وأيد هذا العمل من قبل المعاهد الوطنية للصحة منح (R01 HL086699 ، HL086699 - 01A2S1 ، 1S10RR027327 - 01) إلى MM. معتمد جزئيا مقالتنا التي Microimaging كارل زايس م.

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الإعلان عن أي تضارب في المصالح.

وصف طريقة القياس هنا يسمح السريع وكمية من أنواع الاكسجين التفاعلية في الخلايا الحية إما باستخدام الأصباغ الأكسدة حساسة أو أجهزة الاستشعار البلازميد. منبهات من TLRs (مثل مستقبلات تول) هي المركبات التي تحفز الخلايا من خلال TLRs موجودة على سطح الخلية وتحريك مسارات إشارات ا...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> الكاشف </td><td> شركة </td><td> فهرس العدد </td></tr><tr><td> Attofluor الخلية الغرفة </td><td> Invitrogen </td><td> A7816 </td></tr><tr><td> Antimycin A </td><td> سيغما الدريتش </td><td> A8674 </td></tr><tr><td> DMEM المتوسطة انخفاض الجلوكوز </td><td> Invitrogen </td><td> 10567-014 </td></tr><tr><td> عامل النمو البطاني تكملة (رسم القلب) </td><td> ريف </td><td> 02-102 </td></tr><tr><td> المصل البقري الجنين </td><td> Invitrogen </td><td> 12662011 </td></tr><tr><td> G418 </td><td> Invitrogen </td><td> 10131-027 </td></tr><tr><td> الجيلاتين </td><td> سيغما الدريتش </td><td> G1393 </td></tr><tr><td> H 2 DCFDA </td><td> Invitrogen </td><td> D - 399 </td></tr><tr><td> LPS </td><td> سيغما الدريتش </td><td> L4516 </td></tr><tr><td> LTA </td><td> سيغما الدريتش </td><td> L2515 </td></tr><tr><td> MitoSOX الأحمر </td><td> Invitrogen </td><td> M36008 </td></tr><tr><td> OPTI - MEM أنا انخفاض متوسط ​​المصل </td><td> Invitrogen </td><td> 51985091 </td></tr><tr><td> القلم / بكتيريا (10X) </td><td> Invitrogen </td><td> 15140163 </td></tr><tr><td> pHyPer - خلوي </td><td> Evrogen </td><td> FP941 </td></tr><tr><td> pHyPer - dMito </td><td> Evrogen </td><td> FP942 </td></tr><tr><td> بولي (I : C) </td><td> سيغما الدريتش </td><td> P0913 </td></tr><tr><td> المنشور 5.0 البرمجيات </td><td> GraphPad البرمجيات ، وشركة </td><td></td></tr><tr><td> SigmaPlot 11.0 </td><td> Systat البرمجيات ، المؤتمر الوطني العراقي. </td><td></td></tr><tr><td> التربسين - EDTA (10X) </td><td> Invitrogen </td><td> 15400054 </td></tr><tr><td> T - 25 قوارير </td><td> كورنينج </td><td> 430639 </td></tr><tr><td> T - 75 قوارير </td><td> فالكون دينار بحريني </td><td> 353136 </td></tr><tr><td> 96 - T جيداC الصغرى لوحة جيدا </td><td> فالكون دينار بحريني </td><td> 353072 </td></tr><tr><td> زن 2009 البرمجيات </td><td> كارل زايس المجهري ميتا 510 مبائر </td><td></td></tr></tbody></table>

