Name: single-cell resolution fluorescence live imaging of drosophila circadian clocks in larval brain culture
Uploaded: 2018-01-19T15:00:00
Description: Watch this Scientific Journal Video about Single-cell Resolution Fluorescence Live Imaging of Drosophila Circadian Clocks in Larval Brain Culture at JoVE.com

Vi takker Michael Rosbash for hans mentorarbejde og støtte i den indledende fase af udviklingen af denne metode. Dette arbejde blev finansieret af JST PRESTO programmet, den schweiziske National Science Foundation (31003A_149893 og 31003A_169548), det Europæiske Forskningsråd (ERC-StG-311194), Novartis Foundation for medicinsk-biomedicinsk forskning (13A39) og universitetet i Genève .

1. fremstilling af stamopløsninger Under en kultur hætte
<ol><li>Forberede 400 mL 1 x Schneider Active Medium (SAM) optimeret til ex vivo kultur af larve hjerner (modificeret fra reference24,25) (tabel 1). Alikvot til 5 mL og flash fryse i flydende kvælstof (LN2), opbevares ved - 80 ° C.</li>
	<li>Forbered 1 x Dissecting saltopløsning (DSS) ved at fortynde 10 x stamopløsning (modificeret fra reference...

<ol>
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C., 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=A+G+protein-coupled+receptor,+groom-of-PDF,+is+required+for+PDF+neuron+action+in+circadian+behavior.">A G protein-coupled receptor, groom-of-PDF, is required for PDF neuron action in circadian behavior.</a> Neuron. 48 (2), 221-227 (2005).</li><li>Mertens, I., 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=PDF+receptor+signaling+in+Drosophila+contributes+to+both+circadian+and+geotactic+behaviors.">PDF receptor signaling in Drosophila contributes to both circadian and geotactic behaviors.</a> Neuron. 48 (2), 213-219 (2005).</li><li>Ayaz, D., 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=Axonal+injury+and+regeneration+in+the+adult+brain+of+Drosophila.">Axonal injury and regeneration in the adult brain of Drosophila.</a> J Neurosci. 28 (23), 6010-6021 (2008).</li><li>Prithviraj, R., Trunova, S., Giniger, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ex+vivo+culturing+of+whole,+developing+Drosophila+brains.">Ex vivo culturing of whole, developing Drosophila brains.</a> J Vis Exp. 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(37), (2010).</li><li>Roberts, L., 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=Light+evokes+rapid+circadian+network+oscillator+desynchrony+followed+by+gradual+phase+retuning+of+synchrony.">Light evokes rapid circadian network oscillator desynchrony followed by gradual phase retuning of synchrony.</a> Curr Biol. 25 (7), 858-867 (2015).</li><li>Sehadova, H., 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=Temperature+entrainment+of+Drosophila's+circadian+clock+involves+the+gene+nocte+and+signaling+from+peripheral+sensory+tissues+to+the+brain.">Temperature entrainment of Drosophila's circadian clock involves the gene nocte and signaling from peripheral sensory tissues to the brain.</a> Neuron. 64 (2), 251-266 (2009).</li><li>Sellix, M. T., Currie, J., Menaker, M., Wijnen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence/luminescence+circadian+imaging+of+complex+tissues+at+single-cell+resolution.">Fluorescence/luminescence circadian imaging of complex tissues at single-cell resolution.</a> J Biol Rhythms. 25 (3), 228-232 (2010).</li><li>Liang, X., Holy, T. E., Taghert, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synchronous+Drosophila+circadian+pacemakers+display+nonsynchronous+Ca(2)(+)+rhythms+in+vivo.">Synchronous Drosophila circadian pacemakers display nonsynchronous Ca(2)(+) rhythms in vivo.</a> Science. 351 (6276), 976-981 (2016).</li><li>Liang, X., Holy, T. E., Taghert, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Series+of+Suppressive+Signals+within+the+Drosophila+Circadian+Neural+Circuit+Generates+Sequential+Daily+Outputs.">A Series of Suppressive Signals within the Drosophila Circadian Neural Circuit Generates Sequential Daily Outputs.</a> Neuron. , (2017).</li><li>Kuppers-Munther, B., 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=A+new+culturing+strategy+optimises+Drosophila+primary+cell+cultures+for+structural+and+functional+analyses.">A new culturing strategy optimises Drosophila primary cell cultures for structural and functional analyses.</a> Dev Biol. 269 (2), 459-478 (2004).</li><li>Schneider, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differentiation+of+Larval+Drosophila+Eye-Antennal+Discs+in+Vitro.">Differentiation of Larval Drosophila Eye-Antennal Discs in Vitro.</a> J Exp Zool. 156, 91-103 (1964).</li><li>Jiang, S. A., Campusano, J. M., Su, H., O'Dowd, D. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+mushroom+body+Kenyon+cells+generate+spontaneous+calcium+transients+mediated+by+PLTX-sensitive+calcium+channels.">Drosophila mushroom body Kenyon cells generate spontaneous calcium transients mediated by PLTX-sensitive calcium channels.</a> J Neurophysiol. 94 (1), 491-500 (2005).</li><li>Hafer, N., Schedl, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dissection+of+larval+CNS+in+Drosophila+melanogaster.">Dissection of larval CNS in Drosophila melanogaster.</a> J Vis Exp. 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Her beskrev vi metoden for langsigtede fluorescens time-lapse mikroskopi af kulturperler larve hjerner. Succes af denne slags eksperimenter, afhænger af flere faktorer, såsom kultur, sundhed metode for immobilisering af hjerne eksplantat, fluorescens intensitet og signal / støj forholdet mellem reporter, tidsmæssige og rumlige opløsning, og tilgængelighed til eksplantat. Disse faktorer kan være gensidigt udelukker hinanden. For eksempel, stigende time-lapse hyppighed påvirker sundheden for kulturen, og immobilise...

