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Croatian medical journal, Vol.55 No.3 June 2014. -

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.

Autor: Tetiana Dumych - ; Institute of Cell Biology, National Academy of Sciences of Ukraine Lviv, Ukraine



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