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1.
Chem Commun (Camb) ; 56(7): 1093-1096, 2020 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-31894764

RESUMO

We prepared novel conjugated polymer based NIR-II nanoparticles, which display extremely high photothermal conversion efficiency (65%). Both in vitro and in vivo investigations revealed that the as-prepared nanoparticles exhibit excellent theranostic properties including an extremely high cancer cell killing ability, admirable tumor elimination efficiency (100%) and a remarkable photoacoustic imaging contrast enhancing ability.


Assuntos
Antineoplásicos/uso terapêutico , Nanopartículas/uso terapêutico , Compostos de Organossilício/uso terapêutico , Polímeros/uso terapêutico , Tiadiazóis/uso terapêutico , Animais , Antineoplásicos/química , Antineoplásicos/efeitos da radiação , Células Hep G2 , Humanos , Hipertermia Induzida/métodos , Raios Infravermelhos , Camundongos , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Nanopartículas/química , Nanopartículas/efeitos da radiação , Compostos de Organossilício/química , Compostos de Organossilício/efeitos da radiação , Técnicas Fotoacústicas/métodos , Polímeros/química , Polímeros/efeitos da radiação , Nanomedicina Teranóstica/métodos , Tiadiazóis/química , Tiadiazóis/efeitos da radiação
2.
Chem Soc Rev ; 49(1): 21-31, 2020 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-31799523

RESUMO

The results of numerous studies have led to the development of supramolecular (assembled) organic substances for use in biomedical imaging as part of comprehensive approaches to the diagnosis of diseases. This review summarizes recent advances that have been made in the design and fabrication of assembled organic dyes for fluorescence and photoacoustic bioimaging.


Assuntos
Fluorescência , Corantes Fluorescentes/química , Neoplasias/diagnóstico por imagem , Compostos Orgânicos/química , Técnicas Fotoacústicas , Trombose/diagnóstico por imagem , Animais , Corantes Fluorescentes/síntese química , Humanos , Compostos Orgânicos/síntese química
3.
J Surg Oncol ; 121(1): 48-50, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31165483

RESUMO

BACKGROUND AND OBJECTIVES: Photoacoustic lymphangiography, which is based on photoacoustic technology, is an optical imaging that visualizes the distribution of light absorbing tissue components like hemoglobin or melanin, as well as optical absorption contrast imaging agents like indocyanine green (ICG) in the lymphatic channels, with high spatial resolution. In this report, we introduce the three-dimensional (3D) images of human lymphatic vessels obtained with photoacoustic lymphangiography. METHODS: We used the 3D photoacoustic visualization system (PAI-05). Some healthy subjects and lymphedema patients were recruited. To image the lymphatic structures of the limbs ICG was administered subcutaneously as in fluorescence lymphangiography. Photoacoustic images were acquired by irradiating the tissue using a laser at wavelengths of near-infrared region. On the same occasion, fluorescence images were also recorded. RESULTS: The lymphatic vessels up to the diameter of 0.2 mm could be observed three-dimensionally with the venules around them. In the patient-group, dermal backflow patterns were often observed as dense interconnecting 3D structures of lymphatic vessels. Collecting vessels passing below the dermis were also observed, which were not observed by fluorescence lymphography. CONCLUSIONS: Photoacoustic lymphangiography provided the detailed observation of each lymphatic vessel, leading to deeper understanding of 3D structures and physiological state of the vessel.


Assuntos
Vasos Linfáticos/diagnóstico por imagem , Linfedema/diagnóstico por imagem , Linfografia/métodos , Técnicas Fotoacústicas/métodos , Estudos de Casos e Controles , Feminino , Angiofluoresceinografia/métodos , Humanos , Imagem Tridimensional/métodos , Pessoa de Meia-Idade
4.
Nat Med ; 25(12): 1905-1915, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31792454

