Name: tracking drug-induced changes in receptor post-internalization trafficking by colocalizational analysis
Uploaded: 2015-07-03T15:00:00
Description: Watch this Scientific Journal Video about Tracking Drug-induced Changes in Receptor Post-internalization Trafficking by Colocalizational Analysis at JoVE.com

Dette arbejde blev støttet af en bevilling fra CIHR (MOP394808) og en Canada Research Chair til CMCEWO var modtageren af ​​en Post-Graduate Scholarship fra NSERC.

Bemærk: Denne protokol er i store træk forenelig med forskellige monolag-belagte celle / vævskultur modeller, narkotika behandlingsregimer og mærkning mål. Således i den faktiske brug, vil mange specifikke parametre variere baseret på eksperimentelle design. Her henvisninger til disse brugerdefinerede parametre er generiske. Eksempelvis betingelser, som anvendes til at opnå de repræsentative resultater, er anført i kursiv. 1. Opløsninger <ol><li> Forbered vaskebuffer ved blandin...

<ol>
	<li>Drake, M. T., Shenoy, S. K., Lefkowitz, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trafficking+of+G+protein-coupled+receptors.">Trafficking of G protein-coupled receptors.</a> Circ. Res. 99 (6), 570-582 (2006).</li><li>Hanyaloglu, A. C., von Zastrow, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+GPCRs+by+endocytic+membrane+trafficking+and+its+potential+implications.">Regulation of GPCRs by endocytic membrane trafficking and its potential implications.</a> Annu. Rev. Pharmacol. Toxicol. 48, 537-568 (2008).</li><li>Hislop, J. 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L., Enquist, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+postendocytic+sorting+of+G+protein-coupled+receptors.">Modulation of postendocytic sorting of G protein-coupled receptors.</a> Science. 297 (5581), 615-620 (2002).</li><li>Von Zastrow, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+opioid+receptors+by+endocytic+membrane+traffic:+mechanisms+and+translational+implications.">Regulation of opioid receptors by endocytic membrane traffic: mechanisms and translational implications.</a> Drug Alcohol Depend. 108 (3), 166-171 (2010).</li><li>Milan-Lobo, L., Whistler, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heteromerization+of+the+&#181;-+and+&#948;-opioid+receptors+produces+ligand-biased+antagonism+and+alters+&#181;-receptor+trafficking.">Heteromerization of the &#181;- and &#948;-opioid receptors produces ligand-biased antagonism and alters &#181;-receptor trafficking.</a> J. Pharmacol. Exp. Ther. 337 (3), 868-875 (2011).</li><li>Wang, H. -. B., Guan, J. -. S., Bao, L., Zhang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+subcellular+distribution+of+delta-opioid+receptor+fused+with+various+tags+in+PC12+cells.">Distinct subcellular distribution of delta-opioid receptor fused with various tags in PC12 cells.</a> Neurochem. Res. 33 (10), 2028-2034 (2008).</li><li>Scherrer, G., Tryoen-T&#243;th, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Knockin+mice+expressing+fluorescent+delta-opioid+receptors+uncover+G+protein-coupled+receptor+dynamics+in+vivo.">Knockin mice expressing fluorescent delta-opioid receptors uncover G protein-coupled receptor dynamics in vivo.</a> Proc. Natl. Acad. Sci. 103 (25), 9691-9696 (2006).</li><li>He, S. -. Q., Zhang, Z. -. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Facilitation+of+&#181;-opioid+receptor+activity+by+preventing+&#948;-opioid+receptor-mediated+codegradation.">Facilitation of &#181;-opioid receptor activity by preventing &#948;-opioid receptor-mediated codegradation.</a> Neuron. 69 (1), 120-131 (2011).</li><li>French, A. P., Mills, S., Swarup, R., Bennett, M. J., Pridmore, T. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Colocalization+of+fluorescent+markers+in+confocal+microscope+images+of+plant+cells.">Colocalization of fluorescent markers in confocal microscope images of plant cells.</a> Nat. Protoc. 3 (4), 619-628 (2008).</li><li>Ong, E. W., Xue, L., Olmstead, M. C., Cahill, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prolonged+morphine+treatment+alters+&#948;+opioid+receptor+post-internalization+trafficking.">Prolonged morphine treatment alters &#948; opioid receptor post-internalization trafficking.