وأيد هذا العمل في جزء من مؤسسة العلوم الوطنية (CBET 0846453).

<ol>
	<li>McDonald, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prototyping+of+microfluidic+devices+in+poly(dimethylsiloxane)+using+solid-object+printing.">Prototyping of microfluidic devices in poly(dimethylsiloxane) using solid-object printing.</a> Anal Chem. 74, 1537-1545 (2002).</li><li>Hansen, M. C., Palmer, R. J. J., Udsen, C., White, D. C., Molin, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+GFP+fluorescence+in+cells+of+Streptococcus+gordonii+under+conditions+of+low+pH+and+low+oxygen+concentration.">Assessment of GFP fluorescence in cells of Streptococcus gordonii under conditions of low pH and low oxygen concentration.</a> Microbiol. 147, 1383-1391 (2001).</li><li>Englert, D. L., Manson, M. D., Jayaraman, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Flow-Based+Microfluidic+Device+for+Quantifying+Bacterial+Chemotaxis+in+Stable,+Competing+Gradients.">A Flow-Based Microfluidic Device for Quantifying Bacterial Chemotaxis in Stable, Competing Gradients.</a> Appl Environ Microbiol. 75, 4557-4564 (2009).</li><li>Englert, D. L., Jayaraman, A., Manson, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Microfluidic techniques for the analysis of bacterial chemotaxis. , (2009).</li><li>Mao, H., Cremer, P. S., &amp;amp, M. a. n. s. o. n. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+sensitive,+versatile+microfluidic+assay+for+bacterial+chemotaxis.">A sensitive, versatile microfluidic assay for bacterial chemotaxis.</a> PNAS. 100 (9), 5449-5454 (2003).</li></ol>

ويمكن حساب معامل القسم الكيميائي (CPC) ومعامل الهجرة الكيميائي (CMC) كما هو موضح في ماو وآخرون 5. إذا تم الكشف عن خلية على الجانب عالية التركيز ، وإعطائها قيمة +1 ، في حين يرد الكشف عن خلية على الجانب تركيز منخفض قيمة -1. ولخص القيم صعودا ومقسوما على العدد الكلي للخلايا لتوليد الحزب الشيوعى الصينى. علامة للحزب الشيوعى الصينى (إيجابية أو سلبية) ويعطي اتجاه الهجرة (نحو أو بعيدا عن إشارة). على الرغم من ان الحزب الشيوعى الصينى يشير إلى ما إذا كانت الخلايا تستجيب لمادة كيميائية باعتبارها جاذبة أو طاردة ، فإنه لا يفعل قياس مدى انجذاب كيميائي. لهذا ، فإننا حساب اللجنة العسكرية المركزية من خلال تقسيم ملف التوزيع المكاني من البكتيريا عبر عرض الجهاز إلى 64 أقسام (32 على كل من جانبي مدخل الخلية) ، وتعيين عامل الترجيح إلى الخلايا في كل مقطع على أساس مسافة الهجرة. تتضاعف المقاطع ابعد من المركز (المادتين 1 و 64) بواسطة +1 (= 31.5/31.5) أو -1 لأنها تحتوي على الخلايا التي هاجروا المسافة القصوى. تتضاعف المقبل قسمين (2 و 63) بواسطة +0.968 (= 30.5/31.5) أو -0.968. تتضاعف القسمين الأخيرين (أي 32 و 33 التي هي الأقرب إلى مدخل الخلية) من 0.015 + (= 0.5/31.5) أو -0.015. ولخص القيم المرجحة للتطبيع والعدد الكلي للخلايا لتوليد CMC. A +1 CMC من يشير إلى أن جميع هذه البكتيريا انتقلت تماما عن الانحدار إلى جدار الغرفة التدفق. في حالة وجود استجابة جذابة خفيفة ، واللجنة العسكرية المركزية لا تزال إيجابية ، ولكن لها قيمة أصغر ، في حين أن القيمة للحزب الشيوعى الصينى سيظل +1. وبالتالي CMC يميز الخلايا تستجيب على أساس حد للهجرة. معايير معينة يجب أن يتم التحكم بدقة أثناء أداء التجارب الكيميائي. درجة الحرارة التي تزرع البكتيريا هو المهم. لهاء القولونية ، لاحظنا أن درجة حرارة أفضل نمو هي 32 درجة مئوية والحد من ارتفاع درجات الحرارة الحركية. لمستقبلات معينة لتكون موجودة في البكتيريا ، قد يكون من الضروري لنمو البكتيريا في وجود حمل chemoeffector (أي رئيس للخلايا الكيميائي). على سبيل المثال ، عند التحقيق في مدى استجابة البكتيريا لالمالتوز ، تزرع الخلايا بنسبة 0.1 ٪ (وزن / حجم) من السكر. عند إعادة التعليق على البكتيريا بعد الطرد المركزي ، فمن المهم أن يتم تنفيذ رقيقة تهز / المتداول من الأنبوب وليست vortexed الخلايا. هز قوي يمكن القص وسياط وتقليص القدرة على الحركة من الخلايا التي يجري اختبارها. فإن معدلات تدفق المستخدمة في الجهاز يجب أن يتم تحديد استنادا إلى الحركية للبكتيريا التي يجري اختبارها. قد تكون هناك حاجة لمعدلات أبطأ تدفق البكتيريا التي هي أقل متحركة ، ويجب أن تحدد معدل التدفق الأمثل تجريبيا. ونحن نتوقع أن يمكن استخدام الجهاز لإجراء تحقيقات μFlow الأساسية على الجرثومي الكيميائي (على سبيل المثال ، المنافسة بين chemoeffectors لمستقبلات نفسه) وكذلك في تحديد البكتيريا في العينات البيئية التي تستجيب للمواد الكيميائية خاصة فيما جاذبة أو طاردة.

