Name: culture of adult transgenic zebrafish retinal explants for live-cell imaging by multiphoton microscopy
Uploaded: 2017-02-24T15:00:00
Description: Watch this Scientific Journal Video about Culture of Adult Transgenic Zebrafish Retinal Explants for Live-cell Imaging by Multiphoton Microscopy at JoVE.com

Vi setter pris på støtten fra William Archer og Notre Dame Integrated Imaging Facility. Spesiell takk rettes til Freimann Life Sciences teknikere for deres kontinuerlig hjelp og deres omsorg og dyrehold av sebrafisk. Denne studien ble støttet med tilskudd fra National Eye Institute of NIH til DRH (R01-EY018417, R01-EY024519) og Senter for Sebrafisk Research, University of Notre Dame, Notre Dame, IN.

Merk: Sebrafisk ble reist og vedlikeholdes i Notre Dame Sebrafisk-anlegget i Freimann Life Sciences Center. Metodene som er beskrevet i dette manuskriptet er godkjent av University of Notre Dame Animal Care og bruk Committee og er i samsvar med uttalelsen for bruk av dyr i et syn forskning av Association for Research in Vision og oftalmologi. 1. Solutions <ol><li> Forbered 70% etanol for å sterilisere vevskultur panseret og noe utstyr / reagenser som overføres inn i vev kultur hette. </l...

