Cost-effective elimination of lipofuscin fluorescence from formalin-fixed brain tissue by white phosphor light emitting diode array.
Autofluorescence of aldehyde-fixed tissues greatly hinders fluorescence microscopy. In particular, lipofuscin, an autofluorescent component of aged brain tissue, complicates fluorescence imaging of tissue in neurodegenerative diseases. Background and lipofuscin fluorescence can be reduced by greater than 90% through photobleaching using white phosphor light emitting diode arrays prior to treatment with fluorescent probes. We compared the effect of photobleaching versus established chemical quenchers on the quality of fluorescent staining in formalin-fixed brain tissue of frontotemporal dementia with tau-positive inclusions. Unlike chemical quenchers, which reduced fluorescent probe signals as well as background, photobleaching treatment had no effect on probe fluorescence intensity while it effectively reduced background and lipofuscin fluorescence. The advantages and versatility of photobleaching over established methods are discussed.
a Department of Medical Biophysics, Princess Margaret Cancer Centre, University of Toronto, Toronto, ON M5G 1L7, Canada.
Yulong
Y
Sun
a Department of Medical Biophysics, Princess Margaret Cancer Centre, University of Toronto, Toronto, ON M5G 1L7, Canada.|b Department of Biochemistry, Princess Margaret Cancer Centre, University of Toronto, Toronto, ON M5G 1L7, Canada.
Avi
A
Chakrabartty
D001921:Brain / Q000473:pathology*; D003362:Cost-Benefit Analysis; D005453:Fluorescence; D005455:Fluorescent Antibody Technique; D005557:Formaldehyde / Q000737:chemistry*; D006801:Humans; D008029:Lighting / Q000295:instrumentation*; D008062:Lipofuscin / Q000737:chemistry*; D008856:Microscopy, Fluorescence / Q000191:economics* / Q000379:methods*; D016612:Paraffin Embedding; D038761:Photobleaching / Q000528:radiation effects*; D012666:Semiconductors
D008062:Lipofuscin; D005557:Formaldehyde
autofluorescence; diode électroluminescente; fluorescence microcopy; light emitting diode; lipofuscin; lipofuscine; microscopie en fluorescence; photoblanchiment; photobleaching