visualization of vascular ca2+ signaling triggered by paracrine derived ros

وقد تورط الاكسدة في عدد من الظروف المرضية بما في ذلك نقص التروية / ضخه الأضرار والتسمم. مفهوم الاكسدة يشير إلى تشكيل الشاذة من ريوس (أنواع الاكسجين التفاعلية) ، والتي تشمل • O 2 -- ، H 2 O 2 ، وجذور الهيدروكسيل. أنواع الاكسجين التفاعلية التأثيرات العديد من العمليات الخلوية بما في ذلك نقل الإشارة ، وانتشار الخلايا وموت الخلايا 1-6. ROS لديها القدرة على الأوعية الدموية وتلف خلايا الجهاز مباشرة ، ويمكن الشروع في التفاعلات الكيميائية الثانوية والتعديلات الوراثية التي تؤدي في نهاية المطاف في التضخيم من الأضرار الأولية بوساطة نسيج ريوس. ومن المكونات الرئيسية لتتالي التضخيم الذي لا رجعة فيه تفاقم تلف الأنسجة هو توظيف وتفعيل تعميم الخلايا الالتهابية. أثناء الالتهاب ، وإنتاج السيتوكينات الالتهابية الخلايا مثل نخر الورم عامل α (TNFα) ، وأنه IL - 1تنشيط الخلايا البطانية (EC) والخلايا الظهارية ، وكذلك زيادة 7 الاستجابة الالتهابية. خلل بطانة الأوعية الدموية هي ميزة أنشئت من التهاب حاد. الضامة تساهم في اختلال وظيفي التهاب بطانة خلال الآليات التي لا تزال غامضة. تفعيل نتائج الضامة خارج الخلايا في الإفراج عن O 2 • -- والعديد من السيتوكينات الموالية للالتهابات ، والتي يطلق إشارات مرضية في الخلايا المجاورة 8. oxidases NADPH هي المصدر الرئيسي والأساسي للروس في معظم أنواع الخلايا. في الآونة الأخيرة ، فإنه يظهر من قبلنا وغيرها 9،10 ريوس أن تنتجها oxidases NADPH تحفز إنتاج ريوس الميتوكوندريا خلال ظروف الفيزيولوجية المرضية كثيرة. وبالتالي قياس إنتاج ريوس الميتوكوندريا هي نفس القدر من الأهمية ، بالإضافة إلى قياس عصاري خلوي ريوس. الضامة تنتج ريوس من NADPH أوكسيديز انزيم بروتين فلافيني الذي يلعب دورا رئيسيا في التهاب. تنشيط مرة واحدة ،NADPH أوكسيديز أكلة تنتج كميات وفيرة من O 2 • -- التي تعتبر مهمة في الآلية الدفاعية للمضيف 11،12. على الرغم من نظير الصماوي المستمدة من O 2 • -- يلعب دورا هاما في التسبب في أمراض الأوعية الدموية ، والتصور من الإشارات التي يسببها ريوس نظير الصماوي الخلايا بما في ذلك تعبئة كا 2 + لا تزال الفرضية. وقد طورنا نموذجا والتي تستخدم في تنشيط البلاعم كمصدر لل• O 2 -- لتنبيغ إشارة إلى الخلايا البطانية المجاورة. باستخدام هذا النموذج الذي يثبت أن بلعم المستمدة من O 2 • -- أن يؤدي إلى إشارات الكالسيوم في الخلايا البطانية المجاورة.

Watch this Scientific Journal Video about Visualization of Vascular Ca2+ Signaling Triggered by Paracrine Derived ROS at JoVE.com

Visualization of Vascular Ca2+ Signaling Triggered by Paracrine Derived ROS

وصفت وسيلة فعالة للحصول على نظرة ثاقبة الرؤية والاستقراء ريوس نظير الصماوي المستمدة من البطانية إشارة + CA2. هذا الأسلوب يستفيد من قياس نظير الصماوي ريوس المستمدة أثار CA2 + تعبئة الخلايا البطانية في الأوعية الدموية في نموذج شارك في الثقافة.

تصور كا الأوعية الدموية 2 + اشارة سببها ريوس المشتقة نظير الصماوي

Oxidative stress has been implicated in a number of pathologic conditions including ischemia/reperfusion damage and sepsis. The concept of oxidative ...

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Visualization of Vascular Ca2+ Signaling Triggered by Paracrine Derived ROS

15.4K Views.  Temple University. وصفت وسيلة فعالة للحصول على نظرة ثاقبة الرؤية والاستقراء ريوس نظير الصماوي المستمدة من البطانية إشارة + CA2. هذا الأسلوب يستفيد من قياس نظير الصماوي ريوس المستمدة أثار CA2 + تعبئة الخلايا البطانية في الأوعية الدموية في نموذج شارك في الثقافة.

