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Optica Publishing Group
  • Applied Spectroscopy
  • Vol. 63,
  • Issue 10,
  • pp. 1181-1186
  • (2009)

Demountable Liquid/Flow Cell for in Vivo Infrared Microspectroscopy of Biological Specimens

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Abstract

We have developed a liquid/flow cell/chamber allowing infrared measurements of living biological specimens with high spatial resolution under a controlled aqueous environment. This flow chamber features sub-micrometer thick diamond windows exhibiting low spherical and chromatic aberrations. Diamond has excellent transmission properties and minimal dispersion over the entire mid-infrared and visible spectral ranges. In contrast to current commercially available infrared liquid chambers, the flow chamber has a slim profile, which accommodates high resolution/magnification microscope objectives with small working distances, down to 0.6 mm above the chamber and 6 mm below the flow chamber. We have coupled a pump to the flow chamber to provide medium exchange. As an example, we present microspectroscopic infrared maps and spectra of the freshwater green alga <i>Micrasterias</i> sp. in the new flow chamber and compare them to maps obtained with a conventional liquid chamber. Pulse-amplitude-modulated fluorescence measurements on <i>Micrasterias</i> sp. cells inside the new flow chamber have been evaluated to demonstrate the viability of the algal cells.

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