a microfluidic device for quantifying bacterial chemotaxis in stable concentration gradients

الكيميائي يسمح البكتيريا لنهج مصادر جاذبة للمواد الكيميائية أو لتجنب مصادر للمواد الكيميائية طارد. رصد البكتيريا باستمرار تركيز chemoeffectors محددة بمقارنة التركز الحالي للتركيز الكشف عن بضع ثوان في وقت سابق. هذه المقارنة يحدد اتجاه صافي الحركة. على الرغم من عدة تدرجات المتنافسة غالبا ما تتعايش في الطبيعة ، والنهج التقليدي للتحقيق الكيميائي الجرثومي غير كافية لقياس الهجرة ردا على التدرجات تركيز الجاذبة والطاردة. هنا ، نحن تصف وضع نموذج لتقديم الكيميائي ميكروفلويديك التدرجات تركيز دقيق وثابت من chemoeffectors للبكتيريا والتحقيق كميا استجابتها لتطبيق التدرج. الجهاز تنوعا في تلك التدرجات تركيز تركيز أي المطلقة المطلوبة وقوة الانحدار يمكن توليدها بسهولة عن طريق خلط ناشر. وأظهر الجهاز باستخدام استجابة<em&gt; القولونية</em&gt; RP437 إلى التدرجات من الأحماض الأمينية وأيونات النيكل.

Watch this Scientific Journal Video about A Microfluidic Device for Quantifying Bacterial Chemotaxis in Stable Concentration Gradients at JoVE.com

A Microfluidic Device for Quantifying Bacterial Chemotaxis in Stable Concentration Gradients

يصف هذا البروتوكول على تطوير جهاز للتحقيق في ميكروفلويديك الكيميائي الجرثومي في التدرجات تركيز chemoeffectors مستقرة.

جهاز ميكروفلويديك الانجذاب الكيميائي لقياس تركيز البكتيريا في التدرجات مستقرة

Chemotaxis allows bacteria to approach sources of attractant chemicals or to avoid sources of repellent chemicals. Bacteria constantly monitor the ...

a-microfluidic-device-for-quantifying-bacterial-chemotaxis-stable

Nanyang Technological University

Research

JoVE Journal

Biology

12.1K Views.  Texas A&M University. يصف هذا البروتوكول على تطوير جهاز للتحقيق في ميكروفلويديك الكيميائي الجرثومي في التدرجات تركيز chemoeffectors مستقرة.

