New Approaches to Old Techniques in Cell Handling for Microscopy.

Appropriate concentrations of reagents, an absence of significant cell clumps and debris, minimization of artifacts and ensuring satisfactory cell preservation directly affect the quality of data generated and cannot be overestimated. Traditionally, cells in suspension are prepared using a cytospin, which uses centrifugal force to concentrate and deposit cells onto a glass slide. Adherent cells are traditionally grown on coverslips located on the bottom of the wells of cell culture plates, or using special chamber slide systems. In our laboratory, we developed and tested simplified homemade approaches for both techniques, allowing users to perform large volume cell functional tests followed by microscopy evaluation without a need for a cytospin, special chamber slide systems or the use of round cover slips. We present methods and illustrative examples involving the cellular uptake of self-delivering oligonucleotides in murine splenocytes and in two adherent human tumor cell lines.