Name: modified roller tube method for precisely localized and repetitive intermittent imaging during long-term culture of brain slices in an enclosed system
Uploaded: 2017-12-28T13:00:00
Description: Watch this Scientific Journal Video about Modified Roller Tube Method for Precisely Localized and Repetitive Intermittent Imaging During Long-term Culture of Brain Slices in an Enclosed System at JoVE

Dyr bruk følger godkjent avl og dyr bruk protokoller som svarer til Animal Care og bruk retningslinjer av Colorado State University.
Merk: Protokollen nedenfor beskriver metoden forberedelse og kultur for langsiktig inkubasjon og intermitterende avbilding av hippocampus skiver. En enkelt hippocampus skive er knyttet til et spesielt forberedt photoetched dekkglassvæske bruker en plasma blodpropp, og deretter coverslips er forseglet på den flate siden av en boret ut valse røret, som oppretth...

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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+integration+of+rAAV+vector+into+the+AAVS1+region.">Targeted integration of rAAV vector into the AAVS1 region.</a> Virology. 433 (2), 356-366 (2012).</li><li>Akerboom, 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=Optimization+of+a+GCaMP+calcium+indicator+for+neural+activity+imaging.">Optimization of a GCaMP calcium indicator for neural activity imaging.</a> J Neurosci. 32 (40), 13819-13840 (2012).</li><li>Kim, J. 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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formation+of+actin-ADF/cofilin+rods+transiently+retards+decline+of+mitochondrial+potential+and+ATP+in+stressed+neurons.">Formation of actin-ADF/cofilin rods transiently retards decline of mitochondrial potential and ATP in stressed neurons.</a> Am J Physiol Cell Physiol. 291 (5), 828-839 (2006).</li><li>Cichon, 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=Cofilin+aggregation+blocks+intracellular+trafficking+and+induces+synaptic+loss+in+hippocampal+neurons.">Cofilin aggregation blocks intracellular trafficking and induces synaptic loss in hippocampal neurons.</a> J Biol Chem. 287 (6), 3929-3939 (2012).</li><li>Stine, W. B., Dahlgren, K. N., Krafft, G. A., LaDu, M. J. <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+characterization+of+conditions+for+amyloid-beta+peptide+oligomerization+and+fibrillogenesis.">In vitro characterization of conditions for amyloid-beta peptide oligomerization and fibrillogenesis.</a> J Biol Chem. 278 (13), 11612-11622 (2003).</li><li>Maloney, M. T., Minamide, L. S., Kinley, A. W., Boyle, J. A., Bamburg, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beta-secretase-cleaved+amyloid+precursor+protein+accumulates+at+actin+inclusions+induced+in+neurons+by+stress+or+amyloid+beta:+a+feedforward+mechanisms+for+Alzheimer's+disease.">Beta-secretase-cleaved amyloid precursor protein accumulates at actin inclusions induced in neurons by stress or amyloid beta: a feedforward mechanisms for Alzheimer's disease.</a> J Neurosci. 25 (49), 11313-11321 (2005).</li><li>Davis, R. 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Valse røret metoden beskrevet her gir langsiktig dyrking og høy oppløsning live bildebehandling av skiver hjernevev. En viktig sak med SKIVE teknikken som brukes her er i montering og vedlikehold av skiver. Dekkglassvæske belegg som støtter skive vedheft, fremme skive tynning ved å forbedre resultatet av neurites og migrering av celler av stykket; dermed unngikk vi bruk av disse underlag. Innsetting av amino grupper på glasset av behandling med 3-aminopropyltriethoxysilane bedre etterlevelse av skiver, men for lit...