Video: A Microfluidic Device for Quantifying Bacterial Chemotaxis in Stable Concentration Gradients

Chemotactic Response of Marine Micro-Organisms to Micro-Scale Nutrient Layers

The degree to which planktonic microbes can exploit microscale resource patches will have considerable implications for oceanic trophodynamics and biogeochemical flux. However, to take advantage of nutrient patches in the ocean, swimming microbes must overcome the influences of physical forces including molecular diffusion and turbulent shear, which will limit the availability of patches and the ability of bacteria to locate them. Until recently, methodological limitations have precluded direct examinations of microbial behaviour within patchy habitats and realistic small-scale flow conditions. Hence, much of our current knowledge regarding microbial behaviour in the ocean has been procured from theoretical predictions. To obtain new information on microbial foraging behaviour in the ocean we have applied soft lithographic fabrication techniques to develop 2 microfluidic devices, which we have used to create (i) microscale nutrient patches with dimensions and diffusive characteristics relevant to oceanic processes and (ii) microscale vortices, with shear rates corresponding to those expected in the ocean. These microfluidic devices have permitted a first direct examination of microbial swimming and chemotactic behaviour within a heterogeneous and dynamic seascape. The combined use of epifluorescence and phase contrast microscopy allow direct examinations of the physical dimensions and diffusive characteristics of nutrient patches, while observing the population-level aggregative response, in addition to the swimming behaviour of individual microbes. These experiments have revealed that some species of phytoplankton, heterotrophic bacteria and phagotrophic protists are adept at locating and exploiting diffusing microscale resource patches within very short time frames. We have also shown that up to moderate shear rates, marine bacteria are able to  fight the flow  and swim through their environment at their own accord. However, beyond a threshold high shear level, bacteria are aligned in the shear flow and are less capable of swimming without disturbance from the flow. Microfluidics represents a novel and inexpensive approach for studying aquatic microbial ecology, and due to its suitability for accurately creating realistic flow fields and substrate gradients at the microscale, is ideally applicable to examinations of microbial behaviour at the smallest scales of interaction.  We therefore suggest that microfluidics represents a valuable tool for obtaining a better understanding of the ecology of microorganisms in the ocean.

The fabrication of microfluidic channels and their implementation in experiments for studying the chemotactic foraging behaviour of marine microbes within a patchy nutrient seascape and the swimming behaviour of bacteria within shear flow are described.

الكيميائي استجابة الكائنات المجهرية البحرية الصغيرة الحجم لطبقات غذائية

The degree to which planktonic microbes can exploit microscale resource patches will have considerable implications for oceanic trophodynamics and ...

Studies of Bacterial Chemotaxis Using Microfluidics - Interview

دراسات لاستخدام الانجذاب الكيميائي الجرثومي على microfluidics -- مقابلة

A Gradient-generating Microfluidic Device for Cell Biology

 The fabrication and operation of a gradient-generating microfluidic device for studying cellular behavior is described.  A microfluidic platform is an enabling experimental tool, because it can precisely manipulate fluid flows, enable high-throughput experiments, and generate stable soluble concentration gradients.  Compared to conventional gradient generators, poly(dimethylsiloxane) (PDMS)-based  microfluidic devices can generate stable concentration gradients of growth factors with well-defined profiles.  Here, we developed simple gradient-generating microfluidic devices with three separate inlets.  Three microchannels combined into one microchannel to generate concentration gradients.  The stability and shape of growth factor gradients were confirmed by fluorescein isothyiocyanate (FITC)-dextran with a molecular weight similar to epidermal growth factor (EGF).  Using this microfluidic device, we demonstrated that fibroblasts exposed to concentration gradients of EGF migrated toward higher concentrations.  The directional orientation of cell migration and motility of migrating cells were quantitatively assessed by cell tracking analysis.  Thus, this gradient-generating microfluidic device might be useful for studying and analyzing the behavior of migrating cells.

We describe a protocol for the microfabrication of the gradient-generating microfluidic device that can generate spatial and temporal gradients in well-defined microenvironment. In this approach, the gradient-generating microfluidic device can be used to study directed cell migration, embryogenesis, wound healing, and cancer metastasis.

جهاز لتوليد التدرج ميكروفلويديك لبيولوجيا الخلية

 The fabrication and operation of a gradient-generating microfluidic device for studying cellular behavior is described.  A microfluidic platform is an ...