Video: Visualization of Vascular Ca2+ Signaling Triggered by Paracrine Derived ROS

Techniques for Imaging Ca2+ Signaling in Human Sperm

Fluorescence microscopy of cells loaded with fluorescent, Ca2+-sensitive dyes is used for measurement of spatial and temporal aspects of Ca2+ signaling in live cells. Here we describe the method used in our laboratories for loading suspensions of human sperm with Ca2+-reporting dyes and measuring the fluorescence signal during physiological stimulation. Motile cells are isolated by direct swim-up and incubated under capacitating conditions for 0-24 h, depending upon the experiment. The cell-permeant AM (acetoxy methyl ester) ester form of the Ca2+-reporting dye is then added to a cell aliquot and a period of 1 h is allowed for loading of the dye into the cytoplasm. We use visible wavelength dyes to minimize photo-damage to the cells, but this means that ratiometric recording is not possible. Advantages and disadvantages of this approach are discussed. During the loading period cells are introduced into an imaging chamber and allowed to adhere to a poly-D-lysine coated coverslip. At the end of the loading period excess dye and loose cells are removed by connection of the chamber to the perfusion apparatus. The chamber is perfused continuously, stimuli and modified salines are then added to the perfusion header. Experiments are recorded by time-lapse acquisition of fluorescence images and analyzed in detail offline, by manually drawing regions of interest. Data are normalized to pre-stimulus levels such that, for each cell (or part of a cell), a graph showing the Ca2+ response as % change in fluorescence is obtained.

Techniques for Imaging Ca2+ Signaling in Human Sperm

Stimulus-evoked [Ca2+]i signals of individual human sperm are assessed. Motile cells are loaded with Ca2+-sensitive fluorescent dye (AM-ester method) and immobilised in a perfusable chamber. Cells are imaged by time-lapse fluorescence microscopy and stimulated via the perfusing medium. Responses of single cells (or regions) are analysed offline using Excel.

تقنيات التصوير كاليفورنيا 2 + في اشارة الى الحيوانات المنوية البشرية

Fluorescence microscopy of cells loaded with fluorescent, Ca2+-sensitive dyes is used for measurement of spatial and temporal aspects of Ca2+ signaling in ...

Visualization of ATP Synthase Dimers in Mitochondria by Electron Cryo-tomography

Electron cryo-tomography is a powerful tool in structural biology, capable of visualizing the three-dimensional structure of biological samples, such as cells, organelles, membrane vesicles, or viruses at molecular detail. To achieve this, the aqueous sample is rapidly vitrified in liquid ethane, which preserves it in a close-to-native, frozen-hydrated state. In the electron microscope, tilt series are recorded at liquid nitrogen temperature, from which 3D tomograms are reconstructed. The signal-to-noise ratio of the tomographic volume is inherently low. Recognizable, recurring features are enhanced by subtomogram averaging, by which individual subvolumes are cut out, aligned and averaged to reduce noise. In this way, 3D maps with a resolution of 2 nm or better can be obtained. A fit of available high-resolution structures to the 3D volume then produces atomic models of protein complexes in their native environment. Here we show how we use electron cryo-tomography to study the in situ organization of large membrane protein complexes in mitochondria. We find that ATP synthases are organized in rows of dimers along highly curved apices of the inner membrane cristae, whereas complex I is randomly distributed in the membrane regions on either side of the rows. By subtomogram averaging we obtained a structure of the mitochondrial ATP synthase dimer within the cristae membrane.

We present a protocol of how to collect and process electron cryo-tomograms of whole mitochondria. The technique provides detailed insights into the structure, function, and organization of large membrane protein complexes in native biological membranes.

التصور من ATP سينسيز Dimers في الميتوكوندريا بواسطة الكترون البرد المقطعي

Electron cryo-tomography is a powerful tool in structural biology, capable of visualizing the three-dimensional structure of biological samples, such as ...

Fecal Glucocorticoid Analysis: Non-invasive Adrenal Monitoring in Equids

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, corticotrophin releasing hormone (CRH) is released from the hypothalamus in the brain. This stimulates the release of adrenocorticotrophic hormone (ACTH) from the pituitary gland, which in turn stimulates release of glucocorticoids from the adrenal gland. In horses the glucocorticoid corticosterone is responsible for several adaptations needed to support equine flight behaviour and subsequent removal from the aversive situation. Corticosterone metabolites can be detected in the feces of horses and assessment offers a non-invasive option to evaluate long term patterns of adrenal activity. Fecal assessment offers advantages over other techniques that monitor adrenal activity including blood plasma and saliva analysis. The non-invasive nature of the method avoids sampling stress which can confound results. It also allows the opportunity for repeated sampling over time and is ideal for studies in free ranging horses. This protocol describes the enzyme linked immunoassay (EIA) used to assess feces for corticosterone, in addition to the associated biochemical validation.

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. The method offers a non-invasive option to assess long term patterns in both domestic and free ranging horses. This protocol describes the enzyme linked immunoassay involved and the associated biochemical validation.