</a> Br. J. Pharmacol. 172 (2), 615-629 (2014).</li><li>Schnell, U., Dijk, F., Sjollema, K. a. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunolabeling+artifacts+and+the+need+for+live-cell+imaging.">Immunolabeling artifacts and the need for live-cell imaging.</a> Nature Methods. 9 (2), 152-158 (2012).</li><li>Rozenfeld, R., Gupta, A., Devi, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+surface+targeting+of+mu-delta+opioid+receptor+heterodimers+by+RTP4.">Cell surface targeting of mu-delta opioid receptor heterodimers by RTP4.</a> Proc. Natl. Acad. Sci. 105 (41), 16045-16050 (2008).</li><li>Gupta, A., Mulder, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+abundance+of+opioid+receptor+heteromers+after+chronic+morphine+administration.">Increased abundance of opioid receptor heteromers after chronic morphine administration.</a> Sci. Signal. 3 (131), ra54 (2010).</li><li>Seibenhener, M. L., Wooten, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+culture+of+hippocampal+neurons+from+prenatal+mice.">Isolation and culture of hippocampal neurons from prenatal mice.</a> J. Vis. Exp. (65), 1-7 (2012).</li><li>Bolte, S., Cordeli&#232;res, F. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+guided+tour+into+subcellular+colocalization+analysis+in+light+microscopy.">A guided tour into subcellular colocalization analysis in light microscopy.</a> J. Microsc. 224 (Pt 3), 213-232 (2006).</li><li>Field, A., Miles, J., Field, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Discovering Statistics Using R. , (2012).</li><li>Mattioli, T. -. A. M., Milne, B., Cahill, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultra-low+dose+naltrexone+attenuates+chronic+morphine-induced+gliosis+in+rats.">Ultra-low dose naltrexone attenuates chronic morphine-induced gliosis in rats.</a> Mol. Pain. 6, 22 (2010).</li><li>Vogt, R. F., Phillips, D. L., Henderson, L. O., Whitfield, W., Spierto, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+differences+among+various+proteins+as+blocking+agents+for+ELISA+microtiter+plates.">Quantitative differences among various proteins as blocking agents for ELISA microtiter plates.</a> J. Immunol. Methods. 101 (1), 43-50 (1987).</li><li>Johnson, I., Spence, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Molecular Probes Handbook. , (2010).</li><li>Morris, T. J., Stanley, E. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+for+immunocytochemical+staining+with+multiple+rabbit+polyclonal+antibodies.">A simple method for immunocytochemical staining with multiple rabbit polyclonal antibodies.</a> J. Neurosci. Methods. 127 (2), 149-155 (2003).</li><li>Pawley, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Handbook of Biological Confocal Microscopy. , (2006).</li><li>North, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Seeing+is+believing?+A+beginners&#8217;+guide+to+practical+pitfalls+in+image+acquisition.">Seeing is believing? A beginners&#8217; guide to practical pitfalls in image acquisition.</a> J. Cell Biol. 172 (1), 9-18 (2006).</li><li>Zinchuk, V., Zinchuk, O., Okada, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+colocalization+analysis+of+multicolor+confocal+immunofluorescence+microscopy+images:+pushing+pixels+to+explore+biological+phenomena.">Quantitative colocalization analysis of multicolor confocal immunofluorescence microscopy images: pushing pixels to explore biological phenomena.</a> Acta Histochem. Cytochem. 40 (4), 101-111 (2007).</li></ol>

Vi har optimeret denne protokol til analyse af primære kulturer af voksne dorsale ganglie-neuroner (primære sensoriske neuroner). Den kan også anvendes, med ringe eller ingen ændring, for monolag-belagte dyrkede celler bredt. Den colocalizational analyse er også muligt i vævssnit og andre sådanne præparater 11, men komponenterne narkotikabehandling og vævsfiksering / forberedelse ville ikke være hensigtsmæssigt. Af interesse, kan også anvendes ICC metoder præsenteres ...