<ol>
	<li>Fausett, B. V., Goldman, 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+role+for+alpha1+tubulin-expressing+Muller+glia+in+regeneration+of+the+injured+zebrafish+retina.">A role for alpha1 tubulin-expressing Muller glia in regeneration of the injured zebrafish retina.</a> J.Neurosci. 26 (23), 6303-6313 (2006).</li><li>Kassen, S. 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=Time+course+analysis+of+gene+expression+during+light-induced+photoreceptor+cell+death+and+regeneration+in+albino+zebrafish.">Time course analysis of gene expression during light-induced photoreceptor cell death and regeneration in albino zebrafish.</a> Dev.Neurobiol. 67 (8), 1009-1031 (2007).</li><li>Vihtelic, T. S., Hyde, D. 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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=Multiphoton+imaging+of+chick+retinal+development+in+relation+to+gap+junctional+communication.">Multiphoton imaging of chick retinal development in relation to gap junctional communication.</a> J.Physiol. 585 (Pt 3), 711-719 (2007).</li><li>Surzenko, N., Crowl, T., Bachleda, A., Langer, L., Pevny, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SOX2+maintains+the+quiescent+progenitor+cell+state+of+postnatal+retinal+Muller+glia.">SOX2 maintains the quiescent progenitor cell state of postnatal retinal Muller glia.</a> Development. 140 (7), 1445-1456 (2013).</li><li>Nickerson, P. E., 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=Live+imaging+and+analysis+of+postnatal+mouse+retinal+development.">Live imaging and analysis of postnatal mouse retinal development.</a> BMC Dev.Biol. 13, 24 (2013).</li><li>Suzuki, S. C., Bleckert, A., Williams, P. R., Takechi, M., Kawamura, S., Wong, R. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cone+photoreceptor+types+in+zebrafish+are+generated+by+symmetric+terminal+divisions+of+dedicated+precursors.">Cone photoreceptor types in zebrafish are generated by symmetric terminal divisions of dedicated precursors.</a> Proc.Natl.Acad.Sci.U.S.A. 110 (37), 15109-15114 (2013).</li><li>Johnson, T. V., Martin, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+characterization+of+an+adult+retinal+explant+organotypic+tissue+culture+system+as+an+in+vitro+intraocular+stem+cell+transplantation+model.">Development and characterization of an adult retinal explant organotypic tissue culture system as an in vitro intraocular stem cell transplantation model.</a> Invest.Ophthalmol.Vis.Sci. 49 (8), 3503-3512 (2008).</li><li>Bull, N. D., Johnson, T. V., Welsapar, G., DeKorver, N. W., Tomarev, S. I., Martin, K. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+an+adult+rat+retinal+explant+model+for+screening+of+potential+retinal+ganglion+cell+neuroprotective+therapies.">Use of an adult rat retinal explant model for screening of potential retinal ganglion cell neuroprotective therapies.</a> Invest.Ophthalmol.Vis.Sci. 52 (6), 3309-3320 (2011).</li><li>Kustermann, S., Schmid, S., Biehlmaier, O., Kohler, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Survival,+excitability,+and+transfection+of+retinal+neurons+in+an+organotypic+culture+of+mature+zebrafish+retina.">Survival, excitability, and transfection of retinal neurons in an organotypic culture of mature zebrafish retina.</a> Cell Tissue Res. 332 (2), 195-209 (2008).</li><li>Williams, P. R., Morgan, J. L., Kerschensteiner, D., Wong, R. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+imaging+of+retinal+whole+mounts.">In vitro imaging of retinal whole mounts.</a> Cold Spring Harb Protoc. 2013, (2013).</li><li>Johnson, T. V., Oglesby, E. N., Steinhart, M. R., Cone-Kimball, E., Jefferys, J., Quigley, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time-Lapse+Retinal+Ganglion+Cell+Dendritic+Field+Degeneration+Imaged+in+Organotypic+Retinal+Explant+Culture.">Time-Lapse Retinal Ganglion Cell Dendritic Field Degeneration Imaged in Organotypic Retinal Explant Culture.</a> Invest.Ophthalmol.Vis.Sci. 57 (1), 253-264 (2016).</li><li>Roehlecke, C., Schumann, U., Ader, M., Knels, L., Funk, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+blue+light+on+photoreceptors+in+a+live+retinal+explant+system.">Influence of blue light on photoreceptors in a live retinal explant system.</a> Mol.Vis. 17, 876-884 (2011).</li><li>Coutu, D. L., Schroeder, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Probing+cellular+processes+by+long-term+live+imaging--historic+problems+and+current+solutions.">Probing cellular processes by long-term live imaging--historic problems and current solutions.</a> J.Cell.Sci. 126 (Pt 17), 3805-3815 (2013).</li><li>Thomas, J. L., Thummel, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+light+damage+paradigm+for+use+in+retinal+regeneration+studies+in+adult+zebrafish.">A novel light damage paradigm for use in retinal regeneration studies in adult zebrafish.</a> J.Vis.Exp. 80, e51017 (2013).</li><li>Conner, C., Ackerman, K. M., Lahne, M., Hobgood, J. S., Hyde, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repressing+notch+signaling+and+expressing+TNFalpha+are+sufficient+to+mimic+retinal+regeneration+by+inducing+Muller+glial+proliferation+to+generate+committed+progenitor+cells.">Repressing notch signaling and expressing TNFalpha are sufficient to mimic retinal regeneration by inducing Muller glial proliferation to generate committed progenitor cells.</a> J.Neurosci. 34 (43), 14403-14419 (2014).</li><li>Nelson, C. M., Gorsuch, R. A., Bailey, T. J., Ackerman, K. M., Kassen, S. C., Hyde, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stat3+defines+three+populations+of+Muller+glia+and+is+required+for+initiating+maximal+muller+glia+proliferation+in+the+regenerating+zebrafish+retina.">Stat3 defines three populations of Muller glia and is required for initiating maximal muller glia proliferation in the regenerating zebrafish retina.</a> J.Comp.Neurol. 520 (18), 4294-4311 (2012).</li><li>Gorsuch, R. A., Hyde, D. R. <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+Muller+glial+dependent+neuronal+regeneration+in+the+damaged+adult+zebrafish+retina.">Regulation of Muller glial dependent neuronal regeneration in the damaged adult zebrafish retina.</a> Exp.Eye Res. 123, 131-140 (2014).</li><li>Zhao, X. F., Wan, J., Powell, C., Ramachandran, R., Myers, M. G., Goldman, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Leptin+and+IL-6+family+cytokines+synergize+to+stimulate+Muller+glia+reprogramming+and+retina+regeneration.">Leptin and IL-6 family cytokines synergize to stimulate Muller glia reprogramming and retina regeneration.</a> Cell.Rep. 9 (1), 272-284 (2014).</li><li>Wan, J., Ramachandran, R., Goldman, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HB-EGF+is+necessary+and+sufficient+for+Muller+glia+dedifferentiation+and+retina+regeneration.">HB-EGF is necessary and sufficient for Muller glia dedifferentiation and retina regeneration.</a> Dev.Cell. 22 (2), 334-347 (2012).</li><li>Lenkowski, J. 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Studier som undersøker de mekanismene som styrer regenerering av skadet voksen sebrafisk netthinnen hovedsakelig brukte immunocytokjemiske metoder 5, 25, 26, 27, 28, 29, 30. Etablering forhold til kultur retinal explants og å opptre live-cell imaging på fenomener som INM, gi oss en tekni...

