IntraVital最新文献

{"title":"Intravital characterization of tumor cell migration in pancreatic cancer.","authors":"Evelyne Beerling,&nbsp;Ilse Oosterom,&nbsp;Emile Voest,&nbsp;Martijn Lolkema,&nbsp;Jacco van Rheenen","doi":"10.1080/21659087.2016.1261773","DOIUrl":"https://doi.org/10.1080/21659087.2016.1261773","url":null,"abstract":"<p><p>Curing pancreatic cancer is difficult as metastases often determine the poor clinical outcome. To gain more insight into the metastatic behavior of pancreatic cancer cells, we characterized migratory cells in primary pancreatic tumors using intravital microscopy. We visualized the migratory behavior of primary tumor cells of a genetically engineered pancreatic cancer mouse model and found that pancreatic tumor cells migrate with a mesenchymal morphology as single individual cells or collectively as a stream of non-cohesive single motile cells. These findings may improve our ability to conceive treatments that block metastatic behavior.</p>","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"5 3","pages":"e1261773"},"PeriodicalIF":0.0,"publicationDate":"2016-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21659087.2016.1261773","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769370","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}

{"title":"An intravital microscopy model to study early pancreatic inflammation in type 1 diabetes in NOD mice.","authors":"Christian Lehmann,&nbsp;Nicholas B Fisher,&nbsp;Barna Tugwell,&nbsp;Juan Zhou","doi":"10.1080/21659087.2016.1215789","DOIUrl":"https://doi.org/10.1080/21659087.2016.1215789","url":null,"abstract":"<p><p>Intravital microscopy (IVM) of the pancreas has been proven to be an invaluable tool in pancreatitis, transplantation and ischemia/reperfusion research. Also in type 1 diabetes (T1D) pancreatic IVM offers unique advantages for the elucidation of the disease process. Female non-obese diabetic (NOD) mice develop T1D spontaneously by 40 weeks of age. Our goal was to establish an IVM-based method to study early pancreatic inflammation in NOD mice, which can be used to screen novel medications to prevent or delay T1D in future studies. This included evaluation of leukocyte-endothelial interactions as well as disturbances of capillary perfusion in the pancreatic microcirculation.</p>","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"5 3","pages":"e1215789"},"PeriodicalIF":0.0,"publicationDate":"2016-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21659087.2016.1215789","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769444","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}

