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Related Concept Videos

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Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...
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Rapid histological imaging of bone without microtome sectioning using nonlinear microscopy.

Tadayuki Yoshitake1, Seymour Rosen2, Lucas C Cahill3

  • 1Department of Electrical Engineering and Computer Science and Research Laboratory of Electronics, Massachusetts Institute of Technology, Cambridge, MA, USA.

Bone
|November 7, 2021
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Summary

Nonlinear microscopy (NLM) offers rapid, intraoperative evaluation of bone pathology, visualizing normal and diseased bone architecture. This technique shows promise for real-time diagnostics, though training is needed for accurate interpretation.

Keywords:
Histology of boneNonlinear microscopyRapid tissue examinationTwo photon excitation microscopy

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Area of Science:

  • Biomedical Optics
  • Histopathology
  • Skeletal Biology

Background:

  • Conventional bone histology is time-consuming, hindering intraoperative assessment.
  • Nonlinear microscopy (NLM) provides optical sectioning for fresh tissue imaging without microtome sectioning.

Purpose of the Study:

  • To develop and evaluate a rapid protocol for NLM imaging of bone and associated cartilage.
  • To compare NLM imaging with conventional paraffin histology for bone pathology evaluation.

Main Methods:

  • NLM imaging of 71 bone specimens (normal, arthritic, malignant, inflammatory) from 40 patients.
  • Rapid staining (5 min) with acridine orange and sulforhodamine 101, followed by NLM imaging.
  • Comparison of NLM images with subsequent paraffin-embedded hematoxylin and eosin (H&E) stained slides.

Main Results:

  • NLM visualized normal bone architecture (lamellar matrix, osteocytes) and pathological features (osteoarthritis, osteomyelitis, malignancy).
  • NLM images resembled H&E, with noted differences in cellular details requiring interpreter training.
  • Rapid 3D translation and sub-surface imaging capabilities of NLM mitigated challenges from tissue surface irregularities.

Conclusions:

  • NLM is a promising technique for rapid, real-time evaluation of bone pathology.
  • Further studies are warranted to assess the diagnostic performance of NLM for bone diseases.