I motsetning til mennesker, sebrafisk (Danio rerio) viser en robust regenerering svar på celledød av netthinnens nerveceller 1, 2, 3, 4. Tumornekrosefaktor α, et signalmolekyl som frigjøres fra å dø netthinnens nerveceller induserer Müller gliaceller bosatt i basal Indre Nuclear Layer (INL) av netthinnen, for å spre fem og produsere nevrale stamceller som fortsetter å spre seg før differensiere inn i nevronale celle typer som døde 2, 3, 4. Under den proliferative fase av regenereringen respons, kjerner av Müller gliaceller og deres avledet neuronal forløperceller gjennomgå et repeterende mønster trekkende i fase med cellesyklusen (Interkinetic Nuclear Migration, INM) 6 </sup<html> &gt; 7. Kjerner plassert i basal INL kopiere deres DNA før migrere til ytre Nuclear Layer (onl) hvor de deler før de oppstår atomkjerner tilbake basalt til INL. Denne fremgangsmåten ble først beskrevet under neuroepithelial utvikling ved hjelp av histologiske metoder, mens levende celle bildedannende fremgangsmåter senere bekreftet tolkningen av Sauer 8, 9, 10, 11, 12. Både histokjemiske og lever-cellebilde tilnærminger er blitt benyttet for å bestemme mekanismene bak INM og dens funksjon i å utvikle neuroepithelia herunder netthinnen 9, 11, 12, 13. Imidlertid har de mekanismene som styrer INM i voksen regenererende hinnen ikke blitt undersøkt i mye detaljxref &quot;&gt; 6, 7. Lev-cell imaging vil være en uvurderlig måte å fremme vår kunnskap om signalveier som styrer INM i voksen regenererende hinnen. Inntil nylig var levende celle avbildning av INM i netthinnen begrenset til enten levende sebrafisk embryo eller foster dama eller postnatal mus retinal explants 9, 10, 11, 12, 14, 15, 16. Mens retinal explants fra voksne dyr av ulike arter, inkludert mus, rotte og sebrafisk har blitt utnyttet for annen celle biologiske tilnærminger 17, 18, 19, 20, live-cell imaging eksperimenter med retinal eksplanter hahar vært begrenset til korte perioder av gangen, og har ikke blitt utført kontinuerlig over flere timer 21, 22. Her beskriver vi en detaljert protokoll til kultur lys-skadet voksne sebrafisk netthinner å opptre live-celle bildebehandling eksperimenter overvåking INM ved hjelp av multi-foton mikroskopi 6. Levende celler bilde tilnærminger er fordelaktig i forhold immunhistokjemiske metoder når undersøke hvilke mekanismer som styrer INM, som dynamikken i INM, f.eks hastigheter kan bli påvirket snarere enn plasseringen av mitose, som ville potensielt ikke oppdages ved hjelp immunocytochemistry. I fremtiden, har denne fremgangsmåte også potensiale til å bli modifisert for å studere andre dynamiske prosesser i løpet av retinal regenerasjon, slik som fagocytose av døende fotoreseptorene hos Müller gliaceller eller oppførselen til mikroglia.

