Home › Analytical Chem › Where You Cut Matters: A Dissection and Analysis Guide for the Spatial Orientation of the Mouse Retina from Ocular Landmarks
Analytical Chem JoVE (Open Access) Citable · DOI

Where You Cut Matters: A Dissection and Analysis Guide for the Spatial Orientation of the Mouse Retina from Ocular Landmarks

DOI: 10.3791/57861-v
What you'll learn
  • ✓ Identify mouse retinal anatomical poles using ocular landmarks (superior rectus, choroid fissure)
  • ✓ Apply s-opsin immunohistochemistry to map cone photoreceptor gradients
  • ✓ Use Retistruct software and custom code to spatially orient isolated retinae
  • ✓ Analyze retinal dissection samples with anatomically accurate orientation
Protocol

Biopharma Insights This protocol provides a comprehensive dissection and analysis guide for the use of deep ocular landmarks, s-opsin immunohistochemistry, Retistruct, and custom code to accurately and reliably orient the isolated mouse retina in anatomical space.

Difficulty
advanced
Total time
~3–4 hours per retina (dissection, immunostaining, imaging, and analysis)
Model organism
Mouse (C57BL/6J or similar laboratory strain)
Biosafety
BSL-1

Steps

1
Identify superior rectus muscle landmark

Locate and use the superior rectus muscle as a deep ocular landmark to establish the dorsal pole and anatomical orientation of the mouse retina during initial dissection.

▶ 00:37
2
Identify choroid fissure landmark

Use the choroid fissure (ventral anatomical feature) as a secondary landmark to confirm retinal pole identification and establish ventral orientation in the isolated retina.

▶ 02:36
3
Perform s-opsin immunohistochemistry labeling

Apply s-opsin immunofluorescent staining to visualize short-wavelength cone photoreceptors and their dorsal-to-ventral gradient across the retinal surface.

▶ 04:06
4
Analyze retinal poles using s-opsin gradient

Combine s-opsin staining pattern with Retistruct software and custom code to computationally identify and confirm all four retinal poles (dorsal, ventral, nasal, temporal) in anatomical space.

▶ 07:02
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