Bu protokol başlangıçta tasarlanmış olan laboratuar, Chicago Üniversitesi&#39;nde Dr. Aaron P. Fox teşekkür eder, ve daha da geliştirilmesi için protokol William Roden (Saint Louis Üniversitesi). Biz Whitehall Vakfı, Ulusal Bilim Vakfı ve destek için teşekkür ederiz.

Bölüm 1: Karbon Elyaf Hazırlama <ol><li> Kapiller pipet ~ 1.25 X uzunluğu bir uzunlukta bir paket karbon elyaf (7μm çapı) kesin. </li><li> Kaynama boyutlandırma kaldırın ~ 100 mL ve bir kap içinde aseton, 30 dakika veya daha uzun bir süre için kaynar aseton lifler. </li><li> Sıcak aseton lifler çıkarın ve temiz bir behere 50-100 ml taze aseton transfer. </li><li> Kuru, açık bir 150 mm Petri kabında paket yerleştirerek ve airdry geceleme fiberler. </li></ol> Bölüm 2: Yangın parlatma Cam Kapiler Tüp <ol><li> Forseps ile tüp merkezi holding ve ilgili flame.Rotate tüp sonunda açığa (uzun mikro-hematokrit kapiller tüpler, dış çapı 1.5mm, 1.3mm iç çapı 7,5 cm) Her bir kapiller tüp Yangın lehçe 3 kez alev. </li><li> Tüpü ters çevirin ve 3 kez tekrar dönen, ateşe diğer ucunu yerleştirin. </li><li> Cilalı yangın tüpleri bir cam tabak koyun ve karbon elyaf ile dolu olmak için hazır olana kadar fırın transfer. </li></ol> Bölüm 3: Kılcal Tüp içine bir Karbon Fiber Threading <ol><li> Machado ve ark açıklamalara benzer. (2008) ve Mundroff ve Wightman (2002), bir filtre ile vakum bağlı plastik boru kullanarak cam tüp içine fiber emmek. </li><li> Lif lif dışında bir tüp scalpel.Leave ¼ inç kullanarak kesin. </li></ol> Bölüm 4: Bir Elektrot Çektirme Pipet-Fiber Çekme <ol><li> Kapiler tüp elektrot çektirmenin (Narishige Japonya Model PP-830) içine yerleştirin ve yerinde kılcal tüpü tutmak için düğmeleri sıkın. </li><li> Optimal çektirmenin ayarlarında iki set çeker yürütmek için anahtarları işleyin. </li><li> Pipet yarıya çekilir sonra, çıplak fiber kesme makası kullanarak merkezinde iki yarısı ayrı. </li><li> Her elektrot yarım çıkarın ve çıkıntılı lif lif lif sadece 1 / 8 inç tüp dışında kalmaması için makas ile cam seal.Trim kırmak için çekin. </li><li> Fiber elektrotlar çekti yaklaşık 16 kadar 1-4 adımları tekrarlayın. </li></ol> Bölüm 5: Epoxy-Cam Seal yapma <ol><li> Epoksi önceden yapılmış bir kısım sıcak bir plaka ile 55 ° C sıcaklığındaki bir davlumbaz 6 dakika. </li><li> Epoksi, sargı bandı kullanarak bir paket içine çekti elektrotlar ~ 16 ısıtma iken. </li><li> EPON Reçine, 6 dakika ısıtılır sonra, 0.7 gram XOF MPDA sıcak EPON Reçine (% 14 w / v) ekleyip karıştırın flakon dönen. </li><li> Epoksi yeterli miktarda kapiller eylem yoluyla cam ipuçları girecek elektrot paket, 20 saniye ile ilgili sıcak epoksi artı sertleştirici solution.After içine batırın. </li><li> Yuvaları ile karton üzerine paket, yer elektrotlar geri alma ve bir fırın tepsiye yerleştirin 100 ° F. </li><li> En az 48 saat sonra, epoksi ile doldurmak, parçalanmış bir ipucu var, birden fazla fiber, etc.After var, her elektrot teftiş karton sahibinin onları geri geri vermedi diseksiyon microscope.Discard elektrotlar altında incelemek her elektrot oven.Electrodes genellikle fabrikasyon sonra yaklaşık 1 ay için iyi içine. </li></ol> Bölüm 6: Karbon Fiber Kesme <ol><li> Yeri karbon fiber bir teflon bant ile cam slayt üzerine dikkatle elektrodudur. </li><li> Diseksiyon mikroskobu altında ucu gözünüzde canlandırın. </li><li> Bir hücre karşı karbon fiber yerleştirme için temiz düz bir yüzey bırakmak için karbon fiber boyunca dik kesmek için temiz bir neşter bıçak kullanın. </li></ol>

