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Article ; Online: Biophotonics for lymphatic theranostics in animals and humans.

Tuchin, Valery V / Zharov, Vladimir P / Galanzha, Ekaterina I

2018 Volume 11, Issue 8, Page(s) e201811001

MeSH term(s)	Animals ; Diagnostic Techniques and Procedures ; Humans ; Lymphatic System ; Optics and Photonics ; Therapeutics/methods
Language	English
Publishing date	2018-07-05
Publishing country	Germany
Document type	Editorial ; Introductory Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2390063-5
ISSN	1864-0648 ; 1864-063X
ISSN (online)	1864-0648
ISSN	1864-063X
DOI	10.1002/jbio.201811001
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Article: Optical clearing for photoacoustic lympho- and angiography beyond conventional depth limit

Novoselova, Marina V / Abakumova, Tatiana O / Khlebtsov, Boris N / Zatsepin, Timofei S / Lazareva, Ekaterina N / Tuchin, Valery V / Zharov, Vladimir P / Gorin, Dmitry A / Galanzha, Ekaterina I

Photoacoustics

2020 Volume 20, Page(s) 100186

Abstract: Photoacoustic (PA) imaging (PAI) is an emerging powerful tool for noninvasive real-time mapping of blood and lymphatic vessels and lymph ... ...

Abstract	Photoacoustic (PA) imaging (PAI) is an emerging powerful tool for noninvasive real-time mapping of blood and lymphatic vessels and lymph nodes
Language	English
Publishing date	2020-06-17
Publishing country	Germany
Document type	Journal Article
ZDB-ID	2716706-9
ISSN	2213-5979
ISSN	2213-5979
DOI	10.1016/j.pacs.2020.100186
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Article ; Online: Detection of Apoptotic Circulating Tumor Cells Using in vivo Fluorescence Flow Cytometry.

Nolan, Jacqueline / Nedosekin, Dmitry A / Galanzha, Ekaterina I / Zharov, Vladimir P

Cytometry. Part A : the journal of the International Society for Analytical Cytology

2018 Volume 95, Issue 6, Page(s) 664–671

Abstract: Most cancer patients die from metastatic disease as a result of a circulating tumor cell (CTC) spreading from a primary tumor through the blood circulation to distant organs. Many studies have demonstrated the tremendous potential of using CTC counts as ... ...

Abstract	Most cancer patients die from metastatic disease as a result of a circulating tumor cell (CTC) spreading from a primary tumor through the blood circulation to distant organs. Many studies have demonstrated the tremendous potential of using CTC counts as prognostic markers of metastatic development and therapeutic efficacy. However, it is only the viable CTCs capable of surviving in the blood circulation that can create distant metastasis. To date, little progress has been made in understanding what proportion of CTCs is viable and what proportion is in an apoptotic state. Here, we introduce a novel approach toward in situ characterization of CTC apoptosis status using a multicolor in vivo flow cytometry platform with fluorescent detection for the real-time identification and enumeration of such cells directly in blood flow. The proof of concept was demonstrated with two-color fluorescence flow cytometry (FFC) using breast cancer cells MDA-MB-231 expressing green fluorescein protein (GFP), staurosporine as an activator of apoptosis, Annexin-V apoptotic kit with orange dye color, and a mouse model. The future application of this new platform for real-time monitoring of antitumor drug efficiency is discussed. © 2018 International Society for Advancement of Cytometry.
MeSH term(s)	Animals ; Apoptosis/drug effects ; Biomarkers, Tumor/blood ; Blood Vessels/diagnostic imaging ; Blood Vessels/drug effects ; Breast Neoplasms/diagnostic imaging ; Cell Count ; Cell Line, Tumor ; Cell Survival/drug effects ; Female ; Flow Cytometry/methods ; Fluorescence ; Green Fluorescent Proteins/chemistry ; Green Fluorescent Proteins/metabolism ; Humans ; Mice ; Mice, Nude ; Neoplastic Cells, Circulating/chemistry ; Neoplastic Cells, Circulating/metabolism ; Neoplastic Cells, Circulating/pathology ; Neoplastic Cells, Circulating/radiation effects ; Staurosporine/toxicity
Chemical Substances	Biomarkers, Tumor ; Green Fluorescent Proteins (147336-22-9) ; Staurosporine (H88EPA0A3N)
Language	English
Publishing date	2018-12-03
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2099868-5
ISSN	1552-4930 ; 0196-4763 ; 1552-4922
ISSN (online)	1552-4930
ISSN	0196-4763 ; 1552-4922
DOI	10.1002/cyto.a.23642
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Article: Circulating Tumor Cell Detection and Capture by Photoacoustic Flow Cytometry in Vivo and ex Vivo.

