Sodium chloride in separation medium enhances cell compatibility of free flow electrophoresis.
Free flow electrophoresis of cell suspensions in buffers containing sodium chloride was investigated using a modified procedure and the new apparatus Octopus PZE. The major methodical innovations are upward fluid flow, margin buffers flowing through the electrophoresis chamber at both sides of a central cell suspension buffer, adjacent to the electrode membranes, and a sample injection device which focuses the cells hydrodynamically to the middle of the chamber thickness. Mononuclear leukocytes, suspended in a buffer containing 35 mM NaCl, could be fractionated with the same accuracy as by conventional free flow electrophoresis, operated with a single NaCl-free chamber buffer. However, testing the vitality of separated cells with the help of the calcium indicator FURA2-AM clearly demonstrated the biological importance of the presence of a minimum amount of sodium chloride during cell electrophoresis. Only if at least 35 mM NaCl were present could an undisturbed cytosolic Ca2+ metabolism be maintained for the time of a free flow electrophoresis cell separation experiment.
Bondy
B
B
Neurochemistry Department, Psychiatric Clinic of University Munich, Germany.
Bauer
J
J
Seuffert
I
I
Weber
G
G
D001402:B-Lymphocytes / Q000502:physiology; D002021:Buffers; D002118:Calcium / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D004396:Coloring Agents; D004586:Electrophoresis / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry; D012965:Sodium Chloride; D013601:T-Lymphocytes / Q000502:physiology
D002021:Buffers; D004396:Coloring Agents; D012965:Sodium Chloride; D002118:Calcium