Primære kultur dissosiert neurons fra regioner av gnager hjernen er et viktig verktøy brukes av forskere til å observere Svar å pathologically innblandet stimuli. Men har slike studier ulempen ved å se på nerveceller i bare 2D og uten deres glial støttesystem. Videre med mindre vokst under forhold med svært høy tetthet (640 neurons/mm2 eller ca 16% av arealet) der det blir umulig å følge tilfeldig resultatet av en dendrite eller axon for mer enn en kort avstand fra celle kroppen, hippocampus neuronal levedyktighet synker over 4 uker betydelig1, begrenser bruken av dissosiert kulturer for utvidede studier av alder-relaterte sykdommer. Dyrking av skiver forberedt fra gnager hjernen er et attraktivt alternativ som overvinner disse begrensningene ved å vedlikeholde en organisert celle arkitektur og levedyktigheten for uker eller måneder. For å opprettholde mange regioner av gnager hjernen i skive kultur har vært beskrevet2.
To store metoder brukes for langsiktig kultur av hjernen skiver: dyrking på membraner på luft-flytende grensesnitt3 eller dyrking på coverslips i forseglet rør kan rotere en valse inkubator å gi lufting4. Skiver kultivert på membraner kan avbildes direkte med høy oppløsning fluorescens mikroskopi bruke oppreist mikroskop og vann nedsenking mål5. Alternativt har skiver kultivert på membraner blitt overført til glass bunn retter å oppnå god oppløsning av dendrittiske spines bruker en invertert mikroskop6. Begge metodene Imaging skiver dyrket på membraner er imidlertid åpne systemer som krever middels endringer og bruker ofte soppdrepende og/eller antibiotika for å forhindre eller redusere forurensning5,6. Skiver på en membran i luften mellomstore grensesnittet opprettholde gode morfologi og overlevelse, men tilbake til nøyaktige under repeterende bildebehandling på forstørring er svært vanskelig med mindre eksperimentet følger bare små grupper av celler uttrykke merketråd fluorescerende. Selv om skiver dyrket på membraner har blitt brukt med viral-mediert uttrykk effekter av transgener5,6, kan biosafety protokoller kreve en lukket kultur-systemet benyttes for visse viral vektorer som brukes for uttrykke fluorescently merket proteiner og journalister av cellen fysiologi. Videre krever nedsenking mål dekontaminering mellom eksempler som skal følges i kultur5. En stor anvendelse av membran grensesnitt kulturer er å kombinere høy oppløsning bildebehandling med elektrofysiologi på gang poeng7.
Metoden valse røret med coverslips inne i plast røret tillater ikke noen elektrofysiologi eller høy-resolution tenkelig uten å fjerne dekkglassvæske. Dermed er denne metoden oftest brukt til langsiktige studier der etter fiksering observasjoner er gjort8. Beskrevet her er en metode som utnytter valse røret kultur teknikken, men på boret ut rør med skiver på coverslips som kan avbildes gjentagelser så lenge kulturer opprettholdes. Lukkede systemet krever ingen medium endring for bildebehandling og utnytter photoetched coverslips for å gi fiducial merker at imaging på forstørring, etter dager eller uker, presis feltene tidligere fotografert.
Vi bruker denne metoden for å undersøke endringer gnager prefiks, en stor hjerne region involvert inne hukommelse og læring. Gnager hippocampus er ofte studert som en modell for patologisk eller alder-relaterte endringer observert under utviklingen av kognitiv svekkelse9, for eksempel de som oppstår i Annonsen. Vår metode er spesielt godt egnet til å studere patologiske forandringer som utvikler i et enkelt stykke over tid som respons til endringer i miljøet, slik som økning i Aβ-peptider som er karakteristisk for AD8. En patologi forbundet med menneskelige og gnager AD hjernen er tilstedeværelsen av cofilin-utgangen aggregater og stenger, sistnevnte som inneholder bunter av filamenter som cofilin og utgangen er i en 1:1 molar forholdet10,11, 12. stenger har blitt observert i fast skiver av rotte hippocampus etter Aβ behandling, samt i en levende gnagere hjerne skive uttrykke cofilin-GFP utsatt for hypoksi8, og de kan bidra til at synaptic dysfunksjon i AD og slag. Her bruker vi denne nye culturing metoden for å observere den gang forløp og distribusjon i skiver av uttrykt eksogene chimeric fluorescerende proteiner introdusert av forskjellige virus. Vi deretter bruke neuronal bestemte uttrykk for en cofilin reporter konstruere å følge utviklingen av cofilin stang og samlet patologi i hippocampus skiver i respons på behandling med løselig Aβ oligomers (Aβo).

