Perfusion systems: continuous medium flow for cell culture

Perfusion systems

Long-term imaging of cultured cells requires controlled environmental conditions. Continuous perfusion of the medium allows maintaining cells under optimal physiological conditions, with constant nutrient supply and controlled pH environment while avoiding cell debris accumulation.

Medium flow in a continuous way and generating minimal shear stress for cells can be achieved through perfusion systems, which allow a flowing medium through positive pressure (pressure controllers) or through negative pressure (vacuum through syringe pumps). Additional components in a perfusion system allow to ensure a highly controlled flow rate through flow sensors, and the possibility to perform fast-medium switches (for instance, when sequential drug injection is required).

To be maintained under continuous medium flow, cells can be cultured in commercially available flow chambers, commercially available microfluidic devices, or homemade chambers. Perfusion systems can be coupled to temperature controllers for a fully controlled environment during live-cell imaging.

R. Li, The Art of Choreographing Asymmetric Cell Division, Dev cell, 2013

LS. Rose  P. Gönczy Asymmetric cell division and axis formation in the embryo, worm book 2013

NW Goehring, PK Trong, JS Bois,  D Chowdhury, EM Nicola, AA Hyman, Grill SW.Polarization of PAR proteins by advective triggering of a pattern-forming system, Science, 2011

CR.Cowan, AA. Hyman. Asymmetric cell division in C. elegans: cortical polarity and spindle positioning. Annu Rev Cell Dev Biol. 2004.

KF. O’Connell, CM. Leys and JG. White A Genetic Screen for Temperature-Sensitive Cell-Division Mutants of Caenorhabditis elegans genetics 1998

Tsou MF, Ku W, Hayashi A, Rose LS. PAR-dependent and geometry-dependent mechanisms of spindle positioning. J Cell Biol. 2003

S. Ben-Aroya, X. Pan, JD. Boeke, and P. Hieter, Making temperature-sensitive mutants, Methods Enzymol. 2010