Nonlinear multicontrast microscopy of hematoxylin-and-eosin-stained histological sections.
Imaging hematoxylin-and-eosin-stained cancerous histological sections with multicontrast nonlinear excitation fluorescence, second- and third-harmonic generation (THG) microscopy reveals cellular structures with extremely high image contrast. Absorption and fluorescence spectroscopy together with second hyperpolarizability measurements of the dyes shows that strong THG appears due to neutral hemalum aggregation and is subsequently enhanced by interaction with eosin. Additionally, fluorescence lifetime imaging microscopy reveals eosin fluorescence quenching by hemalums, showing better suitability of only eosin staining for fluorescence microscopy. Multicontrast nonlinear microscopy has the potential to differentiate between cancerous and healthy tissue at a single cell level.
Tuer
Adam
A
University of Toronto, Department of Physics, Institute for Optical Sciences, Department of Chemical and Physical Sciences, 3359 Mississauga Road North, Mississauga, Ontario, L5L 1C6 Canada. adam.tuer@utoronto.ca
Tokarz
Danielle
D
Prent
Nicole
N
Cisek
Richard
R
Alami
Jennifer
J
Dumont
Daniel J
DJ
Bakueva
Ludmila
L
Rowlands
John
J
Barzda
Virginijus
V
D000465:Algorithms; D003287:Contrast Media; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin; D007089:Image Enhancement / Q000379:methods*; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008853:Microscopy / Q000379:methods*; D017711:Nonlinear Dynamics; D010363:Pattern Recognition, Automated / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D003287:Contrast Media; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin