This volume provides a variety of standard protocols used to cryopreserve or freeze-dry different types of specimens. In addition, it provides chapters focused on the fundamental principles of cryopreservation, vitrification, and freeze-drying. Several state of the art microscopic, spectroscopic as well as calorimetric methods are highlighted that can be used to study cellular and macromolecular changes in response to freezing or drying. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, Cryopreservation and Freeze-Drying Protocols, Third Edition serves as a practical guideline for studies on freezing and drying processes as well as preservation strategies for biological specimens.

PART I: Fundamental Aspects And Freezing Technology                                             

1       Principles of Cryopreservation                                                                   

David E. Pegg

2          Principles of Cryopreservation By Vitrification                                                         Gregory M. Fahy and Brian Wowk

3          Modeling and Optimization of Cryopreservation                                                       James Benson

4          The Principles of Freeze-Drying                                                                                 Gerald D.J. Adams, Isobel Cook, and Kevin R. Ward

PART II: Methods To Study Freezing and Drying Processes                                     

5.         Use of In Situ Fourier Transform Infrared Spectroscopy to Study Freezing and Drying of Cells                                                                                                     Willem F. Wolkers and Harriëtte Oldenhof

6.         Calorimetric Analysis Of Cryopreservation and Freeze-Drying Formulations          

Wendell Q. Sun

7.         Measurement Of Intracellular Ice Formation Kinetics By High-Speed  Video Cryomicroscopy                                                                                    

Jens O. M. Karlsson

8.         Laser Scanning Microscopy in Cryobiology                                                  Frank Stracke, Asger Kreiner-Møller, and Heiko Zimmermann

9.         Low Temperature Electron Microscopy                                                                     Roland A. Fleck

PART III: Cryopreservation Protocols                                                                             

10        Cryopreservation of Semen From Domestic Livestock                                              Harald Sieme, Harriëtte Oldenhof

11.       Cryopreservation of Mammalian Oocytes                                                                  Victoria Keros, Barry J. Fuller

12.       Vitrification: A Simple And Successful Method for Cryostorage of Human Blastocysts                                                                                                    

Juergen Liebermann

13        Efficient Cryopreservation of Human Pluripotent Stem Cells By

Surface-Based Vitrification                                                                                        Julia C. Neubauer, Axel F. Beier, Nils Geijsen, and Heiko Zimmermann

14        Cryopreservation Of Greenshell(TM) Mussel (Perna Canaliculus) Sperm                      Serean L. Adams, John F. Smith, Jolene Taylor, Lindsay T. Mcgowan, and

H. Robin Tervit  

15        Membrane Modification Strategies for Cryopreservation                             

Phillip H. Purdy and James K. Graham

16.       Sperm Clean-Up and Density Centrifugation Procedures                            

Harald Sieme and Harriëtte Oldenhof

17.       Cryopreservation of Red Blood Cells                                                                        Johan W. Lagerberg

18.       Cord Blood Clinical Processing, Cryopreservation,  and Storage                 

Heidi Elmoazzen and Jelena L. Holovati

19.       Directional Freezing For Large Volume Cryopreservation                                         Joseph Saragusty 

20        Vitrification Of Heart Valve Tissues                                                              Kelvin G.M. Brockbank, Zhenzhen Chen, Elizabeth D. Greene,

and Lia H. Campbell

21        Cryopreservation of Plant Cell Lines                                                              Heinz Martin Schumacher, Martina Westphal, and Elke Heine-Dobbernack

22        Writing Standard Operating Procedures (Sops) For Cryostorage Protocols:

Using Shoot Meristem Cryopreservation As An Example                                         Keith Harding and Erica E. Benson

PART IV: Freeze-Drying Protocols                                                                                   

23        Freeze-Drying Of Proteins                                                                                          Baolin Liu, Xinli Zhou

24        Freeze-Drying Of Lactic Acid Bacteria                                                                     Fernanda Fonseca, Stéphanie Cenard, Stéphanie Passot

25        Freeze-Drying Of Mammalian Sperm                                                                         Levent Keskintepe, Ali Eroglu

26        Freeze-Drying Of Decellularized Heart Valve Tissues                                               Willem F. Wolkers, Andres Hilfiker