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			<td height="40" style="height:40px;width:280px;">Bottoms from 15 cm culture dishes</td>
			<td style="width:220px;">VWR Scientific</td>
			<td style="width:191px;">25384-326</td>
			<td style="width:845px;"></td>
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			<td height="40" style="height:40px;">Phillips Head Machine Screws (#10-32)</td>
			<td>Ace Hardware</td>
			<td></td>
			<td>2.5&#34; long and 3/16&#34; in diameter</td>
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			<td height="40" style="height:40px;">Flat Washers #10</td>
			<td>ACE Hardware</td>
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			<td height="40" style="height:40px;">Machine Screw Nuts (#10-32)</td>
			<td>ACE Hardware</td>
			<td></td>
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			<td height="40" style="height:40px;">Rubber Grommets&#160;</td>
			<td>ACE Hardware</td>
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			<td>5/16&#34;, thick; 5/8&#34;, hole diameter; 1.125&#34;, OD</td>
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			<td height="40" style="height:40px;">Polyethylene tubing (5/16&#34;; OD, 3/16&#34;; ID)</td>
			<td>ACE Hardware</td>
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			<td>Cut to 1.8&#34; length</td>
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			<td>ACE Hardware</td>
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			<td>Ace Hardware</td>
			<td>ProTech Model 1201</td>
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			<td>VWR</td>
			<td>62407-076</td>
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			<td height="40" style="height:40px;">Drill bits, 3 mm, 6 mm and 15 mm&#160;</td>
			<td>Ace Hardware</td>
			<td>Diablo freud brand</td>
			<td>Drill bits for cutting plastic.</td>
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			<td>Ace Hardware</td>
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			<td>Ace Harware</td>
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			<td height="40" style="height:40px;">Spring clips, 16 mm snap holder</td>
			<td>Ace Hardware</td>
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			<td>Ace Hardware</td>
			<td>26307</td>
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		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:280px;">Adhesive Silicone Sheet (Secure Seal)</td>
			<td style="width:220px;">Grace Bio-Labs</td>
			<td style="width:191px;">666581</td>
			<td>0.5 mm Thickness</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">6 mm hole punch</td>
			<td>Office Max</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">12 mm hole punch</td>
			<td>thepunchbunch.com</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">70% Ethanol</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Phototeched Coverslips, 12 mm diameter</td>
			<td>Bellco Glass, Inc.</td>
			<td>1916-91012</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Bunsen Burner</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Absolute Ethanol</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Nanopure Water</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">3-aminopropyltriethoxylane</td>
			<td>Sigma-Aldrich</td>
			<td>A3648</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Acetone</td>
			<td>Sigma-Aldrich</td>
			<td>179124</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">#5 Dumont Forceps</td>
			<td>Fine Science Tools</td>
			<td>11251-30</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">McIlwain Tissue Chopper</td>
			<td>Ted Pella, Inc.</td>
			<td>10180</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Double Edge Razor Blades</td>
			<td>Ted Pella, Inc.</td>
			<td>121-6</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Whatman Filter Paper</td>
			<td>VWR</td>
			<td>28450-182</td>
			<td>Cut into 5.8 cm diameter circles</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Poly-chloro-trifluoro-ethylene (Aclar)</td>
			<td>Ted Pella, Inc.</td>
			<td>10501-10</td>
			<td>Cut into 5.8 cm diameter circles</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">#21 Surgical Blade</td>
			<td>VWR Scientific</td>
			<td>25860-144</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">#5 Dumont Forceps</td>
			<td>Fine Science Tools</td>
			<td>11251-30</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Spatula, stainless with tapered end</td>
			<td>VWR</td>
			<td>82027-518</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Gey&#39;s Balanced Salt Solution</td>
			<td>Sigma-Aldrich</td>
			<td>G9779&#160;</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">&#160;Glucose</td>
			<td>ThermoFisher Scientific</td>
			<td>15023-021</td>
			<td>25% (w/v) Solution, 0.2 mm filter sterilized</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Chicken Plasma</td>
			<td>Cocalico Biologicals</td>
			<td>30-0300-5L</td>
			<td>Rehydrate in sterile water, centrifuge at 2500 x g 30 min at 4 &#176;C, quick freeze aliquots in liquid nitrogen and store at&#160; -80 &#176;C.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Thrombin, Bovine</td>
			<td>Fisher</td>
			<td>60-516-01KU</td>
			<td>150 units /ml in GBSS/Glucose, quick freeze aliquots in liquid nitrogen and store at -80 &#176;C.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Cell Roller System</td>
			<td>Bellco Biotech</td>
			<td>SciERA</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Roller Incubator</td>
			<td>Forma</td>
			<td>Model 3956</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">N21-MAX</td>
			<td>ThermoFisher Scientific</td>
			<td>AR008</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Pen/Strep (100X)</td>
			<td>ThermoFisher Scientific</td>
			<td>15140122</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">200 mM Glutamine</td>
			<td>ThermoFisher Scientific</td>
			<td>25030081</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Glucose</td>
			<td>ThermoFisher Scientific</td>
			<td>15023-021</td>
			<td>25% (w/v) Solution, 0.2 mm filter sterilized</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Neurobasal A</td>
			<td>ThermoFisher Scientific</td>
			<td>10888-022&#160;</td>
			<td>Complete Medium: 48 mL Neurobasal A, 1 mL N21-MAX, 0.625 mL 200 mM Glutamine, 0.180 mL 25% Glucose, 0.250 mL 100x pen/strep.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Third generation lentivirus packaging</td>
			<td>Life Technologies</td>
			<td>K4975-00</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">159 K cutoff centrifugal filters (Centricon)</td>
			<td>EMD Millipore</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Lentiviral cloning system (InFusion)</td>
			<td>Clonetech</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Plasmids 30323, 50856, 51279</td>
			<td>Addgene</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Neuronal cell viability dye (NeuO)</td>
			<td>Stemcell technologies</td>
			<td>1801</td>
			<td>Thaw once and quick freeze in 4 &#181;L aliquots. Store at -20 &#176;C</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Inverted microscope</td>
			<td>Olympus</td>
			<td>IX83</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Microscope objectives</td>
			<td>Olympus</td>
			<td>air: 4X, 20; oil: 40X, 60X,</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Spinning disc confocal system</td>
			<td>Yokagawa</td>
			<td>CSU22</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Microscope EMCCD camera</td>
			<td>Photometrics</td>
			<td>Cascade II</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Linear encoded (x,y), piezo z flat top stage</td>
			<td>ASI</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Microscope lasers and integration</td>
			<td>Intelligent Imaging Innovations</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">HEK293T cells</td>
			<td>American Type Culture Collection</td>
			<td>CRL-3216</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Human Plasmin</td>
			<td>Sigma Aldrich</td>
			<td>P1867</td>
			<td>0.002 U/mL in 0.1% bovine serum albumin (0.2 mm filter sterilized), quick freeze in liquid nitrogen and store at -80 &#176;C.</td>
		</tr>
	</tbody>
</table>