RESUMO

Biomarkers for monitoring of disease progression and response to therapy are lacking for muscle diseases such as Duchenne muscular dystrophy. Noninvasive in vivo molecular imaging with multispectral optoacoustic tomography (MSOT) uses pulsed laser light to induce acoustic pressure waves, enabling the visualization of endogenous chromophores. Here we describe an application of MSOT, in which illumination in the near- and extended near-infrared ranges from 680-1,100 nm enables the visualization and quantification of collagen content. We first demonstrated the feasibility of this approach to noninvasive quantification of tissue fibrosis in longitudinal studies in a large-animal Duchenne muscular dystrophy model in pigs, and then applied this approach to pediatric patients. MSOT-derived collagen content measurements in skeletal muscle were highly correlated to the functional status of the patients and provided additional information on molecular features as compared to magnetic resonance imaging. This study highlights the potential of MSOT imaging as a noninvasive, age-independent biomarker for the implementation and monitoring of newly developed therapies in muscular diseases.


Assuntos
Colágeno/isolamento & purificação , Imagem Molecular/métodos , Distrofia Muscular de Duchenne/diagnóstico , Tomografia , Animais , Biomarcadores/metabolismo , Criança , Pré-Escolar , Colágeno/classificação , Colágeno/metabolismo , Fibrose/diagnóstico , Fibrose/metabolismo , Fibrose/patologia , Humanos , Masculino , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Distrofia Muscular de Duchenne/metabolismo , Distrofia Muscular de Duchenne/patologia , Técnicas Fotoacústicas , Suínos
5.
Chem Commun (Camb) ; 55(95): 14255-14258, 2019 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-31657388
6.
Int J Nanomedicine ; 14: 7155-7171, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31564871

RESUMO

Background: Platelet activation and subsequent aggregation are the initial stages of thrombosis. A molecular probe that specifically targets activated platelets and remains retained under high shear stress in vivo can enhance the imaging effect to achieve early and accurate diagnosis. Methods and materials: In this study, we constructed nanoparticles (NPs) using polydopamine to carry two peptides that simultaneously bind integrin αIIbß3 and P-selectin on activated platelets to enhance the targeting of NPs to thrombus. Results: The targeting specificity and binding stability of the NPs on red and white thrombi were demonstrated in vitro using a simulated circulatory device and the targeting effect of the NPs on mixed thrombus was studied by  magnetic resonance (MR)/photoacoustic (PA) dual-modality imaging in vivo. NPs that were surface modified with both peptides have higher selectivity and retention to red and white thrombi in vitro than NPs with a single or no peptide, and the targeting effect was closely related to the number and distribution of activated platelets as well as the structure and type of thrombus. The NPs also have MR/PA dual-modality imaging functionality, significantly enhancing the imaging of mixed thrombus in vivo. Conclusion: These dual-targeted NPs have improved targeting specificity and binding stability to different thrombi under high shear stress and are beneficial for the early diagnosis of thrombosis.


Assuntos
Indóis/química , Imagem por Ressonância Magnética , Nanopartículas/química , Técnicas Fotoacústicas , Polímeros/química , Trombose/diagnóstico por imagem , Animais , Meios de Contraste/química , Humanos , Ligantes , Espectroscopia de Ressonância Magnética , Imagem Molecular , Nanopartículas/ultraestrutura , Ativação Plaquetária , Ratos Sprague-Dawley
7.
Cancer Res ; 79(18): 4577-4579, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31519775

RESUMO

Understanding the way hypoxia influences tumor biology is important; to study tumor hypoxia, simple and robust quantification of tissue oxygenation levels in vivo is necessary. Real-time noninvasive imaging without the use of expensive large equipment (PET or MRI) is most desirable. Photoacoustic imaging, in the form of volumetric multispectral optoacoustic tomography, as described in this issue of Cancer Research, paves the way for tumor hypoxia studies using an intrinsic optical contrast agent (hemoglobin), up to cm depth and 0.1-mm spatial resolution, in real-time 3D. This approach may find use in the clinic to assess tumor status and therapeutic efficacy.See related article by Ron et al., p. 4767.