<ol>
	<li>Mundorf, M. L., Wightman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amperometry+and+cyclic+voltammetry+with+carbon+fiber+microelectrodes+at+single+cells.">Amperometry and cyclic voltammetry with carbon fiber microelectrodes at single cells.</a> Curr Protocols in Neurosci. 6 (14), 1-6 (2002).</li><li>Machado, D. J., Montesinos, M. S., Borges, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Good+practices+in+single-cell+amperometry.">Good practices in single-cell amperometry.</a> Methods in Molec. Biol. 440, 297-313 (2008).</li></ol>

Bu protokol amperometrik elektrotlar oluşturmak ve nasıl kesileceği açıklanmıştır. Amperometrik kayıtları sırasında, karbon fiber bir elektrot bir salgı hücre yüzeyine karşı yerleştirilir. Exocytotic faaliyet katekolamin oksidasyonu sonra elektron transferi ile oluşan elektrokimyasal akım gösterir amperometrik ani olarak görülmektedir. Pratik yapma ve kesme amperometrik elektrotlar sayesinde, piyasada bulunan elektrotlar pek çok üzerinde gürültü seviyeleri büyük ölçüde geliştirilmiş ol...

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Epoxy Hardener 1,3-phenylene-diamene flakes</td>
<td>Reagent</td>
<td>Sigma Aldrich</td>
<td>P23954-100G04 716AJ</td>
<td>Aliquots are stored in a dessicator and should not be exposed to light.</td>
</tr>
<tr>
<td>Carbon Fibers</td>
<td>Material</td>
<td>Goodfellow</td>
<td>C005725</td>
<td>&nbsp;</td>
<tr>
<td>Micro-Hematocrit Capillary Tubes</td>
<td>Material</td>
<td>Fisher</td>
<td>NC9836768</td>
<td>&nbsp;</td>
<tr>
<td>Epoxy Resin</td>
<td>Reagent</td>
<td>Miller-Stephenson</td>
<td>Epon Resin 828</td>
<td>&nbsp;</td>
<tr>
<td>Dissecting Microscope</td>
<td>Instrument</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Electrode Puller</td>
<td>Instrument</td>
<td>Narishige Japan</td>
<td>PP-830</td>
<td>1st pull = 24.8 degrees C; 2nd pull = 62.7 degrees C</td>
</tr>
<tr>
<td>Oven</td>
<td>Instrument</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>set to 100 degrees F.</td>
</tr>
<tr>
<td>Hot plate</td>
<td>Instrument</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>set to 55 degrees C</td>
</tr>		</tbody>
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fabrication of amperometric electrodes

Karbon fiber elektrotlar amperometri ölçümler için tek hücre veziküller katekolamin salınımının tespiti için çok önemlidir. Burada, düşük gürültü (&lt;0,5 pA) elektrotlar üretmek için gerekli teknikler açıklanmaktadır. Teknikleri önceki araştırmacıların (1,2) tarafından yayınlanan açıklamaları modifiye edilmiştir. Elektrotlar karbon elyaf hazırlama ve lif aspire bir filtre ile vakum kullanarak her kapiller tüpe ayrı ayrı iş parçacığı tarafından yapılır. Sonra, fiber ile kılcal tüpü ince sivri ucu ile iki yarısı, her bir elektrot çektirmenin tarafından çekilir. Elektrotlar sıcak, sıvı epoksi, epoksi-cam bir mühür oluşturmak için sertleştirici ile karışık batırılmış. Son olarak, epoksi kür için bir fırın elektrotlar yerleştirilir. Elektrotlar dikkatli kullanımı, tutarlı bir şekilde ve zarar vermeden yapılır sağlamak için kritik öneme sahiptir. Bu protokol tek hücre kayıt için amperometrik elektrotlar imal ve nasıl kesileceğini gösterir.

