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Related Concept Videos

Preparation of Samples for Electron Microscopy01:20

Preparation of Samples for Electron Microscopy

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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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Sample Preparation for Analysis: Overview01:21

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Sample preparation is an essential step in the analytical process. It involves preparing a sample so that it can be analyzed accurately. The goal is to extract the analyte, the substance you want to measure, from the sample while removing any components that may interfere with the analysis. Sample preparation techniques vary depending on the physical state of the sample.
Bulk or large solid samples are typically reduced in size using grinding, crushing, or milling techniques to increase the...
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Sample Preparation for Analysis: Advanced Techniques01:08

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Accurate analysis of complex samples often requires advanced preparation techniques to achieve reliable and reproducible results. Samples containing inorganic or organic materials can be challenging to dissolve or decompose effectively. Standard sample preparation methods include acid digestion, fusion, dry ashing, and wet digestion.
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Imaging Biological Samples with Optical Microscopy01:18

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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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Sampling Methods: Sample Types01:18

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Sampling materials are classified into three main types: solid, liquid, and gas.
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Sampling Plans01:23

Sampling Plans

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Sampling is a crucial step in analytical chemistry, allowing researchers to collect representative data from a large population. Common sampling methods include random, judgmental, systematic, stratified, and cluster sampling.
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Optimizing Sample Preparation for Cryogenic Electron Microscopy
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Preparation of Diplonemid Samples for Microscopy.

Galina Prokopchuk1,2, Daria Tashyreva3, Orsola Iorillo1,2

  • 1Institute of Parasitology, Biology Centre, Czech Academy of Sciences, České Budějovice (Budweis), Czech Republic.

Methods in Molecular Biology (Clifton, N.J.)
|February 2, 2026
PubMed
Summary

This chapter details microscopy protocols for diplonemid cells, covering live and fixed imaging, endocytosis, immunofluorescence assay (IFA), and fluorescence in situ hybridization (FISH) for cellular biology research.

Keywords:
Cell fixationCell permeabilizationEndocytosis assayFluorescence in situ hybridizationFluorescence microscopyFluorescence stainingImmunofluorescence assayLive-cell imaging

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Area of Science:

  • Microscopy and cell biology

Background:

  • Diplonemids are marine stramenopiles with complex cellular structures that are challenging to study using traditional microscopy techniques.
  • Understanding diplonemid cell biology requires robust and adaptable protocols for high-resolution imaging.

Purpose of the Study:

  • To provide a comprehensive set of standardized protocols for preparing diplonemid cells for various microscopy applications.
  • To enable researchers to obtain high-quality imaging data for the cellular biology of diplonemids.

Main Methods:

  • Detailed protocols for slide preparation, cell harvesting, chemical fixation, and permeabilization.
  • Instructions for fluorescent labeling, including immunofluorescence assay (IFA) and fluorescence in situ hybridization (FISH).
  • Adaptable methods for live-cell imaging and fixed-cell fluorescence microscopy.

Main Results:

  • Established a suite of reliable protocols for diverse microscopy techniques.
  • Highlighted critical steps to ensure successful cell preparation and high-quality imaging.
  • Demonstrated the adaptability of protocols for different experimental objectives.

Conclusions:

  • These protocols facilitate advanced imaging of diplonemid cells, advancing the study of their cellular biology.
  • The standardized methods will aid researchers in exploring complex cellular processes in diplonemids.
  • High-quality imaging data is achievable through meticulous adherence to these preparation techniques.