Receptorer, især G-protein koblede receptorer (GPCR&#39;er), rutinemæssigt trafikerede intracellulært til og fra celleoverfladen 1. Disse komplekst orkestreret og stramt styrede processer diktere cellers tilgængelige receptor supplerer og regulere receptor tidsmæssig aktivitet, desensibilisering, og resensitization 2 - 4. Vigtigere er disse processer reagerer på cellulære miljøer, herunder lægemiddelinduceret receptor aktivitet eller inaktivitet. Det vil sige, kan handlinger ligander på receptorer ændre intracellulær handel med disse receptorer, og derved ændre celle lydhørhed. På denne måde eksterne ligander udøver endnu flere effekter på celle funktion, også ud over klassiske messenger-til-effektor kaskader 5,6. Undersøgelse sådanne ændringer i induceret receptor handel er vanskelig. Alle tilgængelige teknikker indebærer begrænsninger. Biotin beskyttelse assays er blevet anvendt til at monitor overfladereceptorer befolkninger. Disse receptorer er biotinyleret og et tidsforløb af immunopræcipitationer udføres for at kvantificere reduktionen i biotinylerede receptorer over tid. Denne teknik væsentlige overvåger den gradvise nedbrydning af en indledende, mærket, population af receptorer 7, og er meget nyttig ved konstruktion tidsforløb for denne proces. Desværre dette assay er i stand til at overvåge enhver anden end nedbrydning af den oprindelige pulje af receptorer, såsom internalisering, genvinding eller nye receptorer proces. Desuden kan tilsætningen af et antistof i 150kDa interval til en receptor i 50 kDa området ændre receptorens handel 8,9, og denne teknik kan være vanskeligt at bruge med lave ekspression niveau receptorer. Andre procedurer bruger forskellige metoder til at identificere rum intracellulære menneskehandel (f.eks endosomer, etc.), og vurdere deres colokalisering med receptorerne af interesse. Dette omfatter ose af heterologe systemer udtrykker fluorescerende-protein-mærkede kimære konstruktioner af receptorerne og compartment markører (f.eks Rab-familie GTPaser). Dette gør det muligt potentielt brugen af ​​live-cell imaging, fjerne spørgsmål vedrørende fiksering og permeabilisering. Mens kraftfuld, sådan strategi lider under de samme begrænsninger for heterologe systemer generelt: tag og udtryk niveau effekter på menneskehandel adfærd og uforenelighed med mere fysiologisk repræsentative celletyper. Mere populært, er farvestoffer bruges til nemt mærke intracellulære rum (f.eks lysosomer, angiveligt) 10. Farvestoffer kan imidlertid mangle specificitet (alle sure organeller i tilfælde af farvestoffer til lysosomer) og ikke vurderer menneskehandel gennem andre rum. Alligevel disse teknikker tillader betydelig kontrol over systemet og eksperimentelle betingelser, og kan drage fordel af de colokalisering analysemetoder præsenteres her nedenfor. Fremgangsmåden we til stede her forfiner sporing af receptor menneskehandel ved colokalisering. Anvendelse af immuncytokemi (ICC) til at mærke passende markører, er det muligt at identificere flere distinkte intracellulære rum. Dette tillader også anvendelse af fysiologisk relevante primære cellekulturer i stedet for heterologe systemer. Denne ICC-protokollen indebærer fastsættelse cellerne af interesse forud for mærkning; dette tillader mærkning på et bestemt tidspunkterne efter lægemiddelbehandling (er). Dette giver en &quot;øjebliksbillede&quot; af de globale receptor-rum foreninger på det tidspunkterne. Med forskellige tidspunkter, kan et tidsforløb af ændringer trafficking også konstrueres. Kort fortalt celler er lægemiddelbehandlede, mærket for receptoren og intracellulære rum af interesse, konfokalt afbildes, og mikrofotografierne analyseres matematisk kvantificere colokalisering af receptoren og rummet 11. I vores brug undersøgte vi colokalisering af en receptor med Rab5, Rab11 og Lysosomal-associeret membranprotein 1 (LAMP1). Disse markører identificere tidlige endosomer, genbrug