Galanzha, Ekaterina I / Zharov, Vladimir P

Cancers

2013 Volume 5, Issue 4, Page(s) 1691–1738

Abstract: Despite progress in detecting circulating tumor cells (CTCs), existing assays still have low sensitivity (1-10 CTC/mL) due to the small volume of blood samples (5-10 mL). Consequently, they can miss up to 103-104 CTCs, resulting in the development of ... ...

Abstract	Despite progress in detecting circulating tumor cells (CTCs), existing assays still have low sensitivity (1-10 CTC/mL) due to the small volume of blood samples (5-10 mL). Consequently, they can miss up to 103-104 CTCs, resulting in the development of barely treatable metastasis. Here we analyze a new concept of in vivo CTC detection with enhanced sensitivity (up to 102-103 times) by the examination of the entire blood volume in vivo (5 L in adults). We focus on in vivo photoacoustic (PA) flow cytometry (PAFC) of CTCs using label-free or targeted detection, photoswitchable nanoparticles with ultrasharp PA resonances, magnetic trapping with fiber-magnetic-PA probes, optical clearance, real-time spectral identification, nonlinear signal amplification, and the integration with PAFC in vitro. We demonstrate PAFC's capability to detect rare leukemia, squamous carcinoma, melanoma, and bulk and stem breast CTCs and its clusters in preclinical animal models in blood, lymph, bone, and cerebrospinal fluid, as well as the release of CTCs from primary tumors triggered by palpation, biopsy or surgery, increasing the risk of metastasis. CTC lifetime as a balance between intravasation and extravasation rates was in the range of 0.5-4 h depending on a CTC metastatic potential. We introduced theranostics of CTCs as an integration of nanobubble-enhanced PA diagnosis, photothermal therapy, and feedback through CTC counting. In vivo data were verified with in vitro PAFC demonstrating a higher sensitivity (1 CTC/40 mL) and throughput (up to 10 mL/min) than conventional assays. Further developments include detection of circulating cancer-associated microparticles, and super-rsesolution PAFC beyond the diffraction and spectral limits.
Language	English
Publishing date	2013-12-10
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527080-1
ISSN	2072-6694
ISSN	2072-6694
DOI	10.3390/cancers5041691
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Article: In Vivo Lymphatic Circulating Tumor Cells and Progression of Metastatic Disease.

Han, Mikyung / Watts, Julia Alex / Jamshidi-Parsian, Azemat / Nadeem, Urooba / Sarimollaoglu, Mustafa / Siegel, Eric R / Zharov, Vladimir P / Galanzha, Ekaterina I

Cancers

2020 Volume 12, Issue 10

Abstract: The dissemination of circulating tumor cells (CTCs) by lymph fluid is one of the key events in the development of tumor metastasis. However, little progress has been made in studying lymphatic CTCs (L-CTCs). Here, we demonstrate the detection of L-CTCs ... ...

Abstract	The dissemination of circulating tumor cells (CTCs) by lymph fluid is one of the key events in the development of tumor metastasis. However, little progress has been made in studying lymphatic CTCs (L-CTCs). Here, we demonstrate the detection of L-CTCs in preclinical mouse models of melanoma and breast cancer using in vivo high-sensitivity photoacoustic and fluorescent flow cytometry. We discovered that L-CTCs are be detected in pre-metastatic disease stage. The smallest primary tumor that shed L-CTCs was measured as 0.094mm×0.094mm, its volume was calculated as 0.0004 mm
Language	English
Publishing date	2020-10-05
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527080-1
ISSN	2072-6694
ISSN	2072-6694
DOI	10.3390/cancers12102866
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Article ; Online: Photoacoustic flow cytometry.

Galanzha, Ekaterina I / Zharov, Vladimir P

Methods (San Diego, Calif.)

2012 Volume 57, Issue 3, Page(s) 280–296

Abstract: Conventional flow cytometry using scattering and fluorescent detection methods has been a fundamental tool of biological discoveries for many years. Invasive extraction of cells from a living organism, however, may lead to changes in cell properties and ... ...

