Answer

What can I do if my cell numbers are very low?

For optimal results, T cells and antigen-presenting cells need to form a monolayer on the membrane. If T cells are limiting (e.g. when testing cells isolated form a solid organ or biopsy material) any number of T cells can be plated on a monolayer of APCs. If both cell types are limiting, CTL recommends using the low-volume plates manufactured by Whatman. Alternatively, the activation culture can be done in regular U- or V-bottom plates, followed by a transfer of the cells into the ELISPOT plate. (Pipet very gently to minimize shear forces during the transfer.)

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