endosomer og lysosomer henholdsvis. Disse co-lokaliseringstoppe foranstaltninger fungerer som stedfortrædere for de overordnede processer i internalisering, genbrug og nedbrydning 12. Som med alle teknikker, bør visse begrænsninger overvejes. På grund af behovet for at billedet hver enkelt neuron analyseres, kan denne teknik blive ganske arbejdskrævende afhængigt af antallet af betingelser og tidspunkter involveret. Alle immunolabeling skal også kæmpe med virkningerne på cellulært ultrastruktur, protein lokalisering og epitop tilgængelighed forårsaget af fiksering og permeabilisering 13. Selvom oprindeligt optimeret til brug med primære kulturer af primære sensoriske neuroner, er denne fremgangsmåde stort set forenelig med andre kulturmodeller monolag-belagt. Anvendelsen af ​​en matematisk kvantificeret measure af colokalisering er, især langt mere metodisk stringent end tidligere teknikker, der anvendes til at vurdere ændringer receptor menneskehandel, som ofte påberåbes vage, subjektive foranstaltninger såsom visuelt inspicerede multi-kanal overlays 14. Denne teknik er særlig nyttig for sin brede kompatibilitet med in vivo interventioner (før primær generation kultur), in vitro interventioner (under væksten kultur) og diverse mærkning rettet mod 15. Som sådan kan den tilpasses til mange forskellige forskningsspørgsmål.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Trizma Base</td>
			<td>Sigma Aldrich</td>
			<td>T1503-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Chloride</td>
			<td>Sigma Aldrich</td>
			<td>S9888-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween 20</td>
			<td>Fisher Scientific</td>
			<td>BP337-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrochloric Acid</td>
			<td>Sigma Aldrich</td>
			<td align="right">258148</td>
			<td></td>
		</tr>
		<tr>
			<td>Albumin from Bovine Serum</td>
			<td>Sigma Aldrich</td>
			<td>A7906-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelatin from cold water fish skin</td>
			<td>Sigma Aldrich</td>
			<td>G7041-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Corning Costar Cell Culture Plates: 24-well</td>
			<td>Fisher Scientific</td>
			<td align="right">720084</td>
			<td></td>
		</tr>
		<tr>
			<td>12 circle Microscope Cover Glass</td>
			<td>Fisher Scientific</td>
			<td align="right">1254580</td>
			<td></td>
		</tr>
		<tr>
			<td>Aqua/Poly-Mount</td>
			<td>Polysciences</td>
			<td>18606-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Phosphate Monobasic</td>
			<td>Sigma Aldrich</td>
			<td>S9638-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Phosphate Dibasic</td>
			<td>Sigma Aldrich</td>
			<td>S9763-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Polysciences</td>
			<td>00380-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #5 Forceps - Standard/Dumoxel</td>
			<td>Fine Science Tools</td>
			<td>11251-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Rabbit anti-DOR antibody</td>
			<td>MyBioSource</td>
			<td>MBS316175</td>
			<td>Used at 1:1500</td>
		</tr>
		<tr>
			<td>Mouse anti-Rab5 antibody</td>
			<td>Sigma Aldrich</td>
			<td>R7904</td>
			<td>Used at 1:750</td>
		</tr>
		<tr>
			<td>Mouse anti-Rab11 antibody</td>
			<td>Millipore</td>
			<td>05-853</td>
			<td>Used at 1:500</td>
		</tr>
		<tr>
			<td>Goat anti-LAMP1 antibody</td>
			<td>Santa Cruz</td>
			<td>SC8098</td>
			<td>Used at 1:750</td>
		</tr>
		<tr>
			<td>Donkey anti-rabbit Alexa 488 conjugated antibody</td>
			<td>Life Technologies</td>
			<td>A-21206</td>
			<td>Used at 1:200 to 1:2000</td>
		</tr>
		<tr>
			<td>Goat anti-mouse Alexa 594 conjugated antibody</td>
			<td>Life Technologies</td>
			<td>A-11005</td>
			<td>Used at 1:200 to 1:2000</td>
		</tr>
		<tr>
			<td>Donkey anti-goat Alexa 594 conjugated antibody</td>
			<td>Life Technologies</td>
			<td>A-11058</td>
			<td>Used at 1:200 to 1:2000</td>
		</tr>
	</tbody>
</table>

