A range of proposed alkyne Raman tags are examined in-silico for activity and then synthesised generating a library of analogues of the natural product, anisomycin. We report the use of bisaryl butadiyne-anisomycin (BADY-anisomycin) in intracellular SRS microscopy studies of uptake and localisation within live and fixed cells. Following rational design and synthesis, BADY-anisomycin was shown to produce an intense Raman band at 2219 cm<sup>-1</sup>, that is centrally located within the cellular silent region and is approximately 60 times more Raman active than the corresponding propargylanisomycin. Finally, we demonstrate two-colour imaging utilising EdU, an alkyne-containing proliferation probe and BADY-anisomycin.
Wear debris produced from articulating prosthetic joints is thought to be phagocytosed by macrophages which then
release pro-inflammatory cytokines leading to the eventual aseptic loosening of the implant. Currently it is difficult to
image wear particles within cells due to the lack of suitable ways of introducing tag molecules into the materials. We
report how coherent anti-Stokes Raman scattering (CARS) spectroscopy can be used to image unlabeled material within
cells relying on inherent chemical contrast. Using model particles we show how CARS signals change with respect to
size and environment of the scattering particle. Incubating particles of polystyrene, polymethylmethacrylate and
polyethylene with RAW264.7 macrophage cells, we demonstrate that it is possible to image cells phagocyotosing
particles as well as to characterize the location of particles in three dimensions using the inherent optical sectioning
ability of CARS. These results suggest that CARS provides an important tool for monitoring the accumulation of wear
debris generated from prosthetic implants.