Chapter 4.1 General methods

Crystallography of biological macromolecules

Second Online Edition (2012)

Part 4. Crystallization

  1. C. Sauter1,
  2. B. Lorber2,
  3. A. McPherson3,
  4. R. Giegé4

Published Online: 14 APR 2012

DOI: 10.1107/97809553602060000812

International Tables for Crystallography

International Tables for Crystallography

How to Cite

Sauter, C., Lorber, B., McPherson, A. and Giegé, R. 2012. General methods. International Tables for Crystallography. F:4:4.1:99–121.

Author Information

  1. 1

    Institut de Biologie Moléculaire et Cellulaire (IBMC), Centre National pour la Recherche Scientifique (CNRS), 15 rue René Descartes, Strasbourg, F‐67084, France

  2. 2

    UPR 9002, IBMC–CNRS, 15 rue René Descartes Cedex, Strasbourg, 67084, France

  3. 3

    Department of Molecular Biology and Biochemistry, University of California, 560 Steinhaus, Irvine, CA92697–3900, USA

  4. 4

    Machineries Traductionnelles, ARN, UPR 9002, IBMC du CNRS, 15 rue René Descartes, Strasbourg, 67084, France

Publication History

  1. Published Online: 14 APR 2012



The preparation of well diffracting crystals is a crucial step of any crystallographic project. This chapter defines the nature of biomacromolecular crystals, sketches the theoretical background of crystallogenesis and describes the practical approaches currently available for crystal growth in structural biology. Chemical, biochemical and physical parameters that affect crystallization are discussed. Emphasis is given to the importance of purity and of structural homogeneity. The classical crystallization methodologies are described, such as the favoured vapour‐phase equilibration, batch and dialysis techniques, and the less widely used but powerful counter‐diffusion methods, and the new trends for their miniaturization and automation are outlined. Advanced methodologies are also covered such as growth methods in convection‐free media (i.e. in microfluidic devices or in gels). Furthermore, the behaviour of macromolecules during the prenucleation, nucleation and growth steps is discussed from the physics viewpoint, and ways to enhance crystal perfection are given. In a practical perspective, strategies to efficiently prepare crystals for structural biology and to overcome crystallization bottlenecks are proposed.


  • batch methods of crystallization;
  • crystal growth;
  • crystal quality;
  • crystal seeding;
  • crystallization;
  • crystallizing agents;
  • dialysis methods of crystallization;
  • gel acupuncture method;
  • crystallization in gels;
  • interface diffusion method of crystallization;
  • microgravity;
  • nucleation;
  • prenucleation;
  • vapour‐diffusion methods of crystallization