برازي تحليل جلايكورتيكود: رصد الغدة الكظرية غير الغازية في الخيليات

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, ...

DiI Perfusion as a Method for Vascular Visualization in Ambystoma mexicanum

Perfusion techniques have been used for centuries to visualize the circulation of tissues. Axolotl (Ambystoma mexicanum) is a species of salamander that has emerged as an essential model for regeneration studies. Little is known about how revascularization occurs in the context of regeneration in these animals. Here we report a simple method for visualization of the vasculature in axolotl via perfusion of 1,1&#8217;-Dioctadecy-3,3,3&#8217;,3&#8217;-tetramethylindocarbocyanine perchlorate (DiI). DiI is a lipophilic carbocyanine dye that inserts into the plasma membrane of endothelial cells instantaneously. Perfusion is done using a peristaltic pump such that DiI enters the circulation through the aorta. During perfusion, dye flows through the axolotl&#8217;s blood vessels and incorporates into the lipid bilayer of vascular endothelial cells upon contact. The perfusion procedure takes approximately one hour for an eight-inch axolotl. Immediately after perfusion with DiI, the axolotl can be visualized with a confocal fluorescent microscope. The DiI emits light in the red-orange range when excited with a green fluorescent filter. This DiI perfusion procedure can be used to visualize the vascular structure of axolotls or to demonstrate patterns of revascularization in regenerating tissues.

DiI Perfusion as a Method for Vascular Visualization in Ambystoma mexicanum

Using a lipophilic 1,1&#39;-Dioctadecy-3,3,3&#39;,3&#39;-tetramethylindocarbocyanine perchlorate (DiI) staining technique, Ambystoma mexicanum can undergo vascular perfusion to allow for easy visualization of the vasculature.

الجاذبة الإرواء كطريقة لتصور الأوعية الدموية في<em&gt; أمبيستوما ميكسيكانوم</em

Perfusion techniques have been used for centuries to visualize the circulation of tissues. Axolotl (Ambystoma mexicanum) is a species of salamander that ...

Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography

This protocol describes how to visualize the transient DNA compaction in cyanobacteria. DNA compaction is a dramatic cytoplasmic event recently found to occur in some cyanobacteria before cell division. However, due to the large cell size and the transient character, it is difficult to investigate the structure in detail. To overcome the difficulties, first, DNA compaction is reproducibly produced in the cyanobacterium Synechococcus elongatus PCC 7942 by synchronous culture using 12 h each light/dark cycle. Second, DNA compaction is monitored by fluorescence microscopy and captured by rapid freezing. Third, the detailed structure of DNA compacted cells is visualized in three dimensions (3D) by high voltage cryo-electron tomography. This set of methods is widely applicable to investigate transient structures in bacteria, e.g. cell division, chromosome segregation, phage infection etc., which are monitored by fluorescence microscopy and directly visualized by cryo-electron tomography at appropriate time points.

This protocol describes how to visualize the transient DNA compaction in cyanobacteria. Synchronous cultivation, monitoring by fluorescence microscopy, rapid freezing, and high voltage cryo-electron tomography are used. A protocol for these methodologies is presented, and future applications and developments are discussed.

التصور للضغط الحمض النووي في البكتيريا الزرقاء بالتصوير المقطعي الفولت العالي Cryo-إلكترون

This protocol describes how to visualize the transient DNA compaction in cyanobacteria. DNA compaction is a dramatic cytoplasmic event recently found to ...

Molecular Analysis of Endothelial-mesenchymal Transition Induced by Transforming Growth Factor-&#946; Signaling

Phenotypic plasticity of endothelial cells underlies cardiovascular system development, cardiovascular diseases, and various conditions associated with organ fibrosis. In these conditions, differentiated endothelial cells acquire mesenchymal-like phenotypes. This process is called endothelial-mesenchymal transition (EndMT) and is characterized by downregulation of endothelial markers, upregulation of mesenchymal markers, and morphological changes. EndMT is induced by several signaling pathways, including transforming growth factor (TGF)-&#946;, Wnt, and Notch, and regulated by molecular mechanisms similar to those of epithelial-mesenchymal transition (EMT) important for gastrulation, tissue fibrosis, and cancer metastasis. Understanding the mechanisms of EndMT is important to develop diagnostic and therapeutic approaches targeting EndMT. Robust induction of EndMT in vitro is useful to characterize common gene expression signatures, identify druggable molecular mechanisms, and screen for modulators of EndMT. Here, we describe an in vitro method for induction of EndMT. MS-1 mouse pancreatic microvascular endothelial cells undergo EndMT after prolonged exposure to TGF-&#946; and show upregulation of mesenchymal markers and morphological changes as well as induction of multiple inflammatory chemokines and cytokines. Methods for the analysis of microRNA (miRNA) modulation are also included. These methods provide a platform to investigate mechanisms underlying EndMT and the contribution of miRNAs to EndMT.

