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Related Concept Videos

Confocal Fluorescence Microscopy01:16

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Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
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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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Multiplexed laser particles for spatially resolved single-cell analysis.

Sheldon J J Kwok1,2,3, Nicola Martino1, Paul H Dannenberg1,2

  • 11Harvard Medical School and Wellman Center for Photomedicine, Massachusetts General Hospital, 50 Blossom Street, Boston, MA 02114 USA.

Light, Science & Applications
|October 25, 2019
PubMed
Summary
This summary is machine-generated.

Single-cell analysis in tissues is crucial for understanding development and disease. Novel laser particles offer a high-throughput method to tag cells in situ for comprehensive downstream molecular analysis, preserving spatial information.

Keywords:
BiophotonicsImaging and sensingMicroresonators

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Genomics

Background:

  • Single-cell analysis is vital for studying cellular heterogeneity in development and diseases like cancer.
  • Current methods often require tissue dissociation, losing spatial context and limiting throughput.
  • Existing in situ techniques have limitations in the number and type of molecules analyzed or the number of cells profiled.

Purpose of the Study:

  • To review current methods for spatially resolved single-cell analysis.
  • To discuss the potential of novel multiplexed imaging probes (laser particles) for high-throughput analysis.
  • To address the need for methods that seamlessly integrate spatial mapping with single-cell techniques.

Main Methods:

  • Review of existing in situ multiplexed imaging and low-throughput ex vivo single-cell analysis techniques.
  • Discussion of emerging laser particle technology for in situ cell tagging.
  • Exploration of high-throughput single-cell molecular analysis.

Main Results:

  • Current methods for spatially resolved single-cell analysis are limited in scope and throughput.
  • Tissue dissociation leads to loss of crucial spatial information.
  • Laser particles present a promising approach for high-throughput, spatially resolved single-cell analysis.

Conclusions:

  • There is a critical need for high-throughput methods that maintain spatial information in single-cell analysis.
  • Laser particles offer a potential solution for comprehensive, spatially resolved single-cell profiling.
  • This technology could advance the study of cellular heterogeneity in complex biological systems.