Abstract	Conventional flow cytometry using scattering and fluorescent detection methods has been a fundamental tool of biological discoveries for many years. Invasive extraction of cells from a living organism, however, may lead to changes in cell properties and prevents the long-term study of cells in their native environment. Here, we summarize recent advances of new generation flow cytometry for in vivo noninvasive label-free or targeted detection of cells in blood, lymph, bone, cerebral and plant vasculatures using photoacoustic (PA) detection techniques, multispectral high-pulse-repetition-rate lasers, tunable ultrasharp (up to 0.8 nm) rainbow plasmonic nanoprobes, positive and negative PA contrasts, in vivo magnetic enrichment, time-of-flight cell velocity measurement, PA spectral analysis, and integration of PA, photothermal (PT), fluorescent, and Raman methods. Unique applications of this tool are reviewed with a focus on ultrasensitive detection of normal blood cells at different functional states (e.g., apoptotic and necrotic) and rare abnormal cells including circulating tumor cells (CTCs), cancer stem cells, pathogens, clots, sickle cells as well as pharmokinetics of nanoparticles, dyes, microbubbles and drug nanocarriers. Using this tool we discovered that palpation, biopsy, or surgery can enhance CTC release from primary tumors, increasing the risk of metastasis. The novel fluctuation flow cytometry provided the opportunity for the dynamic study of blood rheology including red blood cell aggregation and clot formation in different medical conditions (e.g., blood disorders, cancer, or surgery). Theranostics, as a combination of PA diagnosis and PT nanobubble-amplified multiplex therapy, was used for eradication of CTCs, purging of infected blood, and thrombolysis of clots using PA guidance to control therapy efficiency. In vivo flow cytometry using a portable fiber-based devices can provide a breakthrough platform for early diagnosis of cancer, infection and cardiovascular disorders with a potential to inhibit, if not prevent, metastasis, sepsis, and strokes or heart attack by well-timed personalized therapy.
MeSH term(s)	Animals ; Blood Cells/pathology ; Cardiovascular Diseases/diagnosis ; Cardiovascular Diseases/pathology ; Flow Cytometry/instrumentation ; Flow Cytometry/methods ; Humans ; Lasers ; Lymph/cytology ; Mice ; Molecular Imaging/methods ; Nanoparticles/ultrastructure ; Neoplasms/diagnosis ; Neoplasms/pathology ; Neoplastic Cells, Circulating/pathology ; Neoplastic Stem Cells/pathology ; Photoacoustic Techniques
Language	English
Publishing date	2012-06-26
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Review
ZDB-ID	1066584-5
ISSN	1095-9130 ; 1046-2023
ISSN (online)	1095-9130
ISSN	1046-2023
DOI	10.1016/j.ymeth.2012.06.009
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Zs.A 3239: Show issues

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Article ; Online: Lymph Liquid Biopsy for Detection of Cancer Stem Cells.

Han, Mikyung / Watts, J Alex / Jamshidi-Parsian, Azemat / Nadeem, Urooba / Siegel, Eric R / Zharov, Vladimir P / Galanzha, Ekaterina I

Cytometry. Part A : the journal of the International Society for Analytical Cytology

2020 Volume 99, Issue 5, Page(s) 496–502

Abstract: Collection of a blood sample defined by the term "blood liquid biopsy" is commonly used to detect diagnostic, prognostic, and therapeutic decision-making markers of metastatic tumors including circulating tumor cells (CTCs). Many tumors also release CTCs ...

