Whole Mount Imaging: Ophthalmologic and Retina Studies

The retina is a vital sheet of nerve tissue at the back of the eye, transforming light signals into electrical impulses that are interpretable by the brain. A large amount of information, including synaptic connections, the spatial arrangement of neuronal arrays, electrophysiological properties of photoreceptors as well as blood vessel morphology and pathology can be easily analyzed by studying the retina as a whole mount or a flattened sheet of tissue.

As opposed to sectioning the whole eye tissue and getting cross-sections of small areas of retina, the whole mount technique involves dissecting out the retinal sheet and mounting it in a clover shape on a flat surface. This can be easily achieved by a few simple steps: (an example video can be found here)

  1. Euthanize the animal and enucleate the eyes. Fix the eyes in fixative at 4°C if necessary.
  2. Remove the cornea and pull out the lens using micro-forceps.
  3. Remove the vitreous and detach the retina from the eye cup.
  4. Perform standard immunohistochemistry on free-floating retina if necessary.
  5. Wash retinas in PBS and make four incisions in each quarter of the retina with a micro-scissor and spread the eyecup open like a flower on a glass slide.
  6. Add mounting media and coverslip.

That’s it! Now the entire retina can be imaged both in 2D or in 3D, which involves confocal imaging across a series of z steps. At Visikol, we excel in confocal image acquisition and image analysis, and we use deep learning to perform high-throughput and unbiased retinal blood vessel segmentation and quantification. In addition, we perform immunohistochemistry and confocal imaging on postmortem retina samples to examine vasculature as well as neuronal morphology in high resolution, allowing for higher resolution studies of retina pathology.

At Visikol, we offer tissue sample processing, mounting, and histological staining of tissues for ophthalmology research as well as neuroscience. We specialize in multiplexed immunohistochemistry and high content confocal imaging as well. Furthermore, with our expertise in deep learning-based image analysis, we conduct blood vessel quantification on in vivo images obtained via angiography. In addition, we are currently developing in vitro retinal coculture models as well as retinal organoids to facilitate ophthalmology research and drug discovery. If you are interested in the retina or ophthalmology research, please reach out to us! We are always interested to work together with our clients to develop customized assays to best suit their needs.

Services offered:

  • Wet tissue processing: fixation, processing, mounting
  • Histology of retina tissues
  • Immunohistochemistry / immunofluorescent labeling and imaging of choroidal flat mounts, retina mounts
  • Quantitative analysis of blood vessel area/volume, analysis of vessel regression
  • Quantitative analysis of FITC-angiography images
  • Quantitative analysis of blood vessel leakage

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