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Related Concept Videos

Imaging Biological Samples with Optical Microscopy01:18

Imaging Biological Samples with Optical Microscopy

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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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Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
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Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass, polymers, and biological samples. AFM offers over 1000 times more resolution than the optical imaging system. Images generated from AFM are three-dimensional surface profiles, offering an advantage over the flat, two-dimensional images from other imaging techniques.
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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Related Experiment Video

Updated: Aug 7, 2025

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip
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An integrated platform for high-throughput nanoscopy.

Andrew E S Barentine1,2, Yu Lin1,2, Edward M Courvan1,3

  • 1Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA.

Nature Biotechnology
|March 14, 2023
PubMed
Summary
This summary is machine-generated.

A new platform dramatically increases single-molecule localization microscopy throughput to 10,000 cells daily by integrating data compression and distributed analysis. This overcomes data bottlenecks for high-resolution 3D fluorescence imaging.

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Biomolecular Imaging of Cellular Uptake of Nanoparticles using Multimodal Nonlinear Optical Microscopy
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Area of Science:

  • Biophysics
  • Microscopy
  • Computational Biology

Background:

  • Single-molecule localization microscopy (SMLM) achieves nanoscale resolution in 3D fluorescence imaging.
  • High-resolution SMLM requires extensive camera frames, limiting throughput to tens of cells per day.
  • Increasing camera frame rates exacerbates data volume limitations in current SMLM workflows.

Purpose of the Study:

  • To develop an integrated acquisition and analysis platform for high-throughput SMLM.
  • To overcome data volume limitations and increase SMLM processing speed.
  • To enable automated, remote, and feedback-controlled SMLM acquisition and analysis.

Main Methods:

  • Implementation of microscopy-specific data compression techniques.
  • Utilizing distributed storage and distributed analysis for parallel processing.
  • Development of a graphically reconfigurable analysis pipeline within the PYthon-Microscopy Environment (PYME).

Main Results:

  • Achieved an acquisition and analysis throughput of 10,000 cells per day.
  • Demonstrated imaging of hundreds of cells per well in multi-well formats.
  • Platform facilitates automated analysis initiation during acquisition and remote execution.

Conclusions:

  • The integrated platform significantly enhances SMLM throughput, enabling large-scale cellular imaging.
  • PYME-based platform offers flexibility for custom microscopes and user-defined extensions.
  • This advancement facilitates high-throughput 3D super-resolution microscopy for biological research.