Non-invasive live-cell measurement of changes in macrophage NAD(P)H by two-photon microscopy.
Two-photon microscopy allows determination of UV-excitable fluorophores using long-wavelength light. We aimed to determine NAD(P)H autofluorescence as a measure for macrophage NADPH-oxidase activation. RAW264.7 macrophages were grown on glass coverslips and kept in HBSS for microscopic investigation. Cells were excited with 710 nm light and NAD(P)H autofluorescence was detected. Glucose as well as NaCN evoked an increase of NAD(P)H autofluorescence. Activators of NADPH oxidase lead to significantly decreased NAD(P)H autofluorescence. Therefore, this work shows the suitability of two-photon microscopy as a non-invasive method determining changes in phagocyte NAD(P)H upon activation.
Kable
Eleanor P W
EP
Electron Microscope Unit, Australian Key Centre for Microscopy & Microanalysis, Madsen Building FO9, University of Sydney, NSW 2006, Sydney, Australia.
Kiemer
Alexandra K
AK
D000818:Animals; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D009243:NAD / Q000378:metabolism*; D009249:NADP / Q000378:metabolism*; D019255:NADPH Oxidases / Q000378:metabolism
D009243:NAD; D009249:NADP; D019255:NADPH Oxidases