Home › Microscopy & Imaging › High-resolution Episcopic Microscopy (HREM) - Simple and Robust Protocols for Processing and Visualizing Organic Materials
Microscopy & Imaging JoVE (Open Access) Citable · DOI

High-resolution Episcopic Microscopy (HREM) - Simple and Robust Protocols for Processing and Visualizing Organic Materials

DOI: 10.3791/56071-v
What you'll learn
  • ✓ Prepare embryonic and adult tissue samples for high-resolution episcopic microscopy
  • ✓ Embed biological material in resin using robust processing protocols
  • ✓ Acquire and visualize high-resolution digital volume data from organic tissues
  • ✓ Generate 3D reconstructions of embryos and tissue samples
Protocol

We provide simple and robust protocols for processing biopsy material of various species, embryos of biomedical model organisms and samples of other organic tissues in order to permit digital volume data generation with the high-resolution episcopic microscopy method.

Difficulty
intermediate
Total time
~3-5 days per sample (including tissue preparation, embedding, and imaging)
Model organism
Mouse embryos, zebrafish, other biomedical model organisms
Biosafety
BSL-1

Steps

1
Prepare embryonic and biopsy tissues

Extract and prepare embryonic tissues from model organisms or biopsy material from various species. Follow appropriate fixation and initial processing steps to preserve tissue morphology.

▶ 01:33
2
Embed tissues in resin medium

Infiltrate prepared samples with embedding resin and cure using appropriate protocols. Ensure uniform embedding to permit sectioning and imaging.

▶ 02:58
3
Acquire high-resolution episcopic microscopy images

Set up and operate high-resolution episcopic microscopy to capture serial optical sections of embedded tissue blocks. Generate digital volume data from sequential image acquisition.

▶ 04:36
4
Visualize and reconstruct embryo and tissue data

Process acquired image data to generate 3D reconstructions and visualizations of embryonic and adult tissue samples. Demonstrate visualization outcomes from complete protocol.

▶ 05:52
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