles, Natalie E. Anderson, Kathryn A. Eaton

October 28, 2015 by CRC Press
Reference - 237 Pages - 275 Color Illustrations
ISBN 9781498736329

Features

Summary

The popularity of germ-free animal models, particularly mice, for investigation of human physiology and disease has recently exploded. Gnotobiotic Mouse Technology: An Illustrated Guide provides the first manual for the maintenance, husbandry, and experimental manipulation of germ-free and gnotobiotic mice. It includes information on all aspects of establishing and operating a germ-free mouse research facility, from basic principles and equipment to detailed instructions for assembling and maintaining isolators, sterilizing supplies, handling animals, and monitoring sterility.

The book breaks down techniques and procedures into modules, each of which contains a step-by-step practical and visual guide to a set of related procedures. Each description consists of a materials list, introduction, and general overview, followed by a thoroughly illustrated walkthrough of the steps for each procedure. The descriptions end with troubleshooting tips. Technical chapters are heavily illustrated and include notations of potential pitfalls and alternatives. Other chapters discuss management procedures and practices such as the hiring and training of personnel, setting fee schedules, and record keeping.

This book is a landmark resource for establishing and maintaining a facility for germ-free mouse research. Useful to both technicians and investigators, it presents every step necessary to establish a successful facility. It also gives direction in expanding multi-user facilities and applying new technologies to your current practices.

Table of Contents

A Brief History of Germ-Free Life
References

Overview of Gnotobiotic Technology
Overview
How Do We Get Germ-Free Mice?
Germ-Free Housing and Barrier Types
References

Equipment and Terminology

Personal Protective Equipment
Introduction
Types of PPE
Applying Sterile PPE

Sterilants and Sterilization
Introduction
Sterilization Methods
Cold Sterilization
Wrapping Materials for Autoclaving
Gas Sterilization (Ethylene Oxide)

Isolator Setup
Introduction
Part 1: Small (5.5 x 3 feet) Isolator Assembly
Part 2: Large Isolator Assembly

Port Entry and Exit
Introduction
Supplies
Preparing, Opening, and Closing the Port
Proper Placement of Materials

Sterilizing Food and Supplies
Introduction
Biological Indicators
Assembling Supply Cylinders: Installing HEPA Filtration Media
Supplies
Prepare the Cylinder
Fill the Cylinder
Water Sterilization

Supply Cylinder Entry
Introduction
Supplies
Procedure

Isolator Maintenance
Introduction
Part 1: Cleaning and Maintenance
Part 2: Replacing Gloves on an Active Isolator
Part 3: Repairing Holes in Isolators
Part 4: Working in the Isolator

Aseptic Mouse Transfer
Introduction
Part 1: Internal Transfer between Isolators
Part 2: External Transfer of Mice

Working with Germ-Free or Gnotobiotic Mice in the Class II Biosafety Cabinet
Introduction
Supplies
Experimental Manipulations in the Biosafety Cabinet

Shipping Mice
Introduction
Procedure

Rederivation
Introduction
Derivation by Cesarean Section
Derivation by Embryo Transfer
References

Microbiological Testing
Introduction
Serologic Screening
Part 1: Bacterial Culture
Part 2: Examination of Gram-Stained Slides for the Presence of Bacteria
References

Genetic Testing
References

Record Keeping
Introduction
Colony Management Documents
Project-Planning Documents
Other Documents

Facility Setup and Management
Introduction
Establishing a New Facility

Appendix: Sources for Equipment and Supplies

Author(s) Bio

Chriss J. Vowles

co-manages a multi-investigator germ-free research laboratory at the University of Michigan in Ann Arbor. He began his career at the University of Michigan in 2003, working full-time as a husbandry technician in the Unit for Laboratory Animal Medicine. In 2006, he discovered gnotobiotic technology. At that time, the Germ Free Laboratory was just starting. Chriss joined the research group on the ground floor and has been growing with it ever since. The idea of maintaining a germ-free, complex organism fascinated him then and still captivates him today. After eight years of maintaining a germ-free colony of mice, the laboratory is still evolving, he is still learning, and his trials, tribulations, and rewards are constant.

Natalie E. Anderson

is a research technician lead at the University of Michigan’s Germ Free Core. In 2009, she graduated from Michigan State University with a bachelor’s degree in animal science and accepted a position with SoBran’s contract for the National Institute of Allergy and Infectious Diseases (NIAID). In December 2010, she returned to Michigan to take a position with the University of Michigan’s Unit for Lab Animal Medicine. Shortly thereafter, she started working part-time in the Germ Free Core and quickly learned to love the daily challenges involved in maintaining a germ-free colony. Natalie joined the core full time in July 2013.

Kathryn A. Eaton