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Neuroscience

Thinned-skull Cortical Window Technique for In Vivo Optical Coherence Tomography Imaging

Published: November 19th, 2012

DOI:

10.3791/50053

1Division of Biomedical Sciences, University of California, Riverside , 2Department of Bioengineering, University of California, Riverside

We present a method of creating a thinned-skull cortical window (TSCW) in a mouse model for in vivo OCT imaging of the cerebral cortex.

Optical coherence tomography (OCT) is a biomedical imaging technique with high spatial-temporal resolution. With its minimally invasive approach OCT has been used extensively in ophthalmology, dermatology, and gastroenterology1-3. Using a thinned-skull cortical window (TSCW), we employ spectral-domain OCT (SD-OCT) modality as a tool to image the cortex in vivo. Commonly, an opened-skull has been used for neuro-imaging as it provides more versatility, however, a TSCW approach is less invasive and is an effective mean for long term imaging in neuropathology studies. Here, we present a method of creating a TSCW in a mouse model for in vivo OCT imaging of the cerebral cortex.

Since its introduction in the early 1990's, OCT has been used extensively for biological imaging of tissue structure and function2. OCT generates cross-sectional images by measuring echo time delay of backscattered light4 by implementing low coherence light source with a fiber-optic Michelson interferometer2,4. SD-OCT, also known as Fourier domain OCT (FD-OCT), was first introduced in 19955 and offers a superior imaging modality compared with traditional time domain OCT (TD-OCT). In SD-OCT, the reference arm is kept stationary resulting in a high speed and ultra high resolution image acquisition6-9.

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1. Surgical Preparation

  1. Female CD 1 mice between the ages of 6-8 weeks were used in our experiments.
  2. Anesthetize the mouse with an intraperitoneal injection of a ketamine and xylazine combination (80 mg/kg ketamine/10 mg/kg xylazine). Place the mouse on a homeothermic pad to ensure optimal body temperature at ~37 °C. Continuously monitor level of anesthesia by testing animal's reflexes (e.g., pinching foot with blunt forceps) and inject more anesthesia when necessary.
  3. Lubricat.......

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After creating a thinned window over the cerebral cortex the vasculature should now be more visually prominent (Figure 1) and will allow for a deeper imaging depth (up to 1 mm). The right cortex is thinned to approximately 55 μm as compared to a normal skull measured at 140 μm (Figure 1) and provides greater optical clarity. Further thinning to 10-15 μm is possible11 however not necessary as the use of glass cover slips and skull plates are not implemented in our expe.......

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Imaging with OCT and a thinned-skull is a novel neuro-imaging technique that has only been recently investigated15, 16. In our experiments, we demonstrated the feasibility of SD-OCT imaging through a TSCW in a mouse model in vivo. From our results, the skull is thinned to approximately 55 μm and the penetration depth is obtained at approximately 1 mm with image resolution of 8 μm and 20 μm in the axial and lateral direction, respectively. In the signal intensity profile, OCT imaging through a.......

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This work was supported by the UC Discovery Proof of Concept grant and by the NIH (R00 EB007241). The authors would also like to thank Jacqueline Hubbard for her assistance in this experiment.

....

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Name Company Catalog Number Comments
Materials Company Catalogue number Comments
Ketamine Phoenix Pharmaceuticals 57319-542-02  
Xylazine Akorn, Inc. 139-236  
Artificial Tears Ointment Rugby 0536-6550-91  
Nair Church & Dwight Co., Inc. 4010130  
Sterile Alcohol Prep Pad Kendall Healthcare 6818  
Cotton Tipped Applicators Fisherbrand 23-400-115  
Betadine Solution Swabstick Purdue Products 67618-153-01  
Saline Solution, .9% Phoenix Pharmaceuticals 57319-555-08  
Stereotactic Frame Stoelting    
High Speed Surgical Hand Drill Foredom   38,000 rpm
Carbide Round Bur Stoelting   0.75 mm
Dura-Green Stones Shofu   Shank: HP
Shape: BA1
CompoMaster Coarse & CompoMaster Polisher Shofu   Shape: Mini-Pt.
SpaceDrapes Braintree Scientific, Inc.    

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