Name: in vivo imaging of the mouse spinal cord using two-photon microscopy
Uploaded: 2012-01-05T13:00:00
Description: Watch this Scientific Journal Video about In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy at JoVE.com

Dette arbejde blev støttet af National Scleroseforeningen give RG4595A1 / T til DD og NIH / NINDS stipendier NS051470, NS052189 og NS066361 til KA Tal og film tilpasset og / eller genoptrykt fra Davalos et al., J Neurosci Metoder. 2008 Marts 30, 169 (1) :1-7 Copyright 2008, med tilladelse fra Elsevier.

1. Opbygning af spinal stabilisering enhed <ol><li> Bestil Narishige STS-A Compact Rygmarv Klemmer og Narishige MA-6N hovedet holder adapter. </li><li> Brugerdefineret design og gøre en rustfri stål base plade til at holde de to Narishige dele i tilpasningen, således at dyrets hoved er understøttet, mens dens rygsøjlen og hale er fastspændt. Husk, at hele enheden skal passe under lup linsen som regel på en sænket mikroskop scene. </li></ol> 2. Animal kirurgi <ol><li> Pre-varme mikrosk...

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Metoden beskrevet her giver mulighed for stabile og gentagne in vivo billeddannelse af tætbefolkede fluorescerende cellulære strukturer i rygmarv fra bedøvet mus ved hjælp af to-foton mikroskopi. Den opnåede stabilitet er et resultat af et skræddersyet spinal stabilisering enhed og anæstesiregimet, som reducerer åndedræts-induceret bevægelse artefakt. Det spinal stabilisering enhed giver pusterum under musen kroppen og kan opbygges ved hjælp af kommercielt tilgængelige spinal klemmer og hoved monter...

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in vivo imaging of the mouse spinal cord using two-photon microscopy

In vivo imaging ved hjælp af to-foton mikroskopi 1 i mus, der er genetisk manipuleret til at udtrykke fluorescerende proteiner i bestemte celletyper 2-3 er blevet væsentligt udvidet vores viden om fysiologiske og patologiske processer i talrige væv in vivo 4-7. I studier af det centrale nervesystem (CNS), har der været en bred anvendelse af in vivo imaging i hjernen, som har produceret et væld af nye og ofte uventede resultater om opførslen af celler som neuroner, astrocytes, mikroglia, under fysiologiske eller patologiske tilstande 8-17. Imidlertid har det meste tekniske komplikationer begrænsede gennemførelse af in vivo billeddannelse i studier af den levende mus rygmarven. I særdeleshed genererer anatomiske nærhed af rygmarven til lungerne og hjertet betydelig bevægelse artefakt, der gør billedbehandling den levende rygmarven en udfordrende opgave. </p&gt; Vi har udviklet en ny metode, der overvinder de iboende begrænsninger i rygmarven imaging ved at stabilisere rygsøjlen, reducere luftvejs-inducerede bevægelser og dermed fremme brugen af to-foton mikroskopi til billede musen rygmarven in vivo. Dette opnås ved at kombinere en skræddersyet spinal stabilisering enhed med en metode til dyb anæstesi, hvilket resulterer i en betydelig reduktion af luftvejs-induceret bevægelser. Denne video-protokollen viser, hvordan du kan udsætte et lille område af den levende rygmarven, der kan vedligeholdes under stabile fysiologiske forhold over længere perioder ved at holde vævsskade og blødning til et minimum. Repræsentant rå billeder erhvervet in vivo detaljer i høj opløsning den tætte sammenhæng mellem mikroglia og kar. En timelapse sekvens viser den dynamiske adfærd microglial processer i levende mus rygmarven. Desuden en kontinuerlig scanning af samme z-frame demonstreres enestående stabilitet, at denne metode kan nå at generere stakke af billeder og / eller timelapse film, der ikke kræver billede styremærket-købet. Endelig vil vi vise, hvordan denne metode kan bruges til at vende tilbage til og Reimage det samme område af rygmarven på senere tidspunkter, der giver mulighed for longitudinelle studier af igangværende fysiologiske eller patologiske processer in vivo.

Watch this Scientific Journal Video about In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy at JoVE.com

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

Et minimalt invasiv protokol til at stabilisere musen rygsøjlen og udføre gentagne<em&gt; In vivo</em&gt; Rygmarven billeddannelse ved hjælp af to-foton mikroskopi er beskrevet. Denne metode kombinerer en spinal stabilisering enhed og anæstesiregimet at minimere åndedræts-inducerede bevægelser og producerer rå billeddata, som ikke kræver justering eller anden efterbehandling.

In vivo imaging af Mouse Spinal Cord ved hjælp af to-foton mikroskopi

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has ...

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Neuroscience

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

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Human Immunodeficiency Virus 1 (HIV-1) infection frequently results in HIV-1 Associated Neurocognitive Disorders (HAND), and is characterized by a chronic ...

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The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in ...

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Two-photon imaging is a powerful tool for the in vivo analysis of neuronal circuits in the mammalian brain. However, a limited number of in vivo imaging ...