{"title":"Direct visualization of the phenotype of hypoxic tumor cells at single cell resolution in vivo using a new hypoxia probe","authors":"Yarong Wang, Haoxuan Wang, Jiu-feng Li, D. Entenberg, Alice Xue, Weigang Wang, J. Condeelis","doi":"10.1080/21659087.2016.1187803","DOIUrl":"https://doi.org/10.1080/21659087.2016.1187803","url":null,"abstract":"ABSTRACT Tumor hypoxia is linked to tumor progression, metastasis, and therapy resistance. However, the underlying mechanisms behind this linkage are not fully understood. Here we present a novel fluorescent mCherry hypoxia-responsive marker that can be used in real time imaging to specifically and sensitively identify hypoxic cells in vivo at single cell resolution. Tumors derived from triple negative tumor cells expressing the hypoxia marker reveal that the hypoxic tumor cells congregate near flowing blood vessels. Using multiphoton microscopy, hypoxic MDA-MB-231 cells were directly visualized and showed a more persistent slow migration phenotype as compared to normoxic cells in the same field in vivo. Hypoxic tumor cells are enriched in the cell population that migrates toward human epithelial growth factor gradients in vivo, and has increased collagen degradation and intravasation activity, characteristics of dissemination and metastasis competent tumor cells. The hypoxia probe introduced in this study provides a specific reporter of hypoxic cell phenotypes in vivo which reveals new insights into the mechanisms by which hypoxia is linked to metastasis.","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"13 5","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91437582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Validation of a device for the active manipulation of the tumor microenvironment during intravital imaging","authors":"J. Williams, D. Entenberg, Yarong Wang, Álvaro Avivar-Valderas, Michael R. Padgen, Ashley Clark, J. Aguirre-Ghiso, J. Castracane, J. Condeelis","doi":"10.1080/21659087.2016.1182271","DOIUrl":"https://doi.org/10.1080/21659087.2016.1182271","url":null,"abstract":"ABSTRACT The tumor microenvironment is recognized as playing a significant role in the behavior of tumor cells and their progression to metastasis. However, tools to manipulate the tumor microenvironment directly, and image the consequences of this manipulation with single cell resolution in real time in vivo, are lacking. We describe here a method for the direct, local manipulation of microenvironmental parameters through the use of an implantable Induction Nano Intravital Device (iNANIVID) and simultaneous in vivo visualization of the results at single-cell resolution. As a proof of concept, we deliver both a sustained dose of EGF to tumor cells while intravital imaging their chemotactic response as well as locally induce hypoxia in defined microenvironments in solid tumors.","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"96 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89104524","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Intravital multiphoton imaging of mouse tibialis anterior muscle.","authors":"Jasmine Lau,&nbsp;Chi Ching Goh,&nbsp;Sapna Devi,&nbsp;Jo Keeble,&nbsp;Peter See,&nbsp;Florent Ginhoux,&nbsp;Lai Guan Ng","doi":"10.1080/21659087.2016.1156272","DOIUrl":"https://doi.org/10.1080/21659087.2016.1156272","url":null,"abstract":"<p><p>Intravital imaging by multiphoton microscopy is a powerful tool to gain invaluable insight into tissue biology and function. Here, we provide a step-by-step tissue preparation protocol for imaging the mouse tibialis anterior skeletal muscle. Additionally, we include steps for jugular vein catheterization that allow for well-controlled intravenous reagent delivery. Preparation of the tibialis anterior muscle is minimally invasive, reducing the chances of inducing damage and inflammation prior to imaging. The tibialis anterior muscle is useful for imaging leukocyte interaction with vascular endothelium, and to understand muscle contraction biology. Importantly, this model can be easily adapted to study neuromuscular diseases and myopathies.</p>","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"5 2","pages":"e1156272"},"PeriodicalIF":0.0,"publicationDate":"2016-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21659087.2016.1156272","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769443","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}

{"title":"<i>In vivo</i> imaging of skeletal muscle in mice highlights muscle defects in a model of myotubular myopathy.","authors":"Luc Mercier,&nbsp;Johann Böhm,&nbsp;Nina Fekonja,&nbsp;Guillaume Allio,&nbsp;Yves Lutz,&nbsp;Marc Koch,&nbsp;Jacky G Goetz,&nbsp;Jocelyn Laporte","doi":"10.1080/21659087.2016.1168553","DOIUrl":"https://doi.org/10.1080/21659087.2016.1168553","url":null,"abstract":"<p><p>Skeletal muscle structure and function are altered in different myopathies. However, the understanding of the molecular and cellular mechanisms mainly rely on <i>in vitro</i> and <i>ex vivo</i> investigations in mammalian models. In order to monitor <i>in vivo</i> the intracellular structure of the neuromuscular system in its environment under normal and pathological conditions, we set-up and validated non-invasive imaging of ear and leg muscles in mice. This original approach allows simultaneous imaging of different cellular and intracellular structures such as neuromuscular junctions and sarcomeres, reconstruction of the 3D architecture of the neuromuscular system, and video recording of dynamic events such as spontaneous muscle fiber contraction. Second harmonic generation was combined with vital dyes and fluorescent-coupled molecules. Skin pigmentation, although limiting, did not prevent intravital imaging. Using this versatile toolbox on the <i>Mtm1</i> knockout mouse, a model for myotubular myopathy which is a severe congenital myopathy in human, we identified several hallmarks of the disease such as defects in fiber size and neuromuscular junction shape. Intravital imaging of the neuromuscular system paves the way for the follow-up of disease progression or/and disease amelioration upon therapeutic tests. It has also the potential to reduce the number of animals needed to reach scientific conclusions.</p>","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"5 1","pages":"e1168553"},"PeriodicalIF":0.0,"publicationDate":"2016-04-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21659087.2016.1168553","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769442","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}