Døgnrytmen ure hjælpe organismer til at tilpasse sig de periodiske miljøforandringer genereret af 24 h rotation af jorden. Låst transcriptional-translationel feedback-sløjfer almindeligvis ligge til grund for de molekylære maskineri af døgnrytmen ure på tværs af arter1. Døgnrytmen pacemaker kredsløb bestående af ur-holdige neuroner omfatter tid af dagen oplysninger formidles af de miljømæssige stikord, som lys/mørke (LD) og temperatur cyklusser, at orkestrere rytmer i et væld af daglige fysiologiske og adfærdsmæssige processer2,3. Koordinering af Molekylær rytme med neuronal input og output er af afgørende betydning for driften af døgnrytmen kredsløb, men stadig kun delvist forstået.
I Drosophila, kernen i den molekylære ur, TAKTCYKLUS (CLK/CYC) heterodimer aktiveres transskription af periode (per) og tidløse (tim). PER og TIM udgør en kompleks og Indtast atomkernen, hvor de hæmmer transcriptional aktivitet på CLK/CYC og dermed deres egen transskription. Post-Transcriptional og posttranslationelle forordninger forårsage forsinkelser mellem CLK/CYC-medieret transskription og undertrykkelse af PER / TIM, sikre generation af circa 24-h molekylære svingninger1,3,4 . Omkring 150 neuroner indeholdende disse molekylære ure form flyver et kredsløb til kontrollen døgnrytmen opførsel af voksen5. En meget enklere, men fuldt funktionelle døgnrytmen kredsløb bestående af 3 grupper af ur neuroner - 5 ventrale Lateral neuroner (LNvs; 4 PDF-positive LNvs og én PDF-negative LNv, se nedenfor), 2 dorsale Neuron 1s (DN1s) og 2 dorsale Neuron 2s (DN2s) - er til stede i den larve hjernen6,7.
Den simpel larve døgnrytmen kredsløb tilbyder en fremragende model til at studere samspillet mellem Inter neuronal kommunikation og den molekylære urværk. Ved hjælp af vores nyudviklede fluorescerende journalist PER-TDT, som efterligner niveauer af PER protein og dens subcellulært placering, vi har søgt at beskrive dynamikken i den molekylære urværk i forskellige ur neuron undergrupper i larve døgnrytmen kredsløb 8. desuden at vide nøglerolle neuropeptid pigment-spreder faktor (PDF) produceret af 4 LNvs i reguleringen af døgnrytmen på neuronal niveau9,10,11, vi ønskede at undersøge direkte effekt af PDF på de molekylære ure. Med henblik herpå udviklede vi en metode til at overvåge døgnrytmen genekspression rytmer i larve hjernen explant over flere dage af time-lapse Konfokal mikroskopi. Protokollen var også tilpasset farmakologiske assays til at teste effekten af PDF eller andre forbindelser på niveauet af PER-TDT. Således består tilpasning af at gøre hjernen eksplantat kultur tilgængeligt for drug application, øge tidsmæssige opløsning og imaging for en kortere varighed.
Ex vivo kultur af Drosophila hjerner af forskellige udviklingstrin har været tidligere etablerede12,13,14,15,16,17 ,18. Der henviser til, at disse protokoller er blevet anvendt for imaging forskellige biologiske fænomener, nogle af dem er ikke kompatible med billedbehandling med enkelt-celle opløsning eller understøtter ikke kulturen i mere end adskillige timer. Alternative metoder til at udføre langsigtede live imaging af døgnrytmen neuroner i Drosophila omfatter bioluminescens billeddannelse af Molekylær rytme19,20,21 og fluorescens billeddannelse af calcium indikator med lys ark mikroskopi22,23. Selvom bioluminescens imaging kan opnå højere tidsmæssige opløsning og lys ark mikroskopi kan være fleksibel for i vivo billeddannelse, de er begrænsede på den rumlige opløsning og kræver specialiseret mikroskop systemer.
Metoden beskrevet her er skræddersyet til at visualisere fluorescerende signaler i hele hjernen kultur med enkelt-celle opløsning over flere dage. Denne letkøbt og alsidig metode kunne tilpasses billedet kulturperler voksne flyve hjerner og for farmakologiske forsøg at studere mange forskellige problemer i Drosophila neurobiologi.