modified roller tube method for precisely localized and repetitive intermittent imaging during long-term culture of brain slices in an enclosed system

Kultivert gnager hjernen stykker er nyttig for studerer cellulære og molekylære virkemåten til neurons og glia i et miljø som opprettholder mange av deres normale i vivo interaksjon. Skiver fra en rekke transgene musen linjer eller bruk av viral vektorer for uttrykk for fluorescently merket proteiner eller journalister i vill type hjernen skiver kan høy-resolution tenkelig ved fluorescens mikroskopi. Selv om flere metoder har blitt utviklet for imaging hjernen skiver, kombinerer skive kultur muligheten til å utføre utgjort repeterende høyoppløselige bilder i bestemte celler i levende skiver over lange tidsperioder problemer. Dette gjelder spesielt når viral vektorer brukes for uttrykk for eksogene proteiner siden dette er best gjort i et lukket system å beskytte brukerne og forhindre. Enkle endringer som er gjort til valse røret hjernen stykke kultur metoden, og at ved repeterende høyoppløselig avbildning av skiver over mange uker i et lukket system rapporteres. Dyrking skiver på photoetched coverslips tillater bruk av fiducial merker raskt og presist flytte scenen å bildet feltet identiske over tid før og etter ulike behandlinger. Eksempler vises for bruk av denne metoden kombinert med spesifikk neuronal farging og uttrykk for å observere endringer i hippocampus stykke arkitektur, viral-mediert neuronal uttrykk for fluorescerende proteiner og utvikling av cofilin patologi, som ble tidligere observert prefiks av Alzheimers sykdom (AD) skive behandling med oligomers av amyloid-β (Aβ) peptid som svar.

Fiducial merker kan brukes til å bestemme hvor nøyaktig for å reimage de samme cellene i de samme feltene over tid, undersøkte vi skiver dyrket på photoetched coverslips (figur 6en). Neurons var visualisert ved farging med en viktig fargestoff (100 nM for 2t; ikke flekker ikke-neuronal celler), som forsvinner fra neurons over tid uten å skade celler25. Vi identifisert en fiducial hake i en enkelt rute (<strong cl...