Microfluidic Platform for Measuring Neutrophil Chemotaxis from Unprocessed Whole Blood

Neutrophils play an essential role in protection against infections and their numbers in the blood are frequently measured in the clinic. Higher neutrophil counts in the blood are usually an indicator of ongoing infections, while low neutrophil counts are a warning sign for higher risks for infections. To accomplish their functions, neutrophils also have to be able to move effectively from the blood where they spend most of their life, into tissues, where infections occur. Consequently, any defects in the ability of neutrophils to migrate can increase the risks for infections, even when neutrophils are present in appropriate numbers in the blood. However, measuring neutrophil migration ability in the clinic is a challenging task, which is time consuming, requires large volume of blood, and expert knowledge. To address these limitations, we designed a robust microfluidic assays for neutrophil migration, which requires a single droplet of unprocessed blood, circumvents the need for neutrophil separation, and is easy to quantify on a simple microscope. In this assay, neutrophils migrate directly from the blood droplet, through small channels, towards the source of chemoattractant. To prevent the granular flow of red blood cells through the same channels, we implemented mechanical filters with right angle turns that selectively block the advance of red blood cells. We validated the assay by comparing neutrophil migration from blood droplets collected from finger prick and venous blood. We also compared these whole blood (WB) sources with neutrophil migration from samples of purified neutrophils and found consistent speed and directionality between the three sources. This microfluidic platform will enable the study of human neutrophil migration in the clinic and the research setting to help advance our understanding of neutrophil functions in health and disease.

This protocol details an assay designed to measure human neutrophil chemotaxis from one droplet of whole blood with robust reproducibility. This approach circumvents the need for neutrophil separation and requires only a few minutes of assay preparation time. The microfluidic chip enables the repeated measure of neutrophil chemotaxis over time in infants or small mammals, where sample volume is limited.

منصة ميكروفلويديك الانجذاب الكيميائي لقياس العدلات من الدم غير المجهزة الجامعة

Neutrophils play an essential role in protection against infections and their numbers in the blood are frequently measured in the clinic. Higher ...

Bioengineering

In Situ Chemotaxis Assay to Examine Microbial Behavior in Aquatic Ecosystems

Microbial behaviors, such as motility and chemotaxis (the ability of a cell to alter its movement in response to a chemical gradient), are widespread across the bacterial and archaeal domains. Chemotaxis can result in substantial resource acquisition advantages in heterogeneous environments. It also plays a crucial role in symbiotic interactions, disease, and global processes, such as biogeochemical cycling. However, current techniques restrict chemotaxis research to the laboratory and are not easily applicable in the field. Presented here is a step-by-step protocol for the deployment of the in situ chemotaxis assay (ISCA), a device that enables robust interrogation of microbial chemotaxis directly in the natural environment. The ISCA is a microfluidic device consisting of a 20 well array, in which chemicals of interest can be loaded. Once deployed in aqueous environments, chemicals diffuse out of the wells, creating concentration gradients that microbes sense and respond to by swimming into the wells via chemotaxis. The well contents can then be sampled and used to (1) quantify strength of the chemotactic responses to specific compounds through flow cytometry, (2) isolate and culture responsive microorganisms, and (3) characterize the identity and genomic potential of the responding populations through molecular techniques. The ISCA is a flexible platform that can be deployed in any system with an aqueous phase, including marine, freshwater, and soil environments.

Presented here is the protocol for an in situ chemotaxis assay, a recently developed microfluidic device that enables studies of microbial behavior directly in the environment.

في الموقع Chemotaxis اختبار لفحص السلوك الميكروبي في النظم الإيكولوجية المائية

Microbial behaviors, such as motility and chemotaxis (the ability of a cell to alter its movement in response to a chemical gradient), are widespread ...

Environment

An All-on-chip Method for Rapid Neutrophil Chemotaxis Analysis Directly from a Drop of Blood

Neutrophil migration and chemotaxis are critical for our body's immune system. Microfluidic devices are increasingly used for investigating neutrophil migration and chemotaxis owing to their advantages in real-time visualization, precise control of chemical concentration gradient generation, and reduced reagent and sample consumption. Recently, a growing effort has been made by the microfluidic researchers toward developing integrated and easily operated microfluidic chemotaxis analysis systems, directly from whole blood. In this direction, the first all-on-chip method was developed for integrating the magnetic negative purification of neutrophils and the chemotaxis assay from small blood volume samples. This new method permits a rapid sample-to-result neutrophil chemotaxis test in 25 min. In this paper, we provide detailed construction, operation and data analysis method for this all-on-chip chemotaxis assay with a discussion on troubleshooting strategies, limitations and future directions. Representative results of the neutrophil chemotaxis assay testing a defined chemoattractant, N-Formyl-Met-Leu-Phe (fMLP), and sputum from a chronic obstructive pulmonary disease (COPD) patient, using this all-on-chip method are shown. This method is applicable to many cell migration-related investigations and clinical applications.

