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Related Concept Videos

Imaging Biological Samples with Optical Microscopy01:18

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Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
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Robust adaptive optics for localization microscopy deep in complex tissue.

Marijn E Siemons1, Naomi A K Hanemaaijer1,2, Maarten H P Kole1,2

  • 1Cell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht University, Utrecht, the Netherlands.

Nature Communications
|June 8, 2021
PubMed
Summary
This summary is machine-generated.

We developed REALM, a new adaptive optics method, to overcome challenges in single-molecule localization microscopy (SMLM) for imaging deep within biological tissues. REALM significantly improves molecular organization visualization in complex samples.

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Area of Science:

  • Biophysics
  • Cell Biology
  • Microscopy

Background:

  • Single-molecule localization microscopy (SMLM) offers nanoscale resolution for studying molecular organization in cells.
  • Sample-induced aberrations in complex biological tissues hinder SMLM's application and accurate molecular localization.
  • Existing adaptive optics methods show inconsistent performance and limited aberration correction capabilities.

Purpose of the Study:

  • To systematically compare the performance of existing adaptive optics methods for SMLM.
  • To identify limitations in current adaptive optics approaches for biological imaging.
  • To develop and validate an improved adaptive optics method for enhanced SMLM in tissues.

Main Methods:

  • Systematic comparison of existing adaptive optics methods using simulations and standardized experimental samples.
  • Development of a novel adaptive optics method, REALM (Robust and Effective Adaptive Optics in Localization Microscopy).
  • REALM utilizes 297 frames of blinking molecules to correct aberrations up to 1 rad RMS.

Main Results:

  • Existing adaptive optics methods often provide limited aberration correction or introduce additional errors.
  • REALM demonstrates effective correction of significant aberrations.
  • REALM successfully improved single-molecule localization and enabled visualization of cytoskeletal spectrin organization.

Conclusions:

  • REALM represents a significant advancement in adaptive optics for SMLM.
  • The method enables deep-tissue imaging (50 μm) with improved resolution.
  • REALM overcomes key challenges in applying SMLM to complex biological samples.