Watch this Scientific Journal Video about Modified Roller Tube Method for Precisely Localized and Repetitive Intermittent Imaging During Long-term Culture of Brain Slices in an Enclosed System at JoVE

Modified Roller Tube Method for Precisely Localized and Repetitive Intermittent Imaging During Long-term Culture of Brain Slices in an Enclosed System

Her er en modifisert valse røret metode for dyrking og intermitterende høyoppløselig avbilding av gnager hjernen skiver over mange uker med presis reposisjonering på photoetched coverslips. Neuronal levedyktighet og skive morfologi er godt vedlikeholdt. Anvendelser av innelukket systemet bruker virus for celle-type spesifikke uttrykk er gitt.

Modifisert Roller Tube metode for nettopp lokaliserte og repeterende intermitterende Imaging under langsiktige kultur på hjernen sektorer i et lukket System

Cultured rodent brain slices are useful for studying the cellular and molecular behavior of neurons and glia in an environment that maintains many of ...

Research

JoVE Journal

Neuroscience

A Polished and Reinforced Thinned-skull Window for Long-term Imaging of the Mouse Brain

En polert og forsterket tynnet-skalle Window for langsiktig Imaging av Mouse Brain

In vivo imaging of cortical function requires optical access to the brain without disruption of the intracranial environment. We present a method to form ...

A Molecular Readout of Long-term Olfactory Adaptation in C. elegans

A Molecular Readout of Long-term Olfactory Adaptation in C. elegans

En molekylær Avlesning av Long-term olfactory Tilpasning i<em&gt; C. elegans</em

During sustained stimulation most sensory neurons will adapt their response by decreasing their sensitivity to the signal. The adaptation response helps ...

Improved Preparation and Preservation of Hippocampal Mouse Slices for a Very Stable and Reproducible Recording of Long-term Potentiation

Forbedret Utarbeidelse og Bevaring av hippocampus Mouse Slices for en meget stabil og reproduserbar Recording of Long-term Potensiering

Long-term potentiation (LTP) is a type of synaptic plasticity characterized by an increase in synaptic strength and believed to be involved in memory ...

In vivo Postnatal Electroporation and Time-lapse Imaging of Neuroblast Migration in Mouse Acute Brain Slices

In vivo Postnatal Electroporation and Time-lapse Imaging of Neuroblast Migration in Mouse Acute Brain Slices

In vivo Postnatal Electroporation og Time-lapse Imaging av neuroblast Migration in Mouse Akutt Brain Slices

The subventricular zone (SVZ) is one of the main neurogenic niches in the postnatal brain. Here, neural progenitors proliferate and give rise to ...

Long-term Time Lapse Imaging of Mouse Cochlear Explants

Langsiktig Time Lapse Imaging av Mouse Cochlea eksplantater

Here we present a method for long-term time-lapse imaging of live embryonic mouse cochlear explants. The developmental program responsible for building ...

Regioselective Biolistic Targeting in Organotypic Brain Slices Using a Modified Gene Gun

Regioselektiv Biolistic Targeting i organotypic Brain Slices Ved hjelp av en modifisert Gene Gun

Transfection of DNA has been invaluable for biological sciences and with recent advances to organotypic brain slice preparations, the effect of various ...

Isolation, Culture and Long-Term Maintenance of Primary Mesencephalic Dopaminergic Neurons From Embryonic Rodent Brains

Isolasjon, Kultur og langsiktig vedlikehold av Primary mesencefalisk dopaminerge nevroner Fra Embryonic Gnagere Brains

Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson&#8217;s diseae. Study of the biological processes ...

Agarose Microchambers for Long-term Calcium Imaging of Caenorhabditis elegans

Agarose Microchambers for Long-term Calcium Imaging of Caenorhabditis elegans

Agaroseplater Microchambers for Long-term Kalsium Imaging av<em&gt; Caenorhabditis elegans</em

Behavior is controlled by the nervous system. Calcium imaging is a straightforward method in the transparent nematode Caenorhabditis elegans to measure ...

A Simplified Method for Ultra-Low Density, Long-Term Primary Hippocampal Neuron Culture

En forenklet metode for Ultra-Low Density, Long-Term Primær hippocampus Neuron Kultur

Culturing primary hippocampal neurons in vitro facilitates mechanistic interrogation of many aspects of neuronal development. Dissociated embryonic ...

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method

Utarbeidelse av akutt hjernen skiver med en optimalisert N-Methyl-D-glucamine beskyttende gjenopprettingsmetode

This protocol is a practical guide to the N-methyl-D-glucamine (NMDG) protective recovery method of brain slice preparation. Numerous recent studies have ...