Cryobiology - A Short Course

Locksley E. McGann, Dept. of Pathology,
University of Alberta, Alberta, Canada.

This web-based textbook is intended as an introduction to the science of cryobiology-- the study of living things at low temperatures. It is a work in progress and will be built up piecemeal, as time permits. You may find various sections assume wildly different backgrounds for the reader, but hopefully as work progresses, this will become less of a problem.

Note: You will need IBM's techexplorer plugin to view the equations in this document as they are formatted in TeX. Go to IBM's site to get it.

Contents

1. Life at Low Temperatures

• What is Temperature?
• What is Life?
  • Where Does Life Come From?
  • The Fundamental Unit of Life
  • What do Living Things Do?
  • What Makes an Organism?
  • Communication Between Cells
  • Cells That Form a Communication Channel
  • Colligative Properties of Life

2. Historical Overview

• Introduction
• Approach to this Course
• Historical Perspective

3. Water

• Structure and Properties of Water
  • The Water Molecule
  • Water Structure
  • Models of Liquid Water
  • Properties of Water
  • Polywater
• Cell Water
  • Opposing concepts in Cell Physiology
  • Historical Development
  • Emergence of the Membrane Pump Theory
  • Modern Bulk Phase Theories

4. Aqueous Solutions

• Aqueous Solutions - (PDF file)
  • Solutions
  • Concentration
  • Solubility
  • Colligative Properties
  • Vapor Pressure and Raoult's Law
  • Freezing Point Depression
  • Boiling Point Elevation
• Osmotic Pressure - (PDF file)
  • Thermodynamics
  • Kinetic Theory
  • Intermolecular Forces
  • Thermodynamic Relation
  • Non-Ideal Solutions
  • The Osmotic Coefficient
  • Osmoles
  • Osmosis in Cells
  • The Boyle-Van't Hoff Relation

5. Permeability

• Diffusion - (PDF file)
  • Molecular Motion
  • Diffusion
• Cell Permeability - (PDF file)
  • Membrane Structure
  • Semi-Permeable Membranes
  • Coupled Transport of Solvent and Solute

6. Physical Aspects of Freezing

• Phase Diagrams - (PDF file)
  • Mixtures of Phases of Matter
  • Binary Phase Diagrams
  • Ternary Phase Diagrams
  • Glycerol-NaCl-Water
  • DMSO-NaCl-Water
• The Freezing Process - (PDF file)
  • Formation of Ice
  • Nucleation of Ice
    • Thermodynamic Considerations
      • Derivation of Critical Radius
      • Heterogeneous Nucleation
  • Growth of Ice Crystals
    • Ice Structure
      • The Water Molecule
      • Latent Heat of Fusion
    • Dynamics of Crystal Growth
      • Cellular Ice Growth
      • Dendritic Breakdown
      • Recrystallization
    • Solidification Without Crystallization

7. Freezing of Cells

• Investigations into Freeze Thaw Injury
  • Investigations into Cryoinjury
  • Hypotheses of the Mechanisms of Injury During Freezing and Thawing
  • Investigations into the Mechanism of Intracellular Freezing Injury
• Synthesis: Tentative Models of Cryoinjury
  • Slow Cooling Injury - Post-Hypertonic Lysis
    • A Mechanism for Post-Hypertonic Lysis
    • Post-Hypertonic Lysis - Model Details
    • Post-Hypertonic Lysis - The Model in Action
    • Post-Hypertonic Lysis - Reality Check
  • Rapid Cooling Injury - Osmotic Poration

8. Cryopreservation of Cells

• Cryoprotectants
  • Cryoprotection
  • Graded Freezing
  • Penetrating Cryoprotectants
  • Non-Penetrating Cryoprotectants
  • Mechanisms of Cryoprotection - Penetrating Agents
  • Mechanisms of Cryoprotection - Non-Penetrating Agents
• Viability Assays
  • What is Life? What is Viability?
  • Types of Viability Assays
  • Validity of Viability Assays
  • Classification of Viability Assays
• Procedures for Cryopreserving Cells
  • Preparation of Cells
  • Freezing and Storage Vessels
  • Freezing, Storage, and Thawing
  • Safety

