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Measuring lysosomal size and frequency by electron microscopy.

Michael W Hess1, Lukas A Huber2

  • 1Institute of Histology and Embryology, Medical University of Innsbruck, Innsbruck, Austria.

Methods in Cell Biology
|July 6, 2021
PubMed
Summary
This summary is machine-generated.

This study details electron microscopy methods for analyzing cell organelle size and quantity. These techniques precisely quantify lysosomes and autophagic organelles, aiding in understanding cell health and disease.

Keywords:
AutophagolysosomeCryofixationEndolysosomeHigh-pressure freezingLysosomeMorphometryUltrastructure

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Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Quantitative Analysis

Background:

  • Organelle dynamics, including size and abundance of late endocytic and autophagic compartments, reflect cellular physiological and pathological states.
  • Electron microscopy (EM) offers high-resolution imaging crucial for visualizing subcellular structures and immunocytochemical details.
  • Quantitative morphometry using EM, particularly with advanced cryopreparation, enables precise analysis of organelles.

Purpose of the Study:

  • To provide detailed protocols for preparing diverse biological specimens for electron microscopy.
  • To present methods for identifying and characterizing specific classes or subpopulations of lysosomes and related organelles.
  • To describe straightforward manual measurement techniques for globular or spheroid-shaped organelles.

Main Methods:

  • Step-by-step protocols for various sample preparation techniques suitable for electron microscopy.
  • Methodologies for distinguishing and categorizing lysosomes and related organelles.
  • Manual measurement procedures for quantifying the size of spherical organelles.

Main Results:

  • Established protocols for cryopreparation and sample processing for diverse specimens.
  • Defined criteria and tools for identifying and classifying lysosome and autophagosome subpopulations.
  • Validated manual measurement techniques for accurate morphometric analysis of organelles.

Conclusions:

  • The described protocols facilitate precise quantitative analysis of late endocytic and autophagic organelles using electron microscopy.
  • These methods enhance the understanding of cellular conditions by enabling accurate organelle morphometry.
  • The protocols are applicable to a wide range of specimens and offer straightforward manual measurement options.