Multispectral LEDs Eliminate Lipofuscin-Associated Autofluorescence for Immunohistochemistry and CD44 Variant Detection by in Situ Hybridization in Aging Human, non-Human Primate, and Murine Brain

A major limitation of mechanistic studies in aging brains is the lack of routine methods to robustly visualize and discriminate the cellular distribution of tissue antigens using fluorescent immunohistochemical multi-labeling techniques. Although such approaches are routine in non-aging brains, they...

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Bibliographic Details
Published in:ASN neuro 2022, Vol.14, p.17590914221123138
Main Authors: Adeniyi, Philip A., Fopiano, Katie-Anne, Banine, Fatima, Garcia, Mariel, Gong, Xi, Keene, C. Dirk, Sherman, Larry S., Bagi, Zsolt, Back, Stephen A.
Format: Article
Language:English
Subjects:
Aging
Aging - genetics
Aging - metabolism
Alzheimer's disease
Animals
Antigens
Brain - metabolism
Brain research
CD44 antigen
Colorimetry
Dementia
Genetic Variation
Humans
Hyaluronan Receptors - genetics
Hybridization
Immunohistochemistry
In Situ Hybridization
Lipofuscin - chemistry
Mice
Original Papers
Pigments
Primates - genetics
Substantia alba
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Summary:A major limitation of mechanistic studies in aging brains is the lack of routine methods to robustly visualize and discriminate the cellular distribution of tissue antigens using fluorescent immunohistochemical multi-labeling techniques. Although such approaches are routine in non-aging brains, they are not consistently feasible in the aging brain due to the progressive accumulation of autofluorescent pigments, particularly lipofuscin, which strongly excite and emit over a broad spectral range. Consequently, aging research has relied upon colorimetric antibody techniques, where discrimination of tissue antigens is often challenging. We report the application of a simple, reproducible, and affordable protocol using multispectral light-emitting diodes (mLEDs) exposure for the reduction/elimination of lipofuscin autofluorescence (LAF) in aging brain tissue from humans, non-human primates, and mice. The mLEDs lamp has a broad spectral range that spans from the UV to infrared range and includes spectra in the violet/blue and orange/red. After photo quenching, the LAF level was markedly reduced when the tissue background fluorescence before and after mLEDs exposure was compared (p 
ISSN:1759-0914
1759-0914
DOI:10.1177/17590914221123138