Watch this Scientific Journal Video about Fabrication of Amperometric Electrodes at JoVE.com

Fabrication of Amperometric Electrodes

Bu protokol, karbon fiber elektrotlar oluşturmak anlatılmaktadır. Bu elektrotlar daha sonra, karbon fiber amperometri ile veziküller katekolamin sürüm algılamak için kullanılır.

Amperometrik Elektrotlar imalatı

Carbon fiber electrodes are crucial for the detection of catecholamine release from vesicles in single cells for amperometry measurements.  Here, we ...

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14.3K Görüntüleme.  Saint Louis University School of Medicine. Bu protokol, karbon fiber elektrotlar oluşturmak anlatılmaktadır. Bu elektrotlar daha sonra, karbon fiber amperometri ile veziküller katekolamin sürüm algılamak için kullanılır.

Video: Fabrication of Amperometric Electrodes

Application of Light-cured Dental Adhesive Resin for Mounting Electrodes or Microdialysis Probes in Chronic Experiments

In chronic recording experiments, self-curing dental acrylic resins have been used as a mounting base of electrodes or microdialysis-probes. Since these acrylics do not bond to the bone, screws have been used as anchors. However, in small experimental animals like finches or mouse, their craniums are very fragile and can not successfully hold the anchors. In this report, we propose a new application of light-curing dental resins for mounting base of electrodes or microdialysis probes in chronic experiments. This material allows direct bonding to the cranium. Therefore, anchor screws are not required and surgical field can be reduced considerably. Past experiences show that the bonding effect maintains more than 2 months. Conventional resin's window of time when the materials are pliable and workable is a few minutes. However, the window of working time for these dental adhesives is significantly wider and adjustable.

In this report, we propose a new application of light-curing dental resins for mounting base of electrodes or microdialysis probes in chronic experiments. This material allows direct bonding to the cranium.

Kronik Deneyleri Montaj Elektrotlar veya mikrodiyaliz Problar Işık tedavi Diş Yapıştırıcı Reçine Uygulama

In chronic recording experiments, self-curing dental acrylic resins have been used as a mounting base of electrodes or microdialysis-probes. Since these ...

Biyoloji

Patterning Cells on Optically Transparent Indium Tin Oxide Electrodes

The ability to exercise precise spatial and temporal control over cell-surface interactions is an important prerequisite to the assembly of multi-cellular constructs serving as in vitro mimics of native tissues.  In this study, photolithography and wet etching techniques were used to fabricate individually addressable indium tin oxide (ITO) electrodes on glass substrates.  The glass substrates containing ITO microelectrodes were modified with poly(ethylene glycol) (PEG) silane to make them protein and cell resistive.  Presence of insulating PEG molecules on the electrode surface was verified by cyclic voltammetry employing potassium ferricyanide as a redox reporter molecule.  Importantly, the application of reductive potential caused desorption of the PEG layer, resulting in regeneration of the conductive electrode surface and appearance of typical ferricyanide redox peaks.    Application of reductive potential also corresponded to switching of ITO electrode properties from cell non-adhesive to cell-adhesive. Electrochemical stripping of PEG-silane layer from ITO microelectrodes allowed for cell adhesion to take place in a spatially defined fashion, with cellular patterns corresponding closely to electrode patterns.  Micropatterning of several cell types was demonstrated on these substrates.   In the future, the control of the biointerfacial properties afforded by this method will allow to engineer cellular microenvironments through the assembly of three or more cell types into a precise geometric configuration on an optically transparent substrate.

Non-fouling PEG silane monolayer was desorbed from individually addressable ITO electrodes on glass by application of a reductive potential. Electrochemical stripping of PEG-silane layer from ITO microelectrodes allowed for cell adhesion to take place in a spatially defined fashion, with cellular patterns corresponding closely to electrode patterns.

Optik Şeffaf indiyum kalay oksit Elektrotlar Desenlendirme Hücreler

The ability to exercise precise spatial and temporal control over cell-surface interactions is an important prerequisite to the assembly of multi-cellular ...