Abstract	Collection of a blood sample defined by the term "blood liquid biopsy" is commonly used to detect diagnostic, prognostic, and therapeutic decision-making markers of metastatic tumors including circulating tumor cells (CTCs). Many tumors also release CTCs and other markers into lymph fluid, but the utility of lymphatic markers largely remains unexplored. Here, we introduce lymph liquid biopsy through collection of peripheral (afferent) and central (thoracic duct [TD]) lymph samples and demonstrates its feasibility for detection of stem-like CTCs potentially responsible for metastasis development and tumor relapse. Stemness of lymphatic CTCs (L-CTCs) was determined by spheroid-forming assay in vitro. Simultaneously, we tested blood CTCs by conventional blood liquid biopsy, and monitored the primary tumor size, early metastasis in a sentinel lymph node (SLN) and distant metastasis in lungs. Using a mouse model at early melanoma stage with no distant metastasis, we identified stem-like L-CTCs in lymph samples from afferent lymphatic vessels. Since these vessels transport cells from the primary tumor to SLN, our finding emphasizes the significance of the lymphatic pathway in development of SLN metastasis. Surprisingly, in pre-metastatic disease, stem-like L-CTCs were detected in lymph samples from the TD, which directly empties lymph into blood circulation. This suggests a new contribution of the lymphatic system to initiation of distant metastasis. Integration of lymph and blood liquid biopsies demonstrated that all mice with early melanoma had stem-like CTCs in at least one of three samples (afferent lymph, TD lymph, and blood). At the stage of distant metastasis, spheroid-forming L-CTCs were detected in TD lymph, but not in afferent lymph. Altogether, our results demonstrated that lymph liquid biopsy and testing L-CTCs holds promise for diagnosis and prognosis of early metastasis. © 2020 International Society for Advancement of Cytometry.
MeSH term(s)	Humans ; Liquid Biopsy ; Lymph Nodes ; Lymphatic Metastasis ; Neoplasm Recurrence, Local ; Neoplastic Cells, Circulating ; Neoplastic Stem Cells ; Sentinel Lymph Node Biopsy
Language	English
Publishing date	2020-09-18
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2099868-5
ISSN	1552-4930 ; 0196-4763 ; 1552-4922
ISSN (online)	1552-4930
ISSN	0196-4763 ; 1552-4922
DOI	10.1002/cyto.a.24221
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Article ; Online: In vivo photoacoustic and photothermal cytometry for monitoring multiple blood rheology parameters.

Galanzha, Ekaterina I / Zharov, Vladimir P

Cytometry. Part A : the journal of the International Society for Analytical Cytology

2011 Volume 79, Issue 10, Page(s) 746–757

Abstract: Alterations of blood rheology (hemorheology) are important for the early diagnosis, prognosis, and prevention of many diseases, including myocardial infarction, stroke, sickle cell anemia, thromboembolism, trauma, inflammation, and malignancy. However, ... ...

Abstract	Alterations of blood rheology (hemorheology) are important for the early diagnosis, prognosis, and prevention of many diseases, including myocardial infarction, stroke, sickle cell anemia, thromboembolism, trauma, inflammation, and malignancy. However, real-time in vivo assessment of multiple hemorheological parameters over long periods of time has not been reported. Here, we review the capabilities of label-free photoacoustic (PA) and photothermal (PT) flow cytometry for dynamic monitoring of hemorhelogical parameters in vivo which we refer to as photoacoustic and photothermal blood rheology. Using phenomenological models, we analyze correlations between both PT and PA signal characteristics in the dynamic modes and following determinants of blood rheology: red blood cell (RBC) aggregation, deformability, shape (e.g., as in sickle cells), intracellular hemoglobin distribution, individual cell velocity, hematocrit, and likely shear rate. We present ex vivo and in vivo experimental verifications involving high-speed PT imaging of RBCs, identification of sickle cells in a mouse model of human sickle cell disease and in vivo monitoring of complex hemorheological changes (e.g., RBC deformability, hematocrit and RBC aggregation). The multi-parameter platform that integrates PT, PA, and conventional optical techniques has potential for translation to clinical applications using safe, portable, laser-based medical devices for point-of-care screening of disease progression and therapy efficiency.
MeSH term(s)	Anemia, Sickle Cell/diagnosis ; Anemia, Sickle Cell/pathology ; Animals ; Blood Viscosity ; Cardiovascular Diseases/diagnosis ; Cardiovascular Diseases/pathology ; Early Diagnosis ; Erythrocyte Aggregation ; Erythrocyte Deformability ; Flow Cytometry/instrumentation ; Flow Cytometry/methods ; Fluorescence ; Hematocrit ; Hemoglobins ; Hemorheology ; Humans ; Lasers ; Mice ; Molecular Imaging/instrumentation ; Molecular Imaging/methods ; Photoacoustic Techniques/instrumentation ; Photoacoustic Techniques/methods ; Signal Processing, Computer-Assisted
Chemical Substances	Hemoglobins
Language	English
Publishing date	2011-08-30
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Review
ZDB-ID	2099868-5
ISSN	1552-4930 ; 0196-4763 ; 1552-4922
ISSN (online)	1552-4930
ISSN	0196-4763 ; 1552-4922
DOI	10.1002/cyto.a.21133
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Article: Indocyanine green dye based bimodal contrast agent tested by photoacoustic/fluorescence tomography setup.

