Detection of apoptosis using fluorescent probes.

Apoptosis plays a major role in both healthy and diseased cells. The analysis of apoptosis can take advantage of multiple cellular markers, enabling the process to be studied at different time points. In this chapter, several apoptosis assay protocols are provided.

Synthesis of a red fluorescent dye-conjugated Ag@SiO2 nanocomposite for cell immunofluorescence.

In this work we describe a one-step approach for incorporating a red fluorophore (2SBPO) into core-shell nanoparticles for metal-enhanced fluorescence immunolabels. The 2SBPO-MEF nanoparticles are particularly attractive as cell labels because their ∼ 670 nm emission has minimal overlap with cell autofluorescence and from overlap with many conventional probes. 2SBPO was incorporated through physical entrapment during the Stöber process.

Facile Functionalization of Ag@SiO Core-Shell Metal Enhanced Fluorescence Nanoparticles for Cell Labeling.

We describe a versatile approach for functionalizing core-shell Ag@SiO nanoparticles for live-cell imaging. The approach uses physical adsorption and does not need covalent linkage to synthesize antibody-based labels. The surface orientation is not controlled in this approach, but the signal enhancement is strong and consistent.

Single molecule fluorescence correlation spectroscopy of single apoptotic cells using a red-fluorescent caspase probe.

The detection of single molecules in single cells has enabled biochemical analyses to be conducted with high sensitivity and high temporal resolution. In this work, detection of apoptosis was studied by single molecule fluorescence correlation spectroscopy (FCS) in single living cells. Caspase activity was assayed using a new red fluorogenic probe that avoids the spectral overlap of green fluorescent probes and cell autofluorescence.