IntraVital最新文献_第2页

A novel model for ectopic, chronic, intravital multiphoton imaging of bone marrow vasculature and architecture in split femurs. 一个新的模型异位，慢性，活体多光子成像骨髓血管和结构的分裂股骨。

IntraVital Pub Date : 2015-06-30 eCollection Date: 2015-05-01 DOI: 10.1080/21659087.2015.1066949

Mirela Bălan, Friedemann Kiefer

{"title":"A novel model for ectopic, chronic, intravital multiphoton imaging of bone marrow vasculature and architecture in split femurs.","authors":"Mirela Bălan, Friedemann Kiefer","doi":"10.1080/21659087.2015.1066949","DOIUrl":"https://doi.org/10.1080/21659087.2015.1066949","url":null,"abstract":"Creating a model for intravital visualization of femoral bone marrow, a major site of hematopoiesis in adult mammalian organisms, poses a serious challenge, in that it needs to overcome bone opacity and the inaccessibility of marrow. Furthermore, meaningful analysis of bone marrow developmental and differentiation processes requires the repetitive observation of the same site over long periods of time, which we refer to as chronic imaging. To surmount these issues, we developed a chronic intravital imaging model that allows the observation of split femurs, ectopically transplanted into a dorsal skinfold chamber of a host mouse. Repeated, long term observations are facilitated by multiphoton microscopy, an imaging technique that combines superior imaging capacity at greater tissue depth with low phototoxicity. The transplanted, ectopic femur was stabilized by its sterile environment and rapidly connected to the host vasculature, allowing further development and observation of extended processes. After optimizing transplant age and grafting procedure, we observed the development of new woven bone and maturation of secondary ossification centers in the transplanted femurs, preceded by the sprouting of a sinusoidal-like vascular network, which was almost entirely composed of femoral endothelial cells. After two weeks, the transplant was still populated with stromal and haematopoietic cells belonging both to donor and host. Over this time frame, the transplant partially retained myeloid progenitor cells with single and multi-lineage differentiation capacity. In summary, our model allowed repeated intravital imaging of bone marrow angiogenesis and hematopoiesis. It represents a promising starting point for the development of improved chronic optical imaging models for femoral bone marrow.","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"4 2","pages":"e1066949"},"PeriodicalIF":0.0,"publicationDate":"2015-06-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21659087.2015.1066949","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Imaging CD8⁺ T cells during diverse viral infections. 在多种病毒感染过程中对 CD8+ T 细胞进行成像。

IntraVital Pub Date : 2015-06-19 eCollection Date: 2015-01-01 DOI: 10.1080/21659087.2015.1055425

Heather D Hickman

{"title":"Imaging CD8+ T cells during diverse viral infections.","authors":"Heather D Hickman","doi":"10.1080/21659087.2015.1055425","DOIUrl":"10.1080/21659087.2015.1055425","url":null,"abstract":"CD8+ T cells play a critical role in host defense against pathogens and tumors. Much of our current knowledge of the activation and subsequent effector activities of CD8+ T cells has been gained using ex vivo approaches examining the T cell population en masse for surface phenotype, activation status and the production of effector molecules. Thus, the precise behaviors and diversity of individual CD8+ T cells responding to virus infection in vivo have not been extensively explored, leaving many unanswered questions relevant to the rational design of antiviral vaccines and therapeutics. Recently, intravital multiphoton microscopy (MPM) has been used to image CD8+ T cell priming after infection with disparate viral pathogens ranging from small RNA viruses encoding few proteins to DNA viruses producing hundreds of viral proteins (many immunomodulatory). After priming, effector CD8+ T cells have been visualized in virus-infected tissue, both during primary infection and after transitioning to tissue resident memory cells (TRM). Here, I highlight recent advances in our understanding of antiviral CD8+ T cell responses revealed through intravital MPM.","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"4 1","pages":"e1055425"},"PeriodicalIF":0.0,"publicationDate":"2015-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5226004/pdf/kinv-04-01-1055425.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769369","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