Assuntos
Neoplasias , Técnicas Fotoacústicas , Meios de Contraste , Humanos , Hipóxia , Hipóxia Tumoral
9.
Nat Commun ; 10(1): 4277, 2019 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-31537800

RESUMO

Chronic cranial window (CCW) is an essential tool in enabling longitudinal imaging and manipulation of various brain activities in live animals. However, an active CCW capable of sensing the concealed in vivo environment while simultaneously providing longitudinal optical access to the brain is not currently available. Here we report a disposable ultrasound-sensing CCW (usCCW) featuring an integrated transparent nanophotonic ultrasonic detector fabricated using soft nanoimprint lithography process. We optimize the sensor design and the associated fabrication process to significantly improve detection sensitivity and reliability, which are critical for the intend longitudinal in vivo investigations. Surgically implanting the usCCW on the skull creates a self-contained environment, maintaining optical access while eliminating the need for external ultrasound coupling medium for photoacoustic imaging. Using this usCCW, we demonstrate photoacoustic microscopy of cortical vascular network in live mice over 28 days. This work establishes the foundation for integrating photoacoustic imaging with modern brain research.


Assuntos
Encéfalo/diagnóstico por imagem , Técnicas Fotoacústicas/métodos , Crânio/cirurgia , Ultrassonografia/métodos , Animais , Camundongos , Camundongos Endogâmicos C57BL
10.
Opt Express ; 27(14): 19369-19381, 2019 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-31503697

RESUMO

Photoacoustic endomicroscopy (PAEM) is capable of imaging fine structures in digestive tract. However, conventional PAEM employs a tightly focused laser beam to irradiate the object, which results in a limited depth-of-field (DOF). Here, we propose a scanning-domain synthesis of optical beams (SDSOB) to optimize both transverse resolution and the DOF by synthetic effective focused beams in scanning domain for the PAEM. By utilizing the SDSOB technique, multiple defocused and scattered beams are refocused to synthesize virtual focuses covering a large range of depth. A transverse point spread function that is 5.7-time sharper, and a transverse spatial bandwidth that is 8.5-time broader than those of the conventional PAEM were simulatively obtained through SDSOB-PAEM at the defocus distance of 2.4 mm. We validated the transverse resolution improvement at both in- and out-focus positions via phantom experiments of carbon fibers. In addition, in vivo rabbit experiments were conducted to acquire vascular images over radial depth range of 900 µm. And further morphological analysis revealed that the SDSOB images were acquired with abundant vascular branches and nodes, large total-length and small average-length of blood vessels, which indicated that the SDSOB-PAEM achieved high-resolution imaging in distinct rectal layers. All these results suggest that the SDSOB-PAEM possesses high transverse resolution and extended DOF, which demonstrates the SDSOB-PAEM can provide more accurate information for clinical assessment.


Assuntos
Microscopia , Fenômenos Ópticos , Técnicas Fotoacústicas/métodos , Animais , Simulação por Computador , Feminino , Análise Numérica Assistida por Computador , Imagens de Fantasmas , Coelhos
11.
J Clin Neurosci ; 70: 242-246, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31477467

RESUMO

Accurate margin delineation and safe maximal resection of glioma is one of the most challenging problems of neurosurgery, due to its close resemblance to normal brain parenchyma. However, different intraoperative visualization methods have been used for real-time intraoperative investigation of the borders of the resection cavity, each having advantages and limitations. This preliminary study was designed to simulate multi-wavelength photoacoustic imaging for brain tumor margin delineation for maximum safe resection of glioma. Since the photoacoustic signal is directly related to the amount of optical energy absorption by the endogenous tissue chromophores such as hemoglobin; it may be able to illustrate the critical structures such as tumor vessels during surgery. The simulation of the optical and acoustic part was done by using Monte-Carlo and k-wave toolbox, respectively. As our simulation results proved, at different wavelengths and depths, the amount of optical absorption for the blood layer is significantly different from others such as normal and tumoral tissues. Furthermore, experimental validation of our approach confirms that, by using multi-wavelengths proportional to the depth of the tumor margin during surgery, tumor margin can be differented using photoacoustic imaging at various depths. Photoacoustic imaging may be considered as a promising imaging modality which combines the spectral contrast of optical imaging as well as the spatial resolution of ultrasound imaging, and may be able to delineate the vascular-rich glioma margins at different depths of the resection cavity during surgery.


