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Related Concept Videos

Flow Cytometry01:23

Flow Cytometry

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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Structures of Solids02:22

Structures of Solids

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Solids in which the atoms, ions, or molecules are arranged in a definite repeating pattern are known as crystalline solids. Metals and ionic compounds typically form ordered, crystalline solids. A crystalline solid has a precise melting temperature because each atom or molecule of the same type is held in place with the same forces or energy. Amorphous solids or non-crystalline solids (or, sometimes, glasses) which lack an ordered internal structure and are randomly arranged. Substances that...
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Metallic Solids02:37

Metallic Solids

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Metallic solids such as crystals of copper, aluminum, and iron are formed by metal atoms. The structure of metallic crystals is often described as a uniform distribution of atomic nuclei within a “sea” of delocalized electrons. The atoms within such a metallic solid are held together by a unique force known as metallic bonding that gives rise to many useful and varied bulk properties.
All metallic solids exhibit high thermal and electrical conductivity, metallic luster, and malleability....
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Network Covalent Solids02:18

Network Covalent Solids

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Network covalent solids contain a three-dimensional network of covalently bonded atoms as found in the crystal structures of nonmetals like diamond, graphite, silicon, and some covalent compounds, such as silicon dioxide (sand) and silicon carbide (carborundum, the abrasive on sandpaper). Many minerals have networks of covalent bonds.
To break or to melt a covalent network solid, covalent bonds must be broken. Because covalent bonds are relatively strong, covalent network solids are typically...
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Molecular and Ionic Solids02:54

Molecular and Ionic Solids

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Crystalline solids are divided into four types: molecular, ionic, metallic, and covalent network based on the type of constituent units and their interparticle interactions.
Molecular Solids
Molecular crystalline solids, such as ice, sucrose (table sugar), and iodine, are solids that are composed of neutral molecules as their constituent units. These molecules are held together by weak intermolecular forces such as London dispersion forces, dipole-dipole interactions, or hydrogen bonds, which...
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Molecular Comparison of Gases, Liquids, and Solids02:26

Molecular Comparison of Gases, Liquids, and Solids

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Particles in a solid are tightly packed together (fixed shape) and often arranged in a regular pattern; in a liquid, they are close together with no regular arrangement (no fixed shape); in a gas, they are far apart with no regular arrangement (no fixed shape). Particles in a solid vibrate about fixed positions (cannot flow) and do not generally move in relation to one another; in a liquid, they move past each other (can flow) but remain in essentially constant contact; in a gas, they move...
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Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry
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Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry

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Best Practices for Preparing a Single Cell Suspension from Solid Tissues for Flow Cytometry.

Andrew Reichard1, Kewal Asosingh1,2

  • 1Department of Inflammation and Immunity, Lerner Research Institute, The Cleveland Clinic, Cleveland, Ohio.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|December 8, 2018
PubMed
Summary
This summary is machine-generated.

Preparing single cell suspensions for flow cytometry requires careful tissue processing. This guide details optimizing digestion and dissociation to ensure cell viability and antigen integrity for accurate results.

Keywords:
disaggregationflow cytometrysingle cell suspensiontissue digestion

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Area of Science:

  • Immunology
  • Cell Biology
  • Biotechnology

Background:

  • Flow cytometry is a powerful technique for analyzing single cells, but requires high-quality single cell suspensions.
  • Solid tissues present challenges due to complex extracellular matrix and cell-cell junctions.
  • Proper sample preparation is crucial for reliable flow cytometry data.

Purpose of the Study:

  • To provide a general guide for preparing single cell suspensions from solid tissues for flow cytometry.
  • To highlight critical steps and considerations in tissue processing for optimal cell isolation.
  • To ensure cell viability and antigen preservation during dissociation.

Main Methods:

  • Tissue dissection and mechanical dissociation.
  • Enzymatic digestion to degrade the extracellular matrix.
  • Evaluation of single cell suspension quality (viability, debris, aggregates).

Main Results:

  • Successful isolation of single cells from solid tissues is achievable through optimized protocols.
  • Careful consideration of protein and matrix components is necessary for protocol design.
  • High cell viability and minimal debris are critical for downstream flow cytometry analysis.

Conclusions:

  • Optimized tissue processing is essential for generating high-quality single cell suspensions for flow cytometry.
  • The presented guide offers key steps for researchers preparing solid tissues for flow cytometry.
  • Adherence to proper protocols ensures reliable experimental outcomes in flow cytometry.