Fabrication of a Microfluidic Device for the Compartmentalization of Neuron Soma and Axons

In this video, we demonstrate the technique of soft lithography with polydimethyl siloxane (PDMS) which we use to fabricate a microfluidic device for culturing neurons. Previously, a silicon wafer was patterned with the design for the neuron microfluidic device using SU-8 and photolithography to create a master mold, or what we simply refer to as a "master". Next, we pour the silicon polymer PDMS on top of the master which is then cured by heating the PDMS to 80&deg;C for 1 hour. The PDMS forms a negative mold of the device. The PDMS is then carefully cut and lifted away from the master. Holes are punched where the reservoirs will be and the excess PDMS trimmed away from the device. Nitrogen is used to blow away any excess debris from the device. At this point the devices are now ready for use and can either bonded to corning No. 1 cover glass with a plasma sterilizer/cleaner or can be reversibly bound to the cover glass by simply placing the device on top of the cover glass. The reversible bonding of the device to glass is covered in a separate video and requires first that the device be sterilized either with 70% ethanol or by autoclaving. Plasma treating sterilizes the devices so no further treatment is necessary. It is, however, important, when plasma-treating the devices, to add liquid to the devices within 10 minutes of the plasma treatment while the surfaces are still hydrophilic. Waiting longer than 10 minutes to add liquid to the device makes it difficult for the liquid to enter the device. The neuron devices are typically plasma-bound to cover glass and 0.5 mg/ml poly-L-lysine (PLL) in pH 8.5 borate buffer is immediately added to the device. After a minimum of 3 hours incubating with PLL, the devices are washed with dH2O water a minimum of 3 times with at least 15 minutes between each wash. Next, the water is removed and fresh media is added to the device. At this point the device is ready for use. It is important to remember at this point to never remove all the media from the device. Always leave media in the main channel.

In this video we demonstrate the technique of soft lithography with polydimethyl siloxane (PDMS) which we use to farbricate a microfluidic device for culturing neurons.

Nöron Soma ve Aksonlar compartmentalization mikroakışkan Aygıt Fabrikasyon

In this video, we demonstrate the technique of soft lithography with polydimethyl siloxane (PDMS) which we use to fabricate a microfluidic device for ...

PDMS Device Fabrication and Surface Modification

PDMS Cihaz İmalat ve Yüzey Modifikasyonu

Non-plasma Bonding of PDMS for Inexpensive Fabrication of Microfluidic Devices

In this video, we demonstrate how to use the neuron microfluidic device without plasma bonding.  In some cases it may be desirable to reversibly bond devices to the Corning No. 1 cover glass.  This could be due, perhaps, to a plasma cleaner not being available.  In other instances, it may be desirable to remove the device from the glass after the culturing of neurons for certain types of microscopy or for immunostaining, though it is not necessary to remove the device for immunostaining since the neurons can be stained in the device.  Some researchers, however, still prefer to remove the device.  In this case, reversible bonding of the device to the cover glass makes that possible.  There are some disadvantages to non-plasma bonding of the devices in that not as tight of a seal is formed.  In some cases axons may grow under the grooves rather than through them.  Also, because the glass and PDMS are hydrophobic, liquids do not readily enter the device making it necessary at times to force media and other reagents into the device.  Liquids will enter the device via capillary action, but it takes significantly longer as compared to devices that have been plasma bonded.  The plasma cleaner creates temporary hydrophilic charges on the glass and device that facilitate the flow of liquids through the device after bonding within seconds.  For non-plasma bound devices, liquid flow through the devices takes several minutes.  It is also important to note that the devices to be used with non-plasma bonding need to be sterilized first, whereas plasma treated devices do not need to be sterilized prior to use because the plasma cleaner will sterilize them.

In this video we demonstrate how to use the neuron microfluidic device without plasma bonding.

Ucuz mikroakışkan Cihazlar İmalat PDMS Non-plazma Yapışma

In this video, we demonstrate how to use the neuron microfluidic device without plasma bonding.  In some cases it may be desirable to reversibly bond ...

Fabrication of the Thermoplastic Microfluidic Channels

In our lab, we have successfully isolated nucleic acids directly from microliter and submicroliter volumes of human blood, urine and stool using polymer/nanoparticle composite microscale lysis and solid phase extraction columns. The recovered samples are concentrated, small volume samples that are PCRable, without any additional cleanup. Here, we demonstrate how to fabricate thermoplastic microfluidic chips using hot embossing and heat sealing. Then, we demonstrate how to use in situ light directed surface grafting and polymerization through the sealed chip to form the composite solid phase columns. We demonstrate grafting and polymerization of a carbon nanotube/polymer composite column for bacterial cell lysis. We then show the lysis process followed by solid phase extraction of nucleic acids from the sample on chip using a silica/polymer composite column. The attached protocols contain detailed instructions on how to make both lysis and solid phase extraction columns.