Assuntos
Neoplasias Encefálicas/diagnóstico por imagem , Glioma/diagnóstico por imagem , Neuroimagem/métodos , Técnicas Fotoacústicas/métodos , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/cirurgia , Glioma/patologia , Glioma/cirurgia , Humanos , Margens de Excisão , Método de Monte Carlo
12.
Opt Lett ; 44(16): 4063-4066, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31415547

RESUMO

Photo-mediated ultrasound therapy (PUT) is a novel, noninvasive antimicrovascular approach that can treat neovascularization with high precision. We developed a photoacoustic (PA) sensing (PAS) system for PUT and achieved real-time PAS-guided PUT. Experiments performed on a chicken yolk sac membrane model demonstrated that PAS could monitor the treatment effect in a microvessel during PUT. Vessel shrinkage induced a decrease in the PA signal amplitude, while vessel rupture induced an abrupt increase in the PA signal amplitude. The integrated PUT and PAS system can significantly improve the safety and effectiveness of PUT, and may assist with clinical translation of this novel antimicrovascular technique.


Assuntos
Técnicas Fotoacústicas , Terapia por Ultrassom/métodos , Fenômenos Ópticos , Razão Sinal-Ruído , Fatores de Tempo
14.
Chem Commun (Camb) ; 55(73): 10940-10943, 2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31441920

RESUMO

Different from traditional "always on" photothermal therapy (PTT) agents, tumor microenvironment responsive agents showed more tumor specificity and lower photo-toxicity to normal tissues. Herein, a photo-stable and reversible pH responsive phenazine dye (PIOH) was synthesized and assembled with liposomes forming nanoparticles (PIOH-NPs), which exhibited a strong NIR absorption in a weak acid environment and were successfully utilized for photoacoustic (PA) imaging-guided photothermal therapy in mice.


Assuntos
Antineoplásicos/uso terapêutico , Corantes Fluorescentes/uso terapêutico , Fenazinas/uso terapêutico , Animais , Antineoplásicos/síntese química , Antineoplásicos/química , Antineoplásicos/toxicidade , Colesterol/química , Feminino , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/química , Corantes Fluorescentes/toxicidade , Concentração de Íons de Hidrogênio , Hipertermia Induzida/métodos , Lecitinas/química , Lipossomos/química , Camundongos Endogâmicos BALB C , Nanopartículas/química , Tamanho da Partícula , Fenazinas/síntese química , Fenazinas/química , Fenazinas/toxicidade , Técnicas Fotoacústicas/métodos , Fototerapia/métodos , Microambiente Tumoral/fisiologia
15.
Nanoscale ; 11(34): 16080-16091, 2019 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-31432846

RESUMO

An injectable multifunctional hydrogel based on an engineered coiled-coil polypeptide, Ag2S quantum dots (QDs), and paclitaxel (PTX) has been developed for sustained chemo-photothermal therapy. Oil-soluble Ag2S QDs and PTX were first loaded into nanogels formed with engineered polypeptide PC10A by ultrasonic treatment to prepare PC10A/Ag2S QD/PTX nanogels. The multifunctional PC10A/Ag2S QD/PTX hydrogels were prepared by dissolving the PC10A/Ag2S QD/PTX nanogels into the PC10A hydrogel. The PC10A/Ag2S QD/PTX hydrogel can be injected directly into the site of tumors. In vitro and in vivo toxicity results showed that the PC10A/Ag2S QD/PTX hydrogel presented excellent biocompatibility. Compared with single near-infrared photothermal therapy and chemotherapy, the combined therapy could effectively suppress the growth of SKOV3 ovarian carcinoma tumor. In addition, real-time monitoring of the in vivo degradation of the PC10A/Ag2S QD/PTX hydrogel was achieved by near-infrared fluorescence imaging and photoacoustic imaging. These results demonstrated that this injectable multifunctional PC10A/Ag2S QD/PTX hydrogel has the potential as a theranostic platform for sustained cancer treatments.