In vivo imaging of the tumor and its associated microenvironment using combined CARS / 2-photon microscopy. 利用 CARS 和双光子显微镜对肿瘤及其相关微环境进行活体成像。

IntraVital Pub Date : 2015-06-08 eCollection Date: 2015-01-01 DOI: 10.1080/21659087.2015.1055430

Martin Lee, Andy Downes, You-Ying Chau, Bryan Serrels, Nick Hastie, Alistair Elfick, Valerie Brunton, Margaret Frame, Alan Serrels

引用次数: 0

The caspase 3 sensor Phiphilux G2D2 is activated non-specifically in S1 renal proximal tubules caspase 3传感器philphilux G2D2在S1肾近端小管中非特异性激活

IntraVital Pub Date : 2015-05-04 DOI: 10.1080/21659087.2015.1067352

T. Hato, R. Sandoval, P. Dagher

引用次数: 3

The Use of Second Harmonic Generation to Image the Extracellular Matrix During Tumor Progression. 肿瘤进展过程中利用二次谐波成像细胞外基质。

IntraVital Pub Date : 2015-01-06 eCollection Date: 2014-12-01 DOI: 10.4161/21659087.2014.984509

Kathleen Burke, Edward Brown

引用次数: 20

Intravital imaging of dendritic spine plasticity. 树突脊柱可塑性的活体成像。

IntraVital Pub Date : 2015-01-06 eCollection Date: 2014-12-01 DOI: 10.4161/21659087.2014.984504

Cora Sau Wan Lai

引用次数: 2

Flying back to the nest: Intravital microscopy reveals how the niche can induce stemness. 飞回巢穴：肉眼显微镜揭示了生态位如何诱导干性。

IntraVital Pub Date : 2014-08-11 eCollection Date: 2014-01-01 DOI: 10.4161/intv.29653

Narges M Rashidi, Cristina Lo Celso

引用次数: 0

Imaging windows for long-term intravital imaging: General overview and technical insights. 长期活体成像的成像窗口:一般概述和技术见解。

IntraVital Pub Date : 2014-08-11 eCollection Date: 2014-01-01 DOI: 10.4161/intv.29917

Maria Alieva, Laila Ritsma, Randy J Giedt, Ralph Weissleder, Jacco van Rheenen

{"title":"Imaging windows for long-term intravital imaging: General overview and technical insights.","authors":"Maria Alieva, Laila Ritsma, Randy J Giedt, Ralph Weissleder, Jacco van Rheenen","doi":"10.4161/intv.29917","DOIUrl":"https://doi.org/10.4161/intv.29917","url":null,"abstract":"Intravital microscopy is increasingly used to visualize and quantitate dynamic biological processes at the (sub)cellular level in live animals. By visualizing tissues through imaging windows, individual cells (e.g., cancer, host, or stem cells) can be tracked and studied over a time-span of days to months. Several imaging windows have been developed to access tissues including the brain, superficial fascia, mammary glands, liver, kidney, pancreas, and small intestine among others. Here, we review the development of imaging windows and compare the most commonly used long-term imaging windows for cancer biology: the cranial imaging window, the dorsal skin fold chamber, the mammary imaging window, and the abdominal imaging window. Moreover, we provide technical details, considerations, and trouble-shooting tips on the surgical procedures and microscopy setups for each imaging window and explain different strategies to assure imaging of the same area over multiple imaging sessions. This review aims to be a useful resource for establishing the long-term intravital imaging procedure.","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"3 2","pages":"e29917"},"PeriodicalIF":0.0,"publicationDate":"2014-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4161/intv.29917","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769954","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 123

Quantitative intravital two-photon excitation microscopy reveals absence of pulmonary vaso-occlusion in unchallenged Sickle Cell Disease mice. 定量活体双光子激发显微镜显示无挑战镰状细胞病小鼠肺血管闭塞。