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	<tbody>
		<tr>
			<td>KH2PO4</td>
			<td>Sigma-Aldrich</td>
			<td>P5655</td>
			<td>I am not sure they are exactly the same ones we have in the lab. I chose &#34;suitable for insect cell culture&#34; whenever available</td>
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			<td>CaCl2</td>
			<td>Sigma-Aldrich</td>
			<td>C7902</td>
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			<td>MgSO4.7H2O</td>
			<td>Sigma-Aldrich</td>
			<td>230391</td>
			<td></td>
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		<tr>
			<td>NaCl</td>
			<td>Sigma-Aldrich</td>
			<td>S5886</td>
			<td></td>
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		<tr>
			<td>NaHCO3</td>
			<td>Sigma-Aldrich</td>
			<td>S5761</td>
			<td></td>
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		<tr>
			<td>D-(+) Glucose</td>
			<td>Sigma-Aldrich</td>
			<td>G7021</td>
			<td></td>
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			<td>Yeast extract</td>
			<td>Sigma-Aldrich</td>
			<td>Y1000</td>
			<td></td>
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			<td>Insulin</td>
			<td>Sigma-Aldrich</td>
			<td>I0516-5ML</td>
			<td></td>
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			<td>Penicillin-Streptomycin</td>
			<td>Sigma-Aldrich</td>
			<td>P4333</td>
			<td></td>
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		<tr>
			<td>BIS-TRIS</td>
			<td>Sigma-Aldrich</td>
			<td>B4429</td>
			<td></td>
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			<td>L-(&#8722;)-Malic acid</td>
			<td>Sigma-Aldrich</td>
			<td>M7397</td>
			<td></td>
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		<tr>
			<td>D-(+)-Trehalose dihydrate</td>
			<td>Sigma-Aldrich</td>
			<td>T0167</td>
			<td></td>
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		<tr>
			<td>Succinic acid</td>
			<td>Sigma-Aldrich</td>
			<td>S9512</td>
			<td></td>
		</tr>
		<tr>
			<td>Fumaric acid</td>
			<td>Sigma-Aldrich</td>
			<td>F8509</td>
			<td></td>
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		<tr>
			<td>&#945;-Ketoglutaric acid</td>
			<td>Sigma-Aldrich</td>
			<td>K1128</td>
			<td></td>
		</tr>
		<tr>
			<td>Non-heat-inactivated, Foetal Calf Serum (FCS) Mycoplasma and Virus screened</td>
			<td>BioConcept Ltd. Amimed</td>
			<td>2-01F30-I</td>
			<td></td>
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		<tr>
			<td>HEPES-KOH, pH 7.4</td>
			<td>E&#38;K Scientific Products</td>
			<td>EK-654011</td>
			<td></td>
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		<tr>
			<td>KCl</td>
			<td>Sigma-Aldrich</td>
			<td>P5405</td>
			<td></td>
		</tr>
		<tr>
			<td>NaH2PO4</td>
			<td>Sigma-Aldrich</td>
			<td>S5011</td>
			<td></td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Sigma-Aldrich</td>
			<td>S7903</td>
			<td></td>
		</tr>
		<tr>
			<td>Fibrinogen from bovine plasma</td>
			<td>Calbiochem (Merck)</td>
			<td>341573-1GM</td>
			<td>CAUTION: Harmful by inhalation, in contact with skin and if swallowed. Manipulate under laminar flow</td>
		</tr>
		<tr>
			<td>Thrombin from bovine plasma</td>
			<td>Sigma-Aldrich</td>
			<td>T9549</td>
			<td>CAUTION: Health Hazard, use gloves</td>
		</tr>
		<tr>
			<td>PDF, NH2-NSELINSLLSLPKNMNDA-OH</td>
			<td>Chi Scientific</td>
			<td>custom made</td>
			<td></td>
		</tr>
		<tr>
			<td>Vaccum grease</td>
			<td>Sigma-Aldrich</td>
			<td>18405</td>
			<td></td>
		</tr>
		<tr>
			<td>35 mm Dish, No. 1.5 Coverslip, 20 mm Glass Diameter, Uncoated</td>
			<td>MatTek</td>
			<td>P35G-1.5-20-C</td>
			<td></td>
		</tr>
		<tr>
			<td>Corning Falcon Easy-Grip Tissue Culture Dishes</td>
			<td>fisherscientific</td>
			<td>08-772A</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile 500 mL Steritop-GP 33 mm threaded bottle top filter, 0.22 &#956;m</td>
			<td>Millipore</td>
			<td>SCGPS05RE</td>
			<td></td>
		</tr>
		<tr>
			<td>Polytetrafluoroethylene (PTFE) film</td>
			<td>Dupont</td>
			<td>200A Teflon FEP Film</td>
			<td></td>
		</tr>
		<tr>
			<td>Millex-HV Syringe Filter Unit, 0.45&#160;&#181;m, PVDF, 33&#160;mm, gamma sterilized</td>
			<td>Millipore</td>
			<td>SLHV033RS</td>
			<td></td>
		</tr>
		<tr>
			<td>Millex-GV Syringe Filter Unit, 0.22&#160;&#181;m, PVDF, 33&#160;mm, gamma sterilized</td>
			<td>Millipore</td>
			<td>SLGV033RS</td>
			<td></td>
		</tr>
		<tr>
			<td>Three-well glass dissection dish</td>
			<td>Any company</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Fine forceps, size 5, Dumont</td>
			<td>Fine Science Tools</td>
			<td>11254-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Tandem scanner inverted TCS SP5 confocal microscope, with resonant scanner and HyD photo-multiplier detectors</td>
			<td>Leica microsystem CMS GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Temperature control chamber</td>
			<td>Life Imaging Services</td>
			<td>The CUBE &#38; BOX temperature control system, custom designed</td>
			<td></td>
		</tr>
		<tr>
			<td>Stage-top humidity controller</td>
			<td>Life Imaging Services</td>
			<td>custom made</td>
			<td></td>
		</tr>
		<tr>
			<td>Water Immersion Micro Dispenser: dispenser, extended micro-pump MP6 series and Autoimmersion Objective Controller software</td>
			<td>Leica microsystem CMS GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>SUM-stack creation and 3D correction drift plugin</td>
			<td>ImageJ software</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>10x iterative deconvolution</td>
			<td>AutoQuant and Imaris software</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