Molecular Analysis of Endothelial-mesenchymal Transition Induced by Transforming Growth Factor-&amp;#946; Signaling

A protocol for in vitro induction of endothelial-mesenchymal transition (EndMT), which is useful for investigating cellular signaling pathways involved in EndMT, is described. In this experimental model, EndMT is induced by treatment with TGF-&#946; in MS-1 endothelial cells.

التحليل الجزيئي غشائي الوسيطة الانتقالية الناجمة عن تحويل الإشارات عامل النمو-β

Phenotypic plasticity of endothelial cells underlies cardiovascular system development, cardiovascular diseases, and various conditions associated with ...

Retinal Cryo-sections, Whole-Mounts, and Hypotonic Isolated Vasculature Preparations for Immunohistochemical Visualization of Microvascular Pericytes

Retinal pericytes play an important role in many diseases of the eye. Immunohistochemical staining techniques of retinal vessels and microvascular pericytes are central to ophthalmological research. It is vital to choose an appropriate method of visualizing the microvascular pericytes. We describe retinal microvascular pericyte immunohistochemical staining in cryo-sections, whole-mounts, and hypotonic isolated vasculature using antibodies for platelet-derived growth factor receptor &#946; (PDGFR&#946;) and nerve/glial antigen 2 (NG2). This allows us to highlight advantages and shortcomings of each of the three tissue preparations for the visualization of the retinal microvascular pericytes. Cryo-sections provide transsectional visualization of all retinal layers but contain only a few occasional transverse cuts of the microvasculature. Whole-mount provides an overview of the entire retinal vasculature, but visualization of the microvasculature can be troublesome. Hypotonic isolation provides a method to visualize the entire retinal vasculature by the removal of neuronal cells, but this makes the tissue very fragile.

We demonstrate three different tissue preparation techniques for immunohistochemical visualization of rat retinal microvascular pericytes, i.e., cryo-sections, whole-mounts, and hypotonic isolation of the vascular network.

البرد الشبكية-الفروع، الجامع--يتصاعد، والتحضيرات ناقص التوتر المفرج معزولة للتصور المناعي من بيريسيتيس ميكروفاسكولار

Retinal pericytes play an important role in many diseases of the eye. Immunohistochemical staining techniques of retinal vessels and microvascular ...

A Proximal Culture Method to Study Paracrine Signaling Between Cells

Intercellular interactions play an important role in many biological processes, including tumor progression, immune responses, angiogenesis, and development. Paracrine or juxtacrine signaling mediates such interactions. The use of a conditioned medium and coculture studies are the most common methods to discriminate between these two types of interactions. However, the effect of localized high concentrations of secreted factors in the microenvironment during the paracrine interactions is not accurately recapitulated by conditioned medium and, thus, may lead to imprecise conclusions. To overcome this problem, we have devised a proximal culture method to study paracrine signaling. The two cell types are grown on either surface of a 10 &#181;m-thick polycarbonate membrane with 0.4 &#181;m pores. The pores allow the exchange of secreted factors and, at the same time, inhibit juxtacrine signaling. The cells can be collected and lysed at the endpoint to determine the effects of the paracrine signaling. In addition to allowing for localized concentration gradients of secreted factors, this method is amenable to experiments involving prolonged periods of culture, as well as the use of inhibitors. While we use this method to study the interactions between ovarian cancer cells and the mesothelial cells they encounter at the site of metastasis, it can be adapted to any two adherent cell types for researchers to study paracrine signaling in various fields, including tumor microenvironment, immunology, and development.

Paracrine and juxtacrine cellular interactions play an important role in many biological processes, including tumor progression, immune responses, angiogenesis, and development. Here, a proximal culture method is used to study paracrine signaling where the localized concentrations of the secreted factors are maintained while preventing direct cellular contact.

أسلوب ثقافة الدانية لدراسة Paracrine الإشارات بين الخلايا

Intercellular interactions play an important role in many biological processes, including tumor progression, immune responses, angiogenesis, and ...