<table border="0" cellpadding="0" cellspacing="0" width="876">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:491px;">Dumont forceps size #5</td>
			<td style="width:201px;">World precision instruments</td>
			<td align="right" style="width:113px;">14098</td>
			<td style="width:72px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dumont forceps size #5, 45&#176; angle</td>
			<td>World precision instruments</td>
			<td align="right">14101</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">McPherson-Vannas scissors</td>
			<td>World precision instruments</td>
			<td align="right">501233</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Fluordishes</td>
			<td>World precision instruments</td>
			<td>FD35-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stereomicroscope</td>
			<td>Nikon&#160;</td>
			<td>SMZ-1B</td>
			<td>similar type of dissection stereomicroscope will work</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Biological Safety Cabinet class type A2</td>
			<td>Labconco</td>
			<td></td>
			<td>equivalent type will work</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">tissue culture incubator</td>
			<td>Thermoscientific</td>
			<td>HEPA-class 100</td>
			<td>equivalent type will work</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sylvania fluorescent lamps OSFP5835HOECO</td>
			<td>Bulbtronics</td>
			<td align="right">31850</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.2 &#181;m pore-size Acrodisc syringe filter</td>
			<td>VWR</td>
			<td align="right">4192</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">10 ml Luer-lok syringe</td>
			<td>VWR</td>
			<td>BD309604</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">60 ml Luer-lok syringe</td>
			<td>VWR</td>
			<td>BD309653</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">NaHCO3</td>
			<td>FischerScientific</td>
			<td>S233-500</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">CaCl2</td>
			<td>ThermoScientific</td>
			<td>C79-500</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">MgCl2</td>
			<td>EMD Millipore</td>
			<td align="right">5980</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;">HBSS w/o Ca2+/Mg2+, w/o phenol red, Gibco</td>
			<td>ThermoScientific</td>
			<td>14175-095</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MEM w/o phenol red, Gibco</td>
			<td>ThermoScientific</td>
			<td>5100-038</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Horse serum, heat-inactivated</td>
			<td>ThermoScientific</td>
			<td>26050-070</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">penicillin/streptomycin</td>
			<td>VWR</td>
			<td>16777-164</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ultrapure low melting point agarose</td>
			<td>ThermoScientific</td>
			<td>16520-100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ethanol, absolute</td>
			<td>ThermoScientific</td>
			<td>BP2818-4</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-phenoxyethanol</td>
			<td>Sigma</td>
			<td align="right">77699</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Corning Cell-Tak cell and tissue adhesive&#160;</td>
			<td>VWR</td>
			<td align="right">354240</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">refractive index liquid&#160;</td>
			<td>Cargille Lab</td>
			<td>1803Y</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nikon A1 multiphoton microscope equipped with a MaiTai infrared laser</td>
			<td>Nikon</td>
			<td></td>
			<td>equivalent system will work</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">40x Apo long-distance water immersion objective (N.A. 1.15)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">environmental chamber equipped with insert for 35 mm petridishes</td>
			<td>Okolab</td>
			<td></td>
			<td>equivalent system will work</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NIS analysis software</td>
			<td>Nikon</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

culture of adult transgenic zebrafish retinal explants for live-cell imaging by multiphoton microscopy

En endogen regenerering Programmet er initiert av Müller gliaceller i den voksne sebrafisk (Danio rerio) hinnen etter neuronal skade og død. De Müller gliaceller re-gå inn i cellesyklus og produsere nevrale stamceller som gjennomgår påfølgende runder av celledelinger og differensiere inn de tapte nevronale celletyper. Både Müller gliaceller og nevronale stamfar cellekjerner kopiere deres DNA og gjennomgå mitose i forskjellige steder i netthinnen, dvs. de vandrer mellom basal Indre Nuclear Layer (INL) og Ytre Nuclear Layer (onl), henholdsvis i en prosess beskrevet som Interkinetic Nuclear Migration (INM). INM har hovedsakelig blitt studert i utviklingshinnen. For å undersøke dynamikken i INM i den voksne regenererende sebrafisk netthinnen i detalj, er levende celle avbildning av fluoresceinmerkede Müller gliaceller / nevrale stamceller nødvendig. Her gir vi betingelsene for å isolere og kultur rygg netthinnefra Tg [GFAP: nGFP] mi2004 sebrafisk som ble utsatt for konstant intens lys for 35 timer. Vi viser også at disse retina kulturer er levedyktig for å utføre levende celle bildebehandling eksperimenter, kontinuerlig å anskaffe z-stack-bilder i hele tykkelsen av retinal eksplantat i opptil 8 timer under anvendelse av multiphoton mikroskopi for å overvåke vandringsadferd av GFAP: nGFP-positive celler . I tillegg beskriver vi detaljene til å utføre post-bildeanalyse for å bestemme hastigheten av apikale og basal INM. For å oppsummere, etablerte vi forutsetninger for å studere dynamikken i INM i en voksen modell av neuronal regenerering. Dette vil fremme vår forståelse av denne viktige cellulære prosessen og tillate oss å bestemme hvilke mekanismer som styrer INM.

Isoleringen av netthinnen i henhold til fremgangsmåten beskrevet i skjematisk på figur 1 tillater dyrkning av en flat rygg retina fra lys-skadet voksen Tg [GFAP: nGFP] mi2004 sebrafisk i løpet av en periode på minst 24 timer i en 5% CO 2 / luft. Disse flat-monterte retinal explants kan brukes til bildefokusplan på dype vev nivåer. Et eksempel er Müller gliaceller / nervestamcellekjerner merkes med GFP fra Müller glia-spesifikke prom...

