Original scientific paper
https://doi.org/10.3325/cmj.2014.55.186
Visualization of melanoma tumor with lectin-conjugated rare-earth doped fluoride nanocrystals
Tetiana Dumych
orcid.org/0000-0002-8489-5600
; Institute of Cell Biology, National Academy of Sciences of Ukraine Lviv, Ukraine
Maxym Lutsyk
; Danylo Halytsky Lviv National Medical University, Lviv, Ukraine
Mateusz Banski
; Wroclaw University of Technology, Institute of Physics, Wroclaw, Poland
Antonina Yashchenko
; Danylo Halytsky Lviv National Medical University, Lviv, Ukraine
Bartlomiej Sojka
; Wroclaw University of Technology, Institute of Physics, Wroclaw, Poland
Rostyslav Horbay
; Institute of Cell Biology, National Academy of Sciences of Ukraine Lviv, Ukraine
Alexander Lutsyk
; Danylo Halytsky Lviv National Medical University, Lviv, Ukraine
Rostyslav Stoika
; Institute of Cell Biology, National Academy of Sciences of Ukraine Lviv, Ukraine
Jan Misiewicz
; Wroclaw University of Technology, Institute of Physics, Wroclaw, Poland
Artur Podhorodecki
; Wroclaw University of Technology, Institute of Physics, Wroclaw, Poland
Rostyslav Bilyy
orcid.org/0000-0002-2344-1349
; Institute of Cell Biology, National Academy of Sciences of Ukraine Lviv, Ukraine
Abstract
Aim To develop specific fluorescent markers for melanoma
tumor visualization, which would provide high selectivity
and reversible binding pattern, by the use of carbohydrate-
recognizing proteins, lectins, combined with the
physical ability for imaging deep in the living tissues by utilizing
red and near infrared fluorescent properties of specific
rare-earth doped nanocrystals (NC).
Methods B10F16 melanoma cells were inoculated to
C57BL/6 mice for inducing experimental melanoma tumor.
Tumors were removed and analyzed by lectin-histochemistry
using LABA, PFA, PNA, HPA, SNA, GNA, and
NPL lectins and stained with hematoxylin and eosin. NPL
lectin was conjugated to fluorescent NaGdF4:Eu3+-COOH
nanoparticles (5 nm) via zero length cross-linking reaction,
and the conjugates were purified from unbound substances
and then used for further visualization of histological
samples. Fluorescent microscopy was used to visualize
NPL-NaGdF4:Eu3+ with the fluorescent emission at 600-720
nm range.
Results NPL lectin selectively recognized regions of undifferentiated
melanoblasts surrounding neoangiogenic foci
inside melanoma tumor, PNA lectin recognized differentiated
melanoblasts, and LCA and WGA were bound to tumor
stroma regions. NPL-NaGdF4:Eu3+ conjugated NC were
efficiently detecting newly formed regions of melanoma
tumor, confirmed by fluorescent microscopy in visible and
near infrared mode. These conjugates possessed high photostability
and were compatible with convenient xylenebased
mounting systems and preserved intensive fluorescent
signal at samples storage for at least 6 months.
Conclusion NPL lectin-NaGdF4:Eu3+ conjugated NC permitted
distinct identification of contours of the melanoma
tissue on histological sections using red excitation at 590-
610 nm and near infrared emission of 700-720 nm. These
data are of potential practical significance for development
of glycans-conjugated nanoparticles to be used for in vivo
visualization of melanoma tumor.
Keywords
Hrčak ID:
127313
URI
Publication date:
15.6.2014.
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