Myeloid Innate Signaling Pathway Regulation by MALT1 Paracaspase Activity

Besides its function in lymphoid cells, which has been addressed by numerous studies, the paracaspase MALT1 also plays an important role in innate cells downstream of pattern recognition receptors. Best studied are the Dectin-1 and Dectin-2 members of the C-type lectin-like receptor family that induce a SYK- and CARD9-dependent signaling cascade leading to NF-&#954;B activation, in a MALT1-dependent manner. By contrast, Toll-like receptors (TLR), such as TLR-4, propagate NF-&#954;B activation but signal via an MYD88/IRAK-dependent cascade. Nonetheless, whether MALT1 might contribute to TLR-4 signaling has remained unclear. Recent evidence with MLT-827, a potent and selective inhibitor of MALT1 paracaspase activity, indicates that TNF- production downstream of TLR-4 in human myeloid cells is independent of MALT1, as opposed to TNF- production downstream of Dectin-1, which is MALT1 dependent. Here, we addressed the selective involvement of MALT1 in pattern recognition sensing further, using a variety of human and mouse cellular preparations, and stimulation of Dectin-1, MINCLE or TLR-4 pathways. We also provided additional insights by exploring cytokines beyond TNF-, and by comparing MLT-827 to a SYK inhibitor (Cpd11) and to an IKK inhibitor (AFN700). Collectively, the data provided further evidence for the MALT1-dependency of C-type lectin-like receptor &#8212;signaling by contrast to TLR-signaling.

MALT1 regulates innate immunity but how this occurs remains ill-defined. We used the selective MALT1 paracaspase inhibitor MLT-827 to unravel the contribution of MALT1 to innate signaling downstream of Toll-like or C-type lectin-like receptors, demonstrating that MALT1 regulates the production of myeloid cytokines, and downstream of C-type lectin-like receptors, selectively.

تنظيم مسار الإشارات الفطرية النقوي بنشاط باراكاسباسي MALT1

Besides its function in lymphoid cells, which has been addressed by numerous studies, the paracaspase MALT1 also plays an important role in innate cells ...

Applications of Spatio-temporal Mapping and Particle Analysis Techniques to Quantify Intracellular Ca2+ Signaling In Situ

Ca2+ imaging of isolated cells or specific types of cells within intact tissues often reveals complex patterns of Ca2+ signaling. This activity requires careful and in-depth analyses and quantification to capture as much information about the underlying events as possible. Spatial, temporal and intensity parameters intrinsic to Ca2+ signals such as frequency, duration, propagation, velocity and amplitude may provide some biological information required for intracellular signalling. High-resolution Ca2+ imaging typically results in the acquisition of large data files that are time consuming to process in terms of translating the imaging information into quantifiable data, and this process can be susceptible to human error and bias. Analysis of Ca2+ signals from cells in situ typically relies on simple intensity measurements from arbitrarily selected regions of interest (ROI) within a field of view (FOV). This approach ignores much of the important signaling information contained in the FOV. Thus, in order to maximize recovery of information from such high-resolution recordings obtained with Ca2+dyes or optogenetic Ca2+ imaging, appropriate spatial and temporal analysis of the Ca2+ signals is required. The protocols outlined in this paper will describe how a high volume of data can be obtained from Ca2+ imaging recordings to facilitate more complete analysis and quantification of Ca2+ signals recorded from cells using a combination of spatiotemporal map (STM)-based analysis and particle-based analysis. The protocols also describe how different patterns of Ca2+ signaling observed in different cell populations in situ can be analyzed appropriately. For illustration, the method will examine Ca2+ signaling in a specialized population of cells in the small intestine, interstitial cells of Cajal (ICC), using GECIs.

Applications of Spatio-temporal Mapping and Particle Analysis Techniques to Quantify Intracellular Ca2+ Signaling In Situ

Genetically encoded Ca2+ indicators (GECIs) have radically changed how in situ Ca2+ imaging is performed. To maximize data recovery from such recordings, appropriate analysis of Ca2+ signals is required. The protocols in this paper facilitate the quantification of Ca2+ signals recorded in situ using spatiotemporal mapping and particle-based analysis.

تطبيقات رسم الخرائط الزمانية وتقنيات تحليل الجسيمات للتحديد الكمي لداخل الخلية Ca2 + إشارات في الموقع

Ca2+ imaging of isolated cells or specific types of cells within intact tissues often reveals complex patterns of Ca2+ signaling. This activity requires ...