Multiphoton Deep-Tissue Imaging of Micrometastases and Disseminated Cancer Cells Using Conjugates of Quantum Dots and Single-Domain Antibodies.
Cell Line, Tumor
Fluorescent Antibody Technique
/ methods
Humans
Immunoconjugates
/ chemistry
Immunohistochemistry
/ methods
Microscopy, Fluorescence, Multiphoton
/ methods
Molecular Probes
Multimodal Imaging
/ methods
Nanoparticles
Neoplasm Micrometastasis
Neoplasms
/ diagnostic imaging
Optical Imaging
/ methods
Quantum Dots
Single-Domain Antibodies
Deep tissue imaging
Disseminated tumor cells
Micrometastases
Multiphoton imaging
Quantum dots
Single-domain antibodies
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
31
7
2021
pubmed:
1
8
2021
medline:
2
9
2021
Statut:
ppublish
Résumé
Early detection of malignant tumors, micrometastases, and disseminated tumor cells is one of the effective way of fighting cancer. Among the many existing imaging methods like computed tomography (CT), ultrasound (US), magnetic resonance imaging (MRI), positron emission tomography (PET), and single-photon emission computed tomography (SPECT), optical imaging with fluorescent probes is one of the most promising alternatives because it is fast, inexpensive, safe, sensitive, and specific. However, traditional fluorescent probes, based on organic fluorescent dyes, suffer from the low signal-to-noise ratio. Furthermore, conventional organic fluorescent dyes are unsuitable for deep tissue imaging because of the strong visible light absorption by biological tissues. The use of fluorescent semiconductor nanocrystals, or quantum dots (QDs), may overcome this limitation due to their large multiphoton cross section, which ensures efficient imaging of thick tissue sections inaccessible with conventional fluorescent probes. Moreover, the lower photobleaching and higher brightness of fluorescence signals from QDs ensures a much better discrimination of positive signals from the background. The use of fluorescent nanoprobes based on QDs conjugated to uniformly oriented high-affinity single-domain antibodies (sdAbs) may significantly increase the sensitivity and specificity due to better recognition of analytes and deeper penetration into tissues due to small size of such nanoprobes.Here, we describe a protocol for the fabrication of nanoprobes based on sdAbs and QDs, preparation of experimental xenograft mouse models for quality control, and multiphoton imaging of deep-tissue solid tumors, micrometastases, and disseminated tumor cells.
Identifiants
pubmed: 34331282
doi: 10.1007/978-1-0716-1593-5_8
doi:
Substances chimiques
Immunoconjugates
0
Molecular Probes
0
Single-Domain Antibodies
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
105-123Informations de copyright
© 2021. Springer Science+Business Media, LLC, part of Springer Nature.
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