tracking drug-induced changes in receptor post-internalization trafficking by colocalizational analysis

The intracellular trafficking of receptors is a collection of complex and highly controlled processes. Receptor trafficking modulates signaling and overall cell responsiveness to ligands and is, itself, influenced by intra- and extracellular conditions, including ligand-induced signaling. Optimized for use with monolayer-plated cultured cells, but extendable to free-floating tissue slices, this protocol uses immunolabelling and colocalizational analysis to track changes in intracellular receptor trafficking following both chronic/prolonged and acute interventions, including exogenous drug treatment. After drug treatment, cells are double-immunolabelled for the receptor and for markers for the intracellular compartments of interest. Sequential confocal microscopy is then used to capture two-channel photomicrographs of individual cells, which are subjected to computerized colocalizational analysis to yield quantitative colocalization scores. These scores are normalized to permit pooling of independent replicates prior to statistical analysis. Representative photomicrographs may also be processed to generate illustrative figures. Here, we describe a powerful and flexible technique for quantitatively assessing induced receptor trafficking.

Med denne teknik er det muligt at kvantificere ændringer i receptor efter internalisering trafficking følgende både kroniske / langvarig og akutte medicinske behandlingsmetoder. Efter medicinsk behandling, fiksering og mærkning, er i høj opløsning to-kanals mikrofotografier fanget i hver celle af interesse. Repræsentative billeder kan kombineres med falske farver colokalisering at generere illustrative tal (figur 1). Efterfølgende colocalizational analyse, som beskrevet, udbytte kvantitative sne...

Watch this Scientific Journal Video about Tracking Drug-induced Changes in Receptor Post-internalization Trafficking by Colocalizational Analysis at JoVE.com

Tracking Drug-induced Changes in Receptor Post-internalization Trafficking by Colocalizational Analysis

Receptor menneskehandel modulerer signalering og celle lydhørhed over for ligander og er i sig selv reagerer på celle, herunder ligand-induceret signalering. Her beskriver vi en stærk og fleksibel teknik til kvantitativ vurdering af lægemiddelinduceret receptor menneskehandel hjælp immunolabeling og colocalizational analyse.

Sporing lægemiddelinduceret Ændringer i Receptor Post-internalisering Trafficking af Colocalizational Analysis

The intracellular trafficking of receptors is a collection of complex and highly controlled processes. Receptor trafficking modulates signaling and ...

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Analyse af SCAP<em&gt; N</em&gt; -glycosylation Og handel med humane celler

Elevated lipogenesis is a common characteristic of cancer and metabolic diseases. Sterol regulatory element-binding proteins (SREBPs), a family of ...

Live Cell Imaging and 3D Analysis of Angiotensin Receptor Type 1a Trafficking in Transfected Human Embryonic Kidney Cells Using Confocal Microscopy

Levende celler og 1a 3D Analyse af angiotensin receptor type handel i transfekterede humane embryoniske nyreceller Brug konfokalmikroskopi

Live-cell imaging is used to simultaneously capture time-lapse images of angiotensin type 1a receptors (AT1aR) and intracellular compartments in ...

Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy

Påvisning af ligand-aktiverede G-protein-koblede receptorer Internalisering af konfokal mikroskopi

Confocal laser scanning microscopy (CLSM) is an optical imaging technique for high-contrast imaging. It is a powerful approach to visualize fluorescent ...

A Graphical User Interface for Software-assisted Tracking of Protein Concentration in Dynamic Cellular Protrusions

Et grafisk brugergrænseflade til softwareassisteret sporing af proteinkoncentration i dynamiske cellulære protrusioner

Filopodia are dynamic, finger-like cellular protrusions associated with migration and cell-cell communication. In order to better understand the complex ...

Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers

Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers

Visualisering og sporing endogene mRNAs i levende Drosophila melanogaster ægge kamre

Fluorescence-based imaging techniques, in combination with developments in light microscopy, have revolutionized how cell biologists conduct live cell ...