This article provides the detailed method of performing a rapid neutrophil chemotaxis assay by integrating the on-chip neutrophil isolation from whole blood and the chemotaxis test on a single microfluidic chip.

طريقة الكل على رقاقة للتحليل الكيميائي العدلات السريعة مباشرة من قطرة الدم

Neutrophil migration and chemotaxis are critical for our body's immune system. Microfluidic devices are increasingly used for investigating neutrophil ...

Immunology and Infection

Traction Microscopy Integrated with Microfluidics for Chemotactic Collective Migration

Cells change migration patterns in response to chemical stimuli, including the gradients of the stimuli. Cellular migration in the direction of a chemical gradient, known as chemotaxis, plays an important role in development, the immune response, wound healing, and cancer metastasis. While chemotaxis modulates the migration of single cells as well as collections of cells in vivo, in vitro research focuses on single-cell chemotaxis, partly due to the lack of the proper experimental tools. To fill that gap, described here is a unique experimental system that combines microfluidics and micropatterning to demonstrate the effects of chemical gradients on collective cell migration. Furthermore, traction microscopy and monolayer stress microscopy are incorporated into the system to characterize changes in cellular force on the substrate as well as between neighboring cells. As proof-of-concept, the migration of micropatterned circular islands of Madin-Darby canine kidney (MDCK) cells is tested under a gradient of hepatocyte growth factor (HGF), a known scattering factor. It is found that cells located near the higher concentration of HGF migrate faster than those on the opposite side within a cell island. Within the same island, cellular traction is similar on both sides, but intercellular stress is much lower on the side of higher HGF concentration. This novel experimental system can provide new opportunities to studying the mechanics of chemotactic migration by cellular collectives.

Collective cell migration in development, wound healing, and cancer metastasis is often guided by the gradients of growth factors or signaling molecules. Described here is an experimental system combining traction microscopy with a microfluidic system and a demonstration of how to quantify the mechanics of collective migration under biochemical gradient.

المجهر الجر المتكاملة مع ميكروفلويديكس للهجرة الجماعية الكيميائية

Cells change migration patterns in response to chemical stimuli, including the gradients of the stimuli. Cellular migration in the direction of a chemical ...

Generating Controlled, Dynamic Chemical Landscapes to Study Microbial Behavior

We demonstrate a method for the generation of controlled, dynamic chemical pulses&#8213;where localized chemoattractant becomes suddenly available at the microscale&#8213;to create micro-environments for microbial chemotaxis experiments. To create chemical pulses, we developed a system to introduce amino acid sources near-instantaneously by photolysis of caged amino acids within a polydimethylsiloxane (PDMS) microfluidic chamber containing a bacterial suspension. We applied this method to the chemotactic bacterium, Vibrio ordalii, which can actively climb these dynamic chemical gradients while being tracked by video microscopy. Amino acids, rendered biologically inert (&#39;caged&#39;) by chemical modification with a photoremovable protecting group, are uniformly present in the suspension but not available for consumption until their sudden release, which occurs at user-defined points in time and space by means of a near-UV-A focused LED beam. The number of molecules released in the pulse can be determined by a calibration relationship between exposure time and uncaging fraction, where the absorption spectrum after photolysis is characterized by using UV-Vis spectroscopy. A nanoporous polycarbonate (PCTE) membrane can be integrated into the microfluidic device to allow the continuous removal by flow of the uncaged compounds and the spent media. A strong, irreversible bond between the PCTE membrane and the PDMS microfluidic structure is achieved by coating the membrane with a solution of 3-aminopropyltriethoxysilane (APTES) followed by plasma activation of the surfaces to be bonded. A computer-controlled system can generate user-defined sequences of pulses at different locations and with different intensities, so as to create resource landscapes with prescribed spatial and temporal variability. In each chemical landscape, the dynamics of bacterial movement at the individual scale and their accumulation at the population level can be obtained, thereby allowing the quantification of chemotactic performance and its effects on bacterial aggregations in ecologically relevant environments.

A protocol for the generation of dynamic chemical landscapes by photolysis within microfluidic and millifluidic setups is presented. This methodology is suitable to study diverse biological processes, including the motile behavior, nutrient uptake, or adaptation to chemicals of microorganisms, both at the single cell and population level.