Assuntos
Hidrogéis , Hipertermia Induzida , Neoplasias Experimentais , Imagem Óptica , Neoplasias Ovarianas , Técnicas Fotoacústicas , Fototerapia , Pontos Quânticos , Animais , Linhagem Celular Tumoral , Feminino , Humanos , Hidrogéis/química , Hidrogéis/farmacologia , Camundongos , Camundongos Nus , Neoplasias Experimentais/diagnóstico por imagem , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/terapia , Neoplasias Ovarianas/diagnóstico por imagem , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/terapia , Pontos Quânticos/química , Pontos Quânticos/uso terapêutico
16.
Opt Lett ; 44(17): 4119-4122, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31465343

RESUMO

Detailed assessment of skin conditions or the efficacy of skin treatments could greatly benefit from noninvasively assessing the distribution of cutaneous and subcutaneous structures and biomolecules. We considered ultrawideband raster scan optoacoustic mesoscopy with an extended wavelength range from visible to short-wave infrared and observed previously unseen high-resolution images of lipids colocalized with water, melanin, and hemoglobin distribution in human skin. Based on this contrast, the technique resolves subcutaneous fat, the pilosebaceous unit with complete hair strand and bulb, dermal microvasculature, and epidermal structures. We further visualize melanoidins that form via the Maillard reaction in the ultrathin stratum corneum layer, analyze their absorption spectrum, and separate them from the melanin layer. The suggested method may allow novel interrogation of skin conditions, possibly impacting diagnostics and medical and cosmetic treatments.


Assuntos
Raios Infravermelhos , Fenômenos Ópticos , Técnicas Fotoacústicas , Pele/diagnóstico por imagem , Tecido Adiposo/citologia , Hemoglobinas/metabolismo , Humanos , Metabolismo dos Lipídeos , Melaninas/metabolismo , Pele/citologia , Pele/metabolismo , Água/metabolismo
17.
J Biomed Nanotechnol ; 15(10): 2090-2099, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31462373

RESUMO

Chemodynamic therapy based on Fenton or Fenton-like reactions is an emerging cancer therapy that has attracted considerable attention due to its high selectivity for tumors. However, the low catalytic efficiency of the Fenton or Fenton-like agent limits its therapeutic effect. Combination therapies based on chemodynamic therapy using "all-in-one" agents show great potential for cancer treatment. In this study, W18O49 nanorods with strong near-infrared absorption and excellent Fenton-like reaction performance were prepared, and their application in photoacoustic imaging-guided photothermal-chemodynamic combination cancer therapy was explored. First, hydrophobic W18O49 nanorods were synthesized via a one-step pyrolysis method. Hydrophilic W18O49 nanorods were then prepared via hydrophobic self-assembly between oleyl amine-coated W18O49 nanorods and an amphiphilic polymer. The W18O49 nanorods exhibited good photoacoustic, photothermal, and chemodynamic performance in vitro along with good photoacoustic imaging contrast and excellent therapeutic effect in photothermal-chemodynamic combination cancer therapy in vivo. This study provides a low-cost and easy-to-prepare agent for photothermal-chemodynamic combination therapy and promotes the application of photothermal-chemodynamic combination therapy in cancer treatment.