9. Cryopreservation of Tissues and Organs

• Cryopreservation of Tissues
  • Tissue Architecture
  • Penetration of Cryoprotectants
  • IIF propagation
  • Mechanical Damage from Freezing
    • Planar vs. Dendritic Ice Growth
    • Ice Growth in Porous Media
    • Ice Growth in Articular Cartilage
    • Vascular Damage
  • Tissue Banking
  • A Short Catalog of Cryopreservation Protocols
  • Quality Control in Tissue Banking
• Cryopreservation of Organs
  • Organ Transplantation
  • Approaches to Organ Preservation
    • Hypothermic Injury and Storage
    • Freezing and Thawing
    • Vitrification

10. Cryosurgery

• Cancer
• Cryoinjury
  • Cellular Injury
  • Secondary Biological Injury
  • The Cryosurgical Lesion
• Factors Affecting Cryosurgical Injury
  • Cooling Rate
  • Tissue Temperature
  • Time vs. Temperature
  • Warming Rate
  • Repetition of the Freeze-Thaw Cycle
• Optimization of Clinical Cryosurgery
• Tissue Cryo-Engineering

11. Hypothermia and Frostbite

• Temperature Control in Mammals
  • Heat Loss
  • Heat Production
  • Physiological Response to Cooling
• Hypothermia
  • Injury
  • Survival and Treatment
  • Medical Uses
  • A Cryobiologist in the Cold
• Frostbite
  • Injury
  • Treatment
  • A Cryobiologist gets even Colder
• Trenchfoot, Paddyfoot, and the Chilblains

12. Strategies for Survival in Cold Climates

• Mammalian Hibernation
  • Sleep and Torpor
  • Hibernation
    • Onset of Torpor
    • Physiology During Torpor
    • Arousal from Torpor
  • Supercooling
• Freeze Tolerant Animals
  • Freeze Tolerance in Amphibians and Reptiles
    • Assessment and Control of Ice Formation
    • Cryoprotection
    • Lowered Metabolism and Limiting of Ischemia
    • Reanimation
  • Aquatic Animals at Low Temperatures
    • Antifreeze Proteins
  • Insect Adaptations to Low Temperatures
    • Freeze Tolerance
      • Cryoprotectants
      • Ice Nucleating Proteins
      • Stabilization of Bound Water
      • Antifreeze Proteins
      • Vitrification
    • Freeze Avoidance
      • Colligative Freezing Point Depression
      • Non-Colligative Freezing Point Depression
      • Supercooling
• Plants at Low Temperatures
  • Chilling Injury
  • Cold Adaptation and Freeze Avoidance

13. Food Preservation by Freezing

• Freeze-Preservation of FoodStuffs
• Freezing of Plants
• Freezing of Meats

14. Freezing Technology

• Cryogenics
  • Refrigeration
    • Pressure Change Cooling
    • Phase Change Cooling
    • The Vapor-Compression Refrigeration Cycle
    • Thermoelectric Cooling
    • Vortex Cooling
    • Thermoacoustic Cooling
    • Aside: The Einstein-Szilard Refrigeration Company
  • Cryogenic Liquifaction
• Temperature Measurement
  • Expansion-Contraction Thermometers
  • Resistance Thermometers
  • Thermocouples
  • Thermistors
  • Semi-Conductor Thermometers
  • Infrared Temperature Measurement
  • Comparison of Methods of Thermometry
• Cryomicroscopy
  • Historical Development
  • Convection Cryostage
  • Conduction Cryostage
  • Directional Solidification Cryostage
  • Common Problems encountered doing Cryomicroscopy

15. Cryonics

• Time travel
• Resurrection (a leap of faith)
• What is Death?
• Will the Future Want Us?
• Current procedures
• What science exists?
• Matters Arising