Flow Cytometry and Cell Sorting Laboratory
High Speed Cell Sorter
Dako Cytomation MoFlo
The Cytomation Inc. MoFlo (Modular Flow cytometer) provides our sorting capability. In addition, the MoFlo provides the capabilities for the more complicated experiments that can not be done on the FACScan or FACSCalibur (e.g. more than four colors simultaneously), other fluorochrome emission wavelengths, and requirements for other laser excitations (e.g. ultraviolet). Our MoFlo is configured with 1 iCyte 200mw Blue Sappire 488nm laser and a Melles Groit Helium-Neon laser. The He-Ne produces 35mW of 633nm light. We also have a UV laser that is tunable but normally operates at 350nm. It is possible to tune this laser to the following wavelengths: 350nm, 407nm, 490-530nm, and 568nm. The MoFlo is capable of analyzing up to 10 colors plus forward and side scatter. In terms of sorting capability, the MoFlo can sort for up to four different cell populations at once and is capable of purifying populations that are less than 1% of the total input population. The MoFlo was the first commercial instrument developed with high speed sorting. We have done sorts as high as 50,000cells/sec with recovery rates of 90% or greater. Typical sorts on our MoFlo result in purified populations of 95% or greater. We have the ability to sort a wide variety of cell types due to the variable sizes of our interchangeable nozzle tips. We can sort materials that range in size from DNA to material (e.g. cells, bacteria, yeast) that is up to 150 um in size. The MoFlo can sort into a variety of capture vessels ranging from 1.5ml microcentrifuge tubes to 15ml to 50ml conical centrifuge tubes. In addition to test tubes typically used for bulk sorts, the Cyclone on the MoFlo can deposit variable number of cells into a variety of catch vessels; 6, 12, 24, 48, 96, 384 well cluster plates as well as onto slides. It is also possible to sort cells directly onto filters or nitrocellulose membranes. Individual cells can be sorted into plates or slides for cloning for hybridoma production or PCR analysis or in-situ analysis.