{"title":"Study of the cortical representation of whisker frequency selectivity using voltage-sensitive dye optical imaging.","authors":"Vassiliy Tsytsarev,&nbsp;Elena Pumbo,&nbsp;Qinggong Tang,&nbsp;Chao-Wei Chen,&nbsp;Vyacheslav Kalchenko,&nbsp;Yu Chen","doi":"10.1080/21659087.2016.1142637","DOIUrl":"https://doi.org/10.1080/21659087.2016.1142637","url":null,"abstract":"<p><p>The facial whiskers of rodents act as a high-resolution tactile apparatus that allow the animal to detect the finest details of its environment. Previously it was shown that whisker-sensitive neurons in the somatosensory cortex show frequency selectivity to small amplitude stimuli, An intravital voltage-sensitive dye optical imaging (VSDi) method in combination with the different frequency whisker stimulation was used in order to visualize neural activity in the mice somatosensory cortex in response to the stimulation of a single whisker by different frequencies. Using the intravital voltage-sensitive dye optical imaging (VSDi) method in combination with the different frequency whisker stimulation we visualized neural activity in the mice somatosensory cortex in response to the stimulation of a single whisker by different frequencies. We found that whisker stimuli with different frequencies led to different optical signals in the barrel field. Our results provide evidence that different neurons of the barrel cortex have different frequency preferences. This supports prior research that whisker deflections cause responses in cortical neurons within the barrel field according to the frequency of the stimulation. Many studies of the whisker frequency selectivity were performed using unit recording but to map spatial organization, imaging methods are essential. In the work described in the present paper, we take a serious step toward detailed functional mapping of the somatosensory cortex using VSDi. To our knowledge, this is the first demonstration of whisker frequency sensitivity and selectivity of barrel cortex neurons with optical imaging methods.</p>","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"5 1","pages":"e1142637"},"PeriodicalIF":0.0,"publicationDate":"2016-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21659087.2016.1142637","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769441","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}

{"title":"Social networking in tumor cell communities is associated with increased aggressiveness.","authors":"Catalina Lodillinsky,&nbsp;Katrina Podsypanina,&nbsp;Philippe Chavrier","doi":"10.1080/21659087.2015.1112476","DOIUrl":"https://doi.org/10.1080/21659087.2015.1112476","url":null,"abstract":"<p><p>Extracellular vesicles (EVs) are lipid-bilayer-enclosed vesicles that contain proteins, lipids and nucleic acids. EVs produced by cells from healthy tissues circulate in the blood and body fluids, and can be taken up by unrelated cells. As they have the capacity to transfer cargo proteins, lipids and nucleic acids (mostly mRNAs and miRNAs) between different cells in the body, EVs are emerging as mediators of intercellular communication that could modulate cell behavior, tissue homeostasis and regulation of physiological functions. EV-mediated cell-cell communications are also proposed to play a role in disease, for example, cancer, where they could contribute to transfer of traits required for tumor progression and metastasis. However, direct evidence for EV-mediated mRNA transfer to individual cells and for its biological consequences <i>in vivo</i> has been missing until recently. Recent studies have reported elegant experiments using genetic tracing with the Cre recombinase system and intravital imaging that visualize and quantify functional transfer of mRNA mediated by EVs in the context of cancer and metastasis.</p>","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"5 1","pages":"e1112476"},"PeriodicalIF":0.0,"publicationDate":"2016-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21659087.2015.1112476","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769438","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}