Here we demonstrate how to fabricate thermoplastic microfluidic chips using hot embossing and heat sealing. Then we demonstrate how to use in situ light directed surface grafting and polymerization through the sealed chip to form the composite solid phase columns.

Termoplastik mikroakışkan Kanallar imalatı

In our lab, we have successfully isolated nucleic acids directly from microliter and submicroliter volumes of human blood, urine and stool using ...

Making Patch-pipettes and Sharp Electrodes with a Programmable Puller

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades. Today, such pipettes are made from glass capillaries using a programmable puller. Such instruments heat the capillary using either a metal filament or a laser and draw out the glass using gravity, a motor or both. Pipettes for patch-clamp recording are formed using only heat and gravity, while sharp electrodes for intracellular recording use a combination of heat, gravity, and a motor. The procedure used to make intracellular recording pipettes is similar to that used to make injection needles for a variety of applications, including cRNA injection into Xenopus oocytes. In general, capillary glass &lt;1.2 mm in diameter is used to make pipettes for patch clamp recording, while narrower glass is used for intracellular recording (outer diameter = 1.0 mm). For each tool, the puller is programmed slightly differently. This video shows how to make both kinds of recording pipettes using pre-established puller programs.

This video shows how to use a programmable puller to make patch pipettes and sharp electrodes for electrophysiology. The same procedure can be used to make a variety of glass tools, including injection needles.

Programlanabilir Çektirme Patch-pipetler ve Sharp Elektrotlar

Glass microelectrodes (also called pipettes) have been a workhorse of electrophysiology for decades.  Today, such pipettes are made from glass capillaries ...

Fabrication of Myogenic Engineered Tissue Constructs

Despite the fact that electronic pacemakers are life-saving medical devices, their long-term performance in pediatric patients can be problematic owing to the restrictions imposed by a child's small size and their inevitable growth. Consequently, there is a genuine need for innovative therapies designed specifically for pediatric patients with cardiac rhythm disorders. We propose that a conductive biological alternative consisting of a collagen-based matrix containing autologously-derived cells could better adapt to growth, reduce the need for recurrent surgeries, and greatly improve the quality of life for these patients. In the present study, we describe a procedure for incorporating primary skeletal myoblast cell cultures within a hydrogel matrix to fashion a surgically-implantable tissue construct that will serve as an electrical conduit between the upper and lower chambers of the heart. Ultimately, we anticipate using this type of engineered tissue to restore atrioventricular electrical conduction in children with complete heart block. In view of that, we isolate myoblasts from the skeletal muscles of neonatal Lewis rats and plate them onto laminin-coated tissue culture dishes using a modified version of established protocols[2, 3]. After one to two days, cultured cells are collected and mixed with antibiotics, type 1 collagen, Matrigel&trade;, and NaHCO3. The result is a viscous, uniform solution that can be cast into a mold of nearly any shape and size[1, 4, 5]. For our tissue constructs, we employ type 1 collagen isolated from fetal lamb skin using standard procedures[6]. Once the tissue has solidified at 37&deg;C, culture media is carefully added to the plate until the construct is submerged. The engineered tissue is then allowed to further condense through dehydration for 2 more days, at which point it is ready for in vitro assessment or surgical-implantation.

Here, we demonstrate fabrication of collagen-based, tissue constructs containing skeletal myoblasts. These 3-D engineered constructs may be used to replace or repair tissues in vivo. For our purposes, we have designed these as an atrioventricular electrical conduit for the repair of complete heart block[1].

MİYOJEN Engineered Doku oluşturur imalatı

Despite the fact that electronic pacemakers are life-saving medical devices, their long-term performance in pediatric patients can be problematic owing to ...

