In Vivo Imaging of Cerebrospinal Fluid Transport through the Intact Mouse Skull using Fluorescence Macroscopy

Amanda  M Sweeney; Virginia Pl&#225;; Ting Du; Guojun Liu; Qian Sun; Sisi Peng; Benjamin  A. Plog; Benjamin  T. Kress; Xiaowei Wang; Humberto Mestre; Maiken Nedergaard

doi:10.3791/59774

In vivo Imaging del trasporto del fluido cerebrospinale attraverso il teschio del topo intatto ut...

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JoVE Journal Neuroscience

In Vivo Imaging of Cerebrospinal Fluid Transport through the Intact Mouse Skull using Fluorescence Macroscopy

In vivo Imaging del trasporto del fluido cerebrospinale attraverso il teschio del topo intatto utilizzando la macroscopia di fluorescenza

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06:22 min

July 29, 2019

DOI: 10.3791/59774-v

Amanda M Sweeney*¹, Virginia Plá*¹, Ting Du¹, Guojun Liu¹, Qian Sun¹, Sisi Peng¹, Benjamin A. Plog¹, Benjamin T. Kress¹, Xiaowei Wang¹, Humberto Mestre¹, Maiken Nedergaard^1,2

¹Center for Translational Neuromedicine,University of Rochester Medical Center, ²Center for Translational Neuromedicine,University of Copenhagen

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Overview

This study presents a novel transcranial optical imaging technique that enables real-time mesoscopic imaging of cerebrospinal fluid (CSF) transport in the cortex of live mice through an intact skull. This method allows for dynamic measurements of glymphatic transport with reduced costs compared to other imaging modalities.

Key Study Components

Area of Science

Neuroscience
Imaging techniques
Cerebrospinal fluid dynamics

Background

Glymphatic transport plays a critical role in brain metabolism and waste clearance.
Monitoring CSF dynamics is essential for understanding its impact on neurological health.
Traditional imaging methods can be invasive and costly.
New techniques are needed to study CSF flow in vivo with minimal invasiveness.

Purpose of Study

To develop an effective imaging technique for assessing CSF transport in vivo.
To facilitate dynamic observation of glymphatic function in research settings.
To simplify surgical procedures for installing imaging apparatus.

Methods Used

Transcranial optical imaging platform.
Live mice were used as the biological model for studying CSF dynamics.
The technique involves the installation of a cisterna magna cannula and a head plate.
Extensive pre-surgical and surgical procedures were detailed to ensure accurate placement.
CSF tracers were infused and imaged in real-time to observe transport processes.

Main Results

The fluorescent tracer was observed in various CSF pools, demonstrating transport through the glymphatic system.
Imaging revealed that glymphatic influx was reduced following traumatic brain injury.
Quantitative analysis indicated a significant decrease in tracer uptake in the injured hemisphere.
The technique provides robust data for understanding CSF dynamics in health and disease.

Conclusions

This imaging technique significantly advances the study of glymphatic transport, enabling more accurate assessments of CSF dynamics.
The method's simplicity and cost-effectiveness expand its applicability in neuroscience research.
The findings enhance our understanding of CSF function and its implications for various neurological conditions.

Frequently Asked Questions

What are the advantages of the transcranial optical imaging technique?

The technique allows for real-time imaging of CSF transport with minimal invasiveness, making it more accessible and cost-effective compared to traditional imaging methods.

How is the cisterna magna cannula installed?

The cannula is implanted using simple surgical procedures that involve making a midline skin incision and exposing the skull. This process is minimally invasive after proper training.

What data is obtained using this imaging method?

The method provides dynamic imaging data on CSF tracer distribution, allowing researchers to analyze glymphatic flow patterns and responses to injuries.

How can this method be applied in future studies?

This technique can be adapted to study CSF dynamics in various neurological diseases, particularly those affecting glymphatic function.

What are the limitations of the technique?

While minimally invasive, the technique requires precise surgical skill and may need optimization for different animal models or conditions.

L'imaging ottico transcranico consente l'imaging ad ampio campo del trasporto di fluidi cerebrospinali nella corteccia di topi vivi attraverso un cranio intatto.

Questa nuova tecnica per l'imaging mesoscopico in tempo reale di tracce florescenti di CSF attraverso il cranio intatto di topi vivi può essere utilizzata per valutare il trasporto linfomanico. Il vantaggio principale è che questa tecnica consente misurazioni dinamiche di traccianti intracisternali in vivo a una frazione del costo di altre modalità di imaging. Le procedure necessarie per posizionare la piastra della testa sulla cisterna magna cannula sono semplici e minimamente invasive ma richiedono una pratica corretta.

La dimostrazione visiva dei passaggi critici di questo metodo è necessaria per massimizzare la qualità dell'immagine e consentire confronti riproducibili tra topi diversi e gruppi sperimentali. Dopo aver confermato una mancanza di risposta al dito del piedi bagnato il collo e la testa con acqua sterile e radere la pelliccia inumidita. Pulire la pelle esposta con un tampone alcolico per rimuovere eventuali capelli residui e posizionare il mouse in un telaio stereotattico sopra un tampone a temperatura controllata.

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