Mechanical/Physical Methods of Cell Disruption and Tissue Homogenization
This chapter covers the various methods of mechanical cell disruption and tissue homogenization that are currently commercially available for processing minute samples (<1 ml) to larger production quantities. These mechanical methods of lysing do not introduce chemicals or enzymes to the system. However, the energies needed when using these “harsh” methods can be high and destroy the very proteins being sought.
The destruction of cell membranes and walls is effected by subjecting the cells (1) to shearing by liquid flow, (2) to exploding by pressure differences between inside and outside of cell, (3) to collision forces by impact of beads or paddles, or (4) a combination of these forces. Practical suggestions to optimize each method, where to acquire such equipment, and links to reference sources are included.
Key WordsCell disruption bead mills BioNeb cell disruption cell disruption bomb douce tissue grinder Dyno-mill French press Gaulin high pressure homogenizer high pressure homogenizers Megatron Microfluidics mixer-mill mortar pestle nitrogen Parr bomb opposed jet homogenization Parr nitrogen bomb Polytron Potter-Elvehjem tissue grinders pressure bomb Sonicator Sonitube tissue grinders tissue homogenization ultrasonic processor
Superb assistance was rendered by the following people who are most conversant in their noted equipment areas. Bead milling by Tim Hopkins (BIOSPEC PRODUCTS) and Norbert Roskosch (W. A. BACHOFEN AG); Rotor/Stators by Roger Munsinger (KINEMATICA USA/AG); High-pressure flowing liquid by William Pandolfe (INVESYS) and Thomai Panagiotou (MICROFLUIDICS); High-pressure batch gas by Deanna Shepard (PARR INSTRUMENTS); Low-pressure flowing gas by Lee Clark (GLAS-COL); and ultrasonic processors by Marc Lusting and Andrea Coppola (MISONIX).
My book editor Dr. Anthon Posch has been exceptionally supportive in both technical as well as motivational matters. My wife Robin and two sons Evan and Adam are the sparkle of my life.
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