توليد المناظر الطبيعية الكيميائية الديناميكية الخاضعة للرقابة لدراسة السلوك الميكروبي

We demonstrate a method for the generation of controlled, dynamic chemical pulses&#8213;where localized chemoattractant becomes suddenly available at the ...

Measurement of Cellular Chemotaxis with ECIS/Taxis

Cellular movement in response to external stimuli is fundamental to many cellular processes including wound healing, inflammation and the response to infection. A common method to measure chemotaxis is the Boyden chamber assay, in which cells and chemoattractant are separated by a porous membrane. As cells migrate through the membrane toward the chemoattractant, they adhere to the underside of the membrane, or fall into the underlying media, and are subsequently stained and visually counted 1. In this method, cells are exposed to a steep and transient chemoattractant gradient, which is thought to be a poor representation of gradients found in tissues 2.
Another assay system, the under-agarose chemotaxis assay, 3, 4 measures cell movement across a solid substrate in a thin aqueous film that forms under the agarose layer. The gradient that develops in the agarose is shallow and is thought to be an appropriate representation of naturally occurring gradients. Chemotaxis can be evaluated by microscopic imaging of the distance traveled. Both the Boyden chamber assay and the under-agarose assay are usually configured as endpoint assays.
The automated ECIS/Taxis system combines the under-agarose approach with Electric Cell-substrate Impedance Sensing (ECIS) 5, 6. In this assay, target electrodes are located in each of 8 chambers. A large counter-electrode runs through each of the 8 chambers (Figure 2). Each chamber is filled with agarose and two small wells are the cut in the agarose on either side of the target electrode. One well is filled with the test cell population, while the other holds the sources of diffusing chemoattractant (Figure 3). Current passed through the system can be used to determine the change in resistance that occurs as cells pass over the target electrode. Cells on the target electrode increase the resistance of the system 6. In addition, rapid fluctuations in the resistance represent changes in the interactions of cells with the electrode surface and are indicative of ongoing cellular shape changes. The ECIS/Taxis system can measure movement of the cell population in real-time over extended periods of time, but is also sensitive enough to detect the arrival of a single cell at the target electrode. 
Dictyostelium discoidium is known to migrate in the presence of a folate gradient 7, 8 and its chemotactic response can be accurately measured by ECIS/Taxis 9. Leukocyte chemotaxis, in response to SDF1&alpha; and to chemotaxis antagonists has also been measured with ECIS/Taxis 10, 11. An example of the leukocyte response to SDF1&alpha; is shown in Figure 1.

The ECIS/Taxis system is an automated, real-time assay that measures cellular chemotaxis. In this assay, cells move beneath a layer of agarose to arrive at a target electrode. Cellular movement is measured by the onset of resistance to AC current 0.

قياس من الانجذاب الكيميائي الخلوي مع ECIS / سيارات الأجرة

Cellular movement in response to external stimuli is fundamental to many cellular processes including wound healing, inflammation and the response to ...

Imaging G Protein-coupled Receptor-mediated Chemotaxis and its Signaling Events in Neutrophil-like HL60 Cells

Eukaryotic cells sense and move towards a chemoattractant gradient, a cellular process referred as chemotaxis. Chemotaxis plays critical roles in many physiological processes, such as embryogenesis, neuron patterning, metastasis of cancer cells, recruitment of neutrophils to sites of inflammation, and the development of the model organism Dictyostelium discoideum. Eukaryotic cells sense chemo-attractants using G protein-coupled receptors. Visual chemotaxis assays are essential for a better understanding of how eukaryotic cells control chemoattractant-mediated directional cell migration. Here, we describe detailed methods for: 1) real-time, high-resolution monitoring of multiple chemotaxis assays, and 2) simultaneously visualizing the chemoattractant gradient and the spatiotemporal dynamics of signaling events in neutrophil-like HL60 cells.

Visual chemotaxis assays are essential for a better understanding of how eukaryotic cells control chemoattractant-mediated directional cell migration. Here, we describe detailed methods for: 1) real-time, high-resolution monitoring of multiple chemotaxis assays, and 2) simultaneously visualizing the chemoattractant gradient and the spatiotemporal dynamics of signaling events in neutrophil-like HL60 cells.

يقترن تصوير البروتين G مستقبلات بوساطة الكيميائي والأحداث يشير في خلايا HL60 العدلات مثل

Eukaryotic cells sense and move towards a chemoattractant gradient, a cellular process referred as chemotaxis. Chemotaxis plays critical roles in many ...