Mokrousov, Maksim D / Thompson, Weylan / Ermilov, Sergey A / Abakumova, Tatiana / Novoselova, Marina V / Inozemtseva, Olga A / Zatsepin, Timofei S / Zharov, Vladimir P / Galanzha, Ekaterina I / Gorin, Dmitry A

Biomedical optics express

2021 Volume 12, Issue 6, Page(s) 3181–3195

Abstract: Multimodal imaging systems are in high demand for preclinical research, experimental medicine, and clinical practice. Combinations of photoacoustic technology with other modalities including fluorescence, ultrasound, MRI, OCT have been already applied in ...

Abstract	Multimodal imaging systems are in high demand for preclinical research, experimental medicine, and clinical practice. Combinations of photoacoustic technology with other modalities including fluorescence, ultrasound, MRI, OCT have been already applied in feasibility studies. Nevertheless, only the combination of photoacoustics with ultrasound in a single setup is commercially available now. A combination of photoacoustics and fluorescence is another compelling approach because those two modalities naturally complement each other. Here, we presented a bimodal contrast agent based on the indocyanine green dye (ICG) as a single signalling compound embedded in the biocompatible and biodegradable polymer shell. We demonstrate its remarkable characteristics by imaging using a commercial photoacoustic/fluorescence tomography system (TriTom, PhotoSound Technologies). It was shown that photoacoustic signal of the particles depends on the amount of dye loaded into the shell, while fluorescence signal depends on the total amount of dye per particle. For the first time to our knowledge, a commercial bimodal photoacoustic/fluorescence setup was used for characterization of ICG doped polymer particles. Additionally, we conducted cell toxicity studies for these particles as well as studied biodistribution over time
Language	English
Publishing date	2021-05-07
Publishing country	United States
Document type	Journal Article
ZDB-ID	2572216-5
ISSN	2156-7085
ISSN	2156-7085
DOI	10.1364/BOE.419461
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Article ; Online: Current status, pitfalls and future directions in the diagnosis and therapy of lymphatic malformation.

Sun, Ravi W / Tuchin, Valery V / Zharov, Vladimir P / Galanzha, Ekaterina I / Richter, Gresham T

Journal of biophotonics

2017 Volume 11, Issue 8, Page(s) e201700124

Abstract: Lymphatic malformations are complex congenital vascular lesions composed of dilated, abnormal lymphatic channels of varying size that can result in significant esthetic and physical impairment due to relentless growth. Lymphatic malformations comprised ... ...

Abstract	Lymphatic malformations are complex congenital vascular lesions composed of dilated, abnormal lymphatic channels of varying size that can result in significant esthetic and physical impairment due to relentless growth. Lymphatic malformations comprised of micro-lymphatic channels (microcystic) integrate and infiltrate normal soft tissue, leading to a locally invasive mass. Ultrasonography and magnetic resonance imaging assist in the diagnosis but are unable to detect microvasculature present in microcystic lymphatic malformations. In this review, we examine existing tools and elaborate on alternative diagnostic methods in assessing lymphatic malformations. In particular, photoacoustics, low-toxicity nanoparticles and optical clearing can overcome existing challenges in the examination of lymphatic channels in vivo. In combination with photothermal scanning and flow cytometry, Photoacoustic techniques may provide a versatile tool for lymphatic-related clinical applications, potentially leading to a single diagnostic and therapeutic platform to overcome limitations in current imaging techniques and permit targeted theranostics of microcystic lymphatic malformations.
MeSH term(s)	Diagnostic Imaging ; Humans ; Lymphatic Abnormalities/diagnosis ; Lymphatic Abnormalities/therapy ; Nanomedicine ; Optical Phenomena
Language	English
Publishing date	2017-11-05
Publishing country	Germany
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. ; Review
ZDB-ID	2390063-5
ISSN	1864-0648 ; 1864-063X
ISSN (online)	1864-0648
ISSN	1864-063X
DOI	10.1002/jbio.201700124
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