Assuntos
Nanotubos , Neoplasias , Técnicas Fotoacústicas , Terapia Combinada , Humanos , Polímeros
18.
Opt Lett ; 44(15): 3773-3776, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31368965

RESUMO

Hypoxia, a low tissue oxygenation condition caused by insufficient oxygen supply, leads to potentially irreversible tissue damage, such as brain infarction during stroke. Intravascular oxygenation has long been used by photoacoustic imaging, among other imaging modalities, to study hypoxia. However, intravascular oxygenation describes only the oxygen supply via microcirculation, which does not directly reflect the amount of free oxygen available for metabolism in the interstitial fluid. Therefore, to fully understand hypoxia, it is highly desirable to monitor blood oxygenation as well as tissue oxygenation during the same biological process. In this work, by combining high-resolution photoacoustic microscopy (PAM) and a novel bioreducible N-oxide-based hypoxia-sensitive probe HyP-650, we have demonstrated simultaneous imaging of intravascular oxygenation and tissue hypoxia. We have established detailed chemical, optical, and photoacoustic properties of HyP-650 for hypoxic activation in vitro and in living cells. We have also performed PAM on hindlimb ischemia models and tumor-bearing mice to study the correlation between intravascular oxygenation and tissue oxygenation at various hypoxic levels. We expect that Hyp-650 enhanced photoacoustic imaging will find a variety of applications in brain and cancer research.


Assuntos
Vasos Sanguíneos/diagnóstico por imagem , Vasos Sanguíneos/metabolismo , Oxigênio/metabolismo , Técnicas Fotoacústicas/métodos , Animais , Membro Posterior/irrigação sanguínea , Isquemia/metabolismo , Isquemia/patologia , Camundongos , Microscopia , Hipóxia Tumoral
19.
Opt Lett ; 44(15): 3897-3900, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31368996

RESUMO

We present photoacoustic shadow-casting microscopy (PASM), a technique that allows high-resolution imaging of weakly absorbing biological samples with unprecedented sensitivity. In PASM, a uniform optical absorbing layer is placed in contact with the samples and is excited by the light transmitted through the sample, producing photoacoustic (PA) waves with an increased signal-to-noise ratio compared with that generated by the sample itself. Therefore, given a desired image quality, the required excitation fluence is much reduced, alleviating the photothermal damage to the specimen. The system provides a lateral resolution of 5 µm when using a 0.30 NA microscope objective lens. To demonstrate PASM, we present images of bovine red blood cells and microbeads.


Assuntos
Microscopia/métodos , Técnicas Fotoacústicas/métodos , Animais , Bovinos , Eritrócitos/citologia , Razão Sinal-Ruído
20.
Vox Sang ; 114(7): 701-710, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31392743

RESUMO

BACKGROUND AND OBJECTIVES: During the in vitro storage of red blood cells (RBCs), unfavourable changes (storage lesions) cause a rapid consumption of intracellular diphosphoglycerate. The latter deregulates the oxygen-haemoglobin binding potential, subsequently increasing oxygen saturation (SO2 ) and membrane degradation, transforming RBCs from biconcave discs to rigid spherical bodies (spheroechinocytes). Current laboratory techniques invasively extract RBC samples to assess the quality of red cell concentrate (RCC) units. Optical technologies could provide a means of assessing quality non-invasively. MATERIALS AND METHODS: A photoacoustic (PA) imaging technique was developed for acquiring the SO2 of blood bags non-invasively. Seven RCC units were monitored every 3-5 days until expiry (6 weeks). Measurements were validated against a conventional blood gas analyzer (BGA). Using an image flow cytometry assay, morphological profile trends were compared against the SO2 trends during blood bag storage. RESULTS: A strong correlation (r2  ≥ 0·95) was found when comparing temporal data between PA and BGA SO2 measurements. Inter-sample PA variability was found to be similar to that produced by BGA (±0·8%). A strong correlation was found to exist between the temporal changes in SO2 and relative spheroechinocyte population (0·79 ≤ r2  ≤ 0·97). CONCLUSION: This study suggests that PA imaging can non-invasively track the SO2 of stored RBCs non-invasively. By longitudinally monitoring the change in SO2 , it is possible to infer the effects of the storage lesion on RBC morphology. This non-invasive monitoring technique allows for the assessment of blood bags, without compromising sterility pre-transfusion.


Assuntos
Preservação de Sangue/normas , Técnicas Fotoacústicas/métodos , Preservação de Sangue/métodos , Eritrócitos/citologia , Estudos de Viabilidade , Citometria de Fluxo/métodos , Humanos
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