Fabrication and Operation of an Oxygen Insert for Adherent Cellular Cultures 

Oxygen is a key modulator of many cellular pathways, but current devices permitting in vitro oxygen modulation fail to meet the needs of biomedical research. The hypoxic chamber offers a simple system to control oxygenation in standard culture vessels, but lacks precise temporal and spatial control over the oxygen concentration at the cell surface, preventing its application in studying a variety of physiological phenomena. Other systems have improved upon the hypoxic chamber, but require specialized knowledge and equipment for their operation, making them intimidating for the average researcher. A microfabricated insert for multiwell plates has been developed to more effectively control the temporal and spatial oxygen concentration to better model physiological phenomena found in vivo. The platform consists of a polydimethylsiloxane insert that nests into a standard multiwell plate and serves as a passive microfluidic gas network with a gas-permeable membrane aimed to modulate oxygen delivery to adherent cells. The device is simple to use and is connected to gas cylinders that provide the pressure to introduce the desired oxygen concentration into the platform. Fabrication involves a combination of standard SU-8 photolithography, replica molding, and defined PDMS spinning on a silicon wafer. The components of the device are bonded after surface treatment using a hand-held plasma system. Validation is accomplished with a planar fluorescent oxygen sensor. Equilibration time is on the order of minutes and a wide variety of oxygen profiles can be attained based on the device design, such as the cyclic profile achieved in this study, and even oxygen gradients to mimic those found in vivo. The device can be sterilized for cell culture using common methods without loss of function. The device's applicability to studying the in vitro wound healing response will be demonstrated.

Fabrication and Operation of an Oxygen Insert for Adherent Cellular Cultures

Fabrication and validation of an add-on platform that offers enhanced control over the spatial and temporal oxygenation in a 6-well plate. The device is adaptable to a number of culture systems and can be used to investigate the effects of oxygen on wound healing.

İmalat ve Operasyon yapışkan Hücresel Kültürler Oksijen Ekle

Oxygen is a key modulator of many cellular pathways, but current devices permitting in vitro oxygen modulation fail to meet the needs of biomedical ...

Fabrication and Use of MicroEnvironment microArrays (MEArrays)

The interactions between cells and their surrounding microenvironment have functional consequences for cellular behaviour. On the single cell level, distinct microenvironments can impose differentiation, migration, and proliferation phenotypes, and on the tissue level the microenvironment processes as complex as morphogenesis and tumorigenesis1. Not only do the cell and molecular contents of microenvironments impact the cells within, but so do the elasticity2 and geometry3 of the tissue. Defined as the sum total of cell-cell, -ECM, and -soluble factor interactions, in addition to physical characteristics, the microenvironment is complex. The phenotypes of cells within a tissue are partially due to their genomic content and partially due to the combinatorial interactions with the microenviroment. A major challenge is to link specific combinations of microenvironmental components with distinctive behaviours.
Here, we present the microenvironment microarray (MEArray) platform for cell-based functional screening of interactions with combinatorial microenvironments4. The method allows for simultaneous control of the molecular composition and the elastic modulus, and combines the use of widely available microarray and micropatterning technologies. MEArray screens require as few as 10,000 cells per array, which facilitates functional studies of rare cell types such as adult progenitor cells. A limitation of the technology is that entire tissue microenvironments cannot be completely recapitulated on MEArrays. However, comparison of responses in the same cell type to numerous related microenvironments, for instance pairwise combinations of ECM proteins that characterize a given tissue, will provide insights into how microenvironmental components elicit tissue-specific functional phenotypes.
MEArrays can be printed using a wide variety of recombinant growth factors, cytokines, and purified ECM proteins, and combinations thereof. The platform is limited only by the availability of specific reagents. MEArrays are amenable to time-lapsed analysis, but most often are used for end point analyses of cellular functions that are measureable with fluorescent probes. For instance, DNA synthesis, apoptosis, acquisition of differentiated states, or production of specific gene products are commonly measured. Briefly, the basic flow of an MEArray experiment is to prepare slides coated with printing substrata and to prepare the master plate of proteins that are to be printed. Then the arrays are printed with a microarray robot, cells are allowed to attach, grow in culture, and then are chemically fixed upon reaching the experimental endpoint. Fluorescent or colorimetric assays, imaged with traditional microscopes or microarray scanners, are used to reveal relevant molecular and cellular phenotypes (Figure 1).

Fabrication and Use of MicroEnvironment microArrays MEArrays

A combinatorial functional screening method for gaining insights into the impacts of the molecular composition of microenvironments on cellular functions is described. The method takes advantage of existing microarray-based technologies to generate arrays of defined combinatorial microenvironments that support cell adhesion and functional analysis.

Fabrikasyon ve mikroçevresindeki microarrays Kullanımı (MEArrays)

The interactions between cells and their surrounding microenvironment have functional consequences for cellular behaviour. On the single cell level, ...