single-cell resolution fluorescence live imaging of drosophila circadian clocks in larval brain culture

Døgnrytmen pacemaker kredsløb orchestrates rytmiske adfærdsmæssige og fysiologiske udgange koordineres med miljømæssige stikord, som dag/nat cyklusser. Den molekylære ur inden for hver pacemaker neuron genererer døgnrytmen i genekspression, der ligger til grund for de rytmiske neuronal funktioner afgørende for driften af banen. Undersøgelse af de individuelle molekylære oscillatorer i forskellige underklasser af pacemaker neuroner og deres interaktion med neuronal signalering egenskaber giver en bedre forståelse af døgnrytmen pacemaker kredsløb. Her præsenterer vi en time-lapse fluorescerende mikroskopi tilgang udviklet til at overvåge den molekylære urværk i ur neuroner i kulturperler Drosophila larve hjernen. Denne metode giver mulighed for multi-dages optagelse af rytmer af genetisk kodet fluorescerende døgnrytmen reportere på én celle opløsning. Denne opsætning kan kombineres med farmakologiske manipulationer nøje analysere real-time svar af den molekylære ur til forskellige forbindelser. Ud over døgnrytmen giver denne multipurpose metode i kombination med kraftfulde Drosophila genetiske teknikker mulighed for at studere forskellige neuronal eller molekylære processer i levende hjernevæv.

Her, viser vi de repræsentative resultater af den langsigtede optagelse af døgnrytmen fluorescerende reporter i ex vivo larve hjernen kultur og live imaging resultaterne af PDF bad ansøgning på reporter udtryk.
Non-vandrer L3 larver at udtrykke den molekylære ur journalist PER-TDT og UAS-mCD8::YFP drevet af en ur neuron driver Clk (856)-gal4 (figur 1 c) var indblandede i LD. B...