{"title":"Development of novel murine mammary imaging windows to examine wound healing effects on leukocyte trafficking in mammary tumors with intravital imaging.","authors":"Tammy Sobolik,&nbsp;Ying-Jun Su,&nbsp;Will Ashby,&nbsp;David K Schaffer,&nbsp;Sam Wells,&nbsp;John P Wikswo,&nbsp;Andries Zijlstra,&nbsp;Ann Richmond","doi":"10.1080/21659087.2015.1125562","DOIUrl":"https://doi.org/10.1080/21659087.2015.1125562","url":null,"abstract":"<p><p>We developed mammary imaging windows (MIWs) to evaluate leukocyte infiltration and cancer cell dissemination in mouse mammary tumors imaged by confocal microscopy. Previous techniques relied on surgical resection of a skin flap to image the tumor microenvironment restricting imaging time to a few hours. Utilization of mammary imaging windows offers extension of <i>intravital</i> imaging of the tumor microenvironment. We have characterized strengths and identified some previously undescribed potential weaknesses of MIW techniques. Through iterative enhancements of a transdermal portal we defined conditions for improved quality and extended confocal imaging time for imaging key cell-cell interactions in the tumor microenvironment.</p>","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"5 1","pages":"e1125562"},"PeriodicalIF":0.0,"publicationDate":"2016-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21659087.2015.1125562","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34769439","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}

{"title":"Fibroblasts keep melanoma safe from harm.","authors":"Kurt I Anderson","doi":"10.1080/21659087.2015.1074788","DOIUrl":"https://doi.org/10.1080/21659087.2015.1074788","url":null,"abstract":"The history of intravital microscopy is rooted in the study of cellular interactions in specific tissue environments. Imaging cells on glass coverslips is useful for mapping out basic features of signal transduction pathways, such as the regulation of the actin cytoskeleton by Rho-family GTPases. But the answers to many biological questions lie in the fine spatial and temporal details of signal transduction: “where” and “when” questions which can only be investigated in situ. This applies to diverse processes from T-cell / B-cell interactions within lymph nodes, to cancer-stromal cell interactions within tumors, to virtually all of embryonic development. More recently, intravital microscopy has found an important role to play in drug discovery, answering questions of where, when, and for how long drugs hit their targets at the tissue, cellular, and sub-cellular levels. This trend has been accelerated by the development of FRET biosensors which allow signal transduction to be imaged with high spatial and temporal resolution in pre-clinical cancer models. The development of B-Raf inhibitors provides an example of the promise and peril of targeted therapies, i.e. drugs designed to specifically interfere with only cancer cells. Around 50% of melanoma patients carry a mutation at V600, with the majority of these being V600E. Early clinical trials showed unprecedented improvements in overall and progression free survival of B-Raf V600E metastatic melanoma patients treated with the B-Raf inhibitor vemurafenib. These reports were accompanied by astonishing images of cancer patients riddled by metastatic melanoma being apparently cleared of their disease. Unfortunately, the benefits were short-lived and in most cases a form of melanoma returned which was completely resistant to the effects of the inhibitor. Intensive research has since uncovered several different mechanisms of acquired vemurafenib resistance, which generally involve B-Raf independent re-activation of the MAP-kinase pathway. Now, the Sahai group have used a combination of intravital microscopy and 3-dimensional culture systems to uncover a new type of drug resistance which emerges through tumor-stroma interaction. They used a FRET biosensor for ERK kinase, the terminal kinase of the MAP kinase cascade (Fig. 1) to study the response of both cancer and stromal cells to inhibition of B-Raf, the first kinase of the cascade. Surprisingly, their work shows that an off-target effect of B-Raf inhibition is activation of melanoma-associated fibroblasts (MAFs), which maintain ERK activation within the melanoma cells despite B-Raf inhibition. The MAFs do this by increasing production of extra-cellular matrix, especially fibronectin, which re-activates Erk though melanoma signaling pathways downstream of b1 integrin. The study begins with the simple observation that the B-Raf inhibitor PLX4720 impaired the growth of 2 mouse melanoma cell lines in vitro (5555 and 4434 cells), but did not retard the gro","PeriodicalId":14512,"journal":{"name":"IntraVital","volume":"4 2","pages":"e1074788"},"PeriodicalIF":0.0,"publicationDate":"2015-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21659087.2015.1074788","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34770051","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}