IntraVital Pub Date : 2014-07-07 DOI: 10.4161/intv.29748

Margaret F Bennewitz, Simon C Watkins, Prithu Sundd

{"title":"Quantitative intravital two-photon excitation microscopy reveals absence of pulmonary vaso-occlusion in unchallenged Sickle Cell Disease mice.","authors":"Margaret F Bennewitz, Simon C Watkins, Prithu Sundd","doi":"10.4161/intv.29748","DOIUrl":"https://doi.org/10.4161/intv.29748","url":null,"abstract":"Sickle cell disease (SCD) is a genetic disorder that leads to red blood cell (RBC) sickling, hemolysis and the upregulation of adhesion molecules on sickle RBCs. Chronic hemolysis in SCD results in a hyper-inflammatory state characterized by activation of circulating leukocytes, platelets and endothelial cells even in the absence of a crisis. A crisis in SCD is often triggered by an inflammatory stimulus and can lead to the acute chest syndrome (ACS), which is a type of lung injury and a leading cause of mortality among SCD patients. Although it is believed that pulmonary vaso-occlusion could be the phenomenon contributing to the development of ACS, the role of vaso-occlusion in ACS remains elusive. Intravital imaging of the cremaster microcirculation in SCD mice has been instrumental in establishing the role of neutrophil-RBC-endothelium interactions in systemic vaso-occlusion; however, such studies, although warranted, have never been done in the pulmonary microcirculation of SCD mice. Here, we show that two-photon excitation fluorescence microscopy can be used to perform quantitative analysis of neutrophil and RBC trafficking in the pulmonary microcirculation of SCD mice. We provide the experimental approach that enables microscopic observations under physiological conditions and use it to show that RBC and neutrophil trafficking is comparable in SCD and control mice in the absence of an inflammatory stimulus. The intravital imaging scheme proposed in this study can be useful in elucidating the cellular and molecular mechanism of pulmonary vaso-occlusion in SCD mice following an inflammatory stimulus.","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"3 2","pages":"e29748"},"PeriodicalIF":0.0,"publicationDate":"2014-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4161/intv.29748","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33322927","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 26

Lighting up microtubule cytoskeleton dynamics in skeletal muscle. 点亮骨骼肌微管细胞骨架动力学。

IntraVital Pub Date : 2014-05-30 eCollection Date: 2014-01-01 DOI: 10.4161/intv.29293

Andrius Masedunskas, Mark Appaduray, Peter W Gunning, Edna C Hardeman

{"title":"Lighting up microtubule cytoskeleton dynamics in skeletal muscle.","authors":"Andrius Masedunskas, Mark Appaduray, Peter W Gunning, Edna C Hardeman","doi":"10.4161/intv.29293","DOIUrl":"https://doi.org/10.4161/intv.29293","url":null,"abstract":"In the past few decades, live cell microscopy techniques in combination with fluorescent tagging have provided a true explosion in our knowledge of the inner functioning of the cell. Dynamic phenomena can be observed inside living cells and the behavior of individual molecules participating in those events can be documented. However, our preference for simple or easy model systems such as cell culture, has come at a cost of chasing artifacts and missing out on understanding real biology as it happens in complex multicellular organisms. We are now entering a new era where developing meaningful, but also tractable model systems to study biological phenomenon dynamically in vivo in a mammal is not only possible; it will become the gold standard for scientific quality and translational potential.1,2 A study by Oddoux et al. describing the dynamics of the microtubule (MT) cytoskeleton in skeletal muscle is one example that demonstrates the power of developing in vivo/ex vivo models.3 MTs have long attracted attention as targets for cancer therapeutics 4 and more recently as mediators of Duchene muscular dystrophy.5 The muscle fiber MT cytoskeleton forms an intricate rectilinear lattice beneath the sarcolemma and is essential for the structural integrity of the muscle. Cultured cells do not develop such a specialized organization of the MT cytoskeleton and our understanding of it has come from static snapshots of muscle sections.6 In this context, the methodology and the findings reported by Oddoux et al. are a significant step forward.","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"3 1","pages":"e29293"},"PeriodicalIF":0.0,"publicationDate":"2014-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.4161/intv.29293","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769952","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0