Watch this Scientific Journal Video about Single-cell Resolution Fluorescence Live Imaging of Drosophila Circadian Clocks in Larval Brain Culture at JoVE.com

Single-cell Resolution Fluorescence Live Imaging of Drosophila Circadian Clocks in Larval Brain Culture

Målet med denne protokol er at etablere ex vivo Drosophila larve hjernen kultur optimeret til at overvåge døgnrytmen Molekylær rytme med langsigtede fluorescens time-lapse billeddannelse. Anvendelsen af denne metode til farmakologisk assays behandles også.

Encellede opløsning fluorescens Live Imaging af Drosophila døgnrytmen ure i larve hjernen kultur

The circadian pacemaker circuit orchestrates rhythmic behavioral and physiological outputs coordinated with environmental cues, such as day/night cycles. ...

Research

JoVE Journal

Neuroscience

Single-cell Resolution Fluorescence Live Imaging of Drosophila Circadian Clocks in Larval Brain Culture

Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations

Visualization of Larval Segmental Nerves in 3rd Instar Drosophila Larval Preparations

Drosophila melanogaster is emerging as a powerful model system for studying the development and function of the nervous system, particularly because of ...

Live-cell Imaging of Sensory Organ Precursor Cells in Intact Drosophila Pupae

Live-cell Imaging of Sensory Organ Precursor Cells in Intact Drosophila Pupae

Since the discovery of Green Fluorescent Protein (GFP), there has been a revolutionary change in the use of live-cell imaging as a tool for understanding ...

A Rapid Approach to High-Resolution Fluorescence Imaging in Semi-Thick Brain Slices

A fundamental goal to both basic and clinical neuroscience is to better understand the identities, molecular makeup, and patterns of connectivity that are ...

Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics

Morphological Analysis of Drosophila Larval Peripheral Sensory Neuron Dendrites and Axons Using Genetic Mosaics

Morfologiske Analyse af Drosophila Larver Perifer sensorisk Neuron dendritter og axoner anvende genetiske Mosaikker

Nervous system development requires the correct specification of neuron position and identity, followed by accurate neuron class-specific dendritic ...

Non-Laser Capture Microscopy Approach for the Microdissection of Discrete Mouse Brain Regions for Total RNA Isolation and Downstream Next-Generation Sequencing and Gene Expression Profiling

Non-Laser Capture Microscopy Approach for the Microdissection of Discrete Mouse Brain Regions for Total RNA Isolation and Downstream Next-Generation Sequencing and Gene Expression Profiling  |

Ikke-Laser Capture Mikroskopi Tilgang til mikrodissektion af diskrete Mouse Brain Regioner for Total RNA Isolation og downstream næste generation sekventering og Gene Expression Profiling

As technological platforms, approaches such as next-generation sequencing, microarray, and qRT-PCR have great promise for expanding our understanding of ...

High-resolution Live Imaging of Cell Behavior in the Developing Neuroepithelium

Høj opløsning Live-Imaging af celle adfærd i udviklingslandene neuroepitel

The embryonic spinal cord consists of cycling neural progenitor cells that give rise to a large percentage of the neuronal and glial cells of the central ...

Ex vivo Live Imaging of Single Cell Divisions in Mouse Neuroepithelium

Ex vivo Live Imaging of Single Cell Divisions in Mouse Neuroepithelium

Ex vivo Live-Imaging af Single celledelinger i Mouse neuroepithelium

We developed a system that integrates live imaging of fluorescent markers and culturing slices of embryonic mouse neuroepithelium. We took advantage of ...

Live Imaging of Mitosis in the Developing Mouse Embryonic Cortex

Live-Imaging mitosens i udviklingslandene mus embryonale Cortex

Although of short duration, mitosis is a complex and dynamic multi-step process fundamental for development of organs including the brain. In the ...

Super-resolution Imaging of Neuronal Dense-core Vesicles

Super-opløsning Imaging af neuronale Dense-core blærer

Detection of fluorescence provides the foundation for many widely utilized and rapidly advancing microscopy techniques employed in modern biological and ...

Live Imaging of Drosophila Larval Neuroblasts

Live Imaging of Drosophila Larval Neuroblasts

Live-Imaging af<em&gt; Drosophila</em&gt; Larvetilstand neuroblasts

Stem cells divide asymmetrically to generate two progeny cells with unequal fate potential: a self-renewing stem cell and a differentiating cell. Given ...