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To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam does not destroy them and dried thoroughly (desiccated/dehydrated) so the vacuum does not affect them. Fixation needs to be done as quickly as possible because the sample properties will start changing as soon as it is removed from its natural environment. For example, in a tissue sample, the oxygen levels begin decreasing, causing an altered...
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Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
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How to prepare cytological samples for molecular testing.

Claudio Bellevicine1, Umberto Malapelle1, Elena Vigliar1

  • 1Department of Public Health, Anatomic Pathology Unit, University of Napoli Federico II School of Medicine, Napoli, Italy.

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|July 26, 2017
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Summary

Standardizing molecular testing in cytopathology is crucial for accurate results. Optimizing preanalytical protocols and cytopathologist involvement ensures high-quality DNA and RNA for molecular diagnostics.

Keywords:
cancercell blockcytopathologydirect smearfine-needle aspirationmolecular cytopathologymutational analysisnext generation sequencingpersonalized medicinepolymerase chain reactionpreanalytic

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Area of Science:

  • Cytopathology
  • Molecular Diagnostics
  • Genomic Medicine

Background:

  • Cytological samples provide high-quality DNA suitable for molecular testing.
  • Molecular testing success hinges on standardized preanalytical protocols for DNA and RNA.
  • Cytopreparation significantly impacts RNA quality and reverse transcription-polymerase chain reaction (RT-PCR) outcomes.

Purpose of the Study:

  • To review challenges in standardizing and optimizing molecular testing workflows in cytopathology.
  • To highlight the importance of cytopathologist involvement in the molecular testing process.
  • To examine sample requirements for emerging molecular technologies.

Main Methods:

  • Literature review focusing on preanalytical variables in cytopathology molecular testing.
  • Discussion of the cytopathologist's role in specimen handling and quality control.
  • Examination of evolving technological demands on cytological samples.

Main Results:

  • Standardized preanalytical protocols are essential for maximizing DNA yield and quality.
  • RNA quality, critical for RT-PCR, is heavily influenced by cytopreparation techniques.
  • The cytopathologist's expertise is vital from test request to sample storage.

Conclusions:

  • Standardization of molecular testing workflows in cytopathology is a key challenge.
  • Enhanced collaboration and education for cytopathologists are necessary for successful molecular diagnostics.
  • Adapting to new technologies requires continuous professional development for cytopathologists.