Watch this Scientific Journal Video about Culture of Adult Transgenic Zebrafish Retinal Explants for Live-cell Imaging by Multiphoton Microscopy at JoVE.com

Culture of Adult Transgenic Zebrafish Retinal Explants for Live-cell Imaging by Multiphoton Microscopy

Sebrafisk retinal regenerasjon har for det meste blitt undersøkt ved hjelp av faste netthinner. Men dynamiske prosesser som interkinetic atom migrasjon oppstår under regenerativ respons og krever levende celle bildebehandling for å undersøke de underliggende mekanismene. Her beskriver vi kultur og bilde forhold til å overvåke Interkinetic Nuclear Migration (INM) i sanntid ved hjelp multiphoton mikroskopi.

Culture of Adult Transgene Sebrafisk Netthinne explants for Live-celle Imaging av multiphoton Mikros

An endogenous regeneration program is initiated by M&#252;ller glia in the adult zebrafish (Danio rerio) retina following neuronal damage and death. The ...

Research

JoVE Journal

Developmental Biology

Kidney Regeneration in Adult Zebrafish by Gentamicin Induced Injury

Nyre Regeneration i Adult Sebrafisk av Gentamicin Induced Injury

The kidney is essential for fluid homeostasis, blood pressure regulation and filtration of waste from the body. The fundamental unit of kidney function is ...

Culture of Embryonic Mouse Cochlear Explants and Gene Transfer by Electroporation

Culture of Embryonale Mus Cochlea eksplanter og Gene Transfer ved Electroporation

Auditory hair cells located within the mouse organ of Corti detect and transmit sound information to the central nervous system. The mechanosensory hair ...

Whole Retinal Explants from Chicken Embryos for Electroporation and Chemical Reagent Treatments

Hele Retinal eksplantater fra kylling embryoer for Electroporation og kjemisk reagens behandlinger

The retina is a good model for the developing central nervous system. The large size of the eye and most importantly the accessibility for experimental ...

Tracking Cells in GFP-transgenic Zebrafish Using the Photoconvertible PSmOrange System

Sporing celler i GFP-transgene Sebrafisk Bruke Photoconvertible PSmOrange System

The rapid development of transparent zebrafish embryos (Danio rerio) in combination with fluorescent labelings of cells and tissues allows visualizing ...

Isolation and Culture of Adult Zebrafish Brain-derived Neurospheres

Isolasjon og Culture of Adult Sebrafisk Brain-avledet Neurosfærer

The zebrafish is a highly relevant model organism for understanding the cellular and molecular mechanisms involved in neurogenesis and brain regeneration ...

Analysis of Zebrafish Kidney Development with Time-lapse Imaging Using a Dissecting Microscope Equipped for Optical Sectioning

Analyse av Sebrafisk Nyre Development med Time-lapse Imaging Ved hjelp av en dissekere mikroskop Utstyrt for Optical Seksjonering

In order to understand organogenesis, the spatial and temporal alterations that occur during development of tissues need to be recorded. The method ...

In Vivo Imaging of Transgenic Gene Expression in Individual Retinal Progenitors in Chimeric Zebrafish Embryos to Study Cell Nonautonomous Influences

In Vivo Imaging of Transgenic Gene Expression in Individual Retinal Progenitors in Chimeric Zebrafish Embryos to Study Cell Nonautonomous Influences

In vivo avbildning av transgene genuttrykk i Individuell Netthinnestamfedre i kimære Sebrafisk embryoer å studere Cell Nonautonomous Influences

The genetic and technical strengths have made the zebrafish vertebrate a key model organism in which the consequences of gene manipulations can be traced ...

Live Imaging of Mouse Secondary Palate Fusion

The fusion of the secondary palatal shelves to form the intact secondary palate is a key process in mammalian development and its disruption can lead to ...

Multimodal Hierarchical Imaging of Serial Sections for Finding Specific Cellular Targets within Large Volumes

Flere hierarkisk Imaging av føljetong delene for å finne konkrete mobilnettet mål i store volumer

Targeting specific cells at ultrastructural resolution within a mixed cell population or a tissue can be achieved by hierarchical imaging using a ...

Live Cell Imaging of Chromosome Segregation During Mitosis

Levende celle Imaging av kromosom segregering under mitose

Chromosomes must be reliably and uniformly segregated into daughter cells during mitotic cell division. Fidelity of chromosomal segregation is controlled ...