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Modified SEM Preparation for THP-1 Monocytes

Heriberto DeLeon III, Anxui Kuang, Francisco Rugama and Thomas Eubanks

THP-1 monocytes are non-adherent white blood cells that are typically differentiated into macrophage like cells to induce cellular adherence for cellular viability assays. However, the induced differentiation also alters the external cellular morphology of THP-1 cells, therefore protocol modification and development has led to a novel method of preparation of THP-1 monocytes for scanning electron microscopy. Incorporation of a polystyrene substrate, modification of the dehydration gradient and removal of critical point drying has been shown effective in preparing THP-1 monocytes for observation without alteration of external structure. Analysis of cellular integrity was subsequently via treatment with 100 nm recombinant Panton-Valentine leukocidin to induce cell lysis and allow observation of internal cellular composition and quality of preservation of the proposed preparatory protocol.