Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Studying the Cytoskeleton01:17

Studying the Cytoskeleton

10.4K
The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...
10.4K
Overview of Electron Microscopy01:25

Overview of Electron Microscopy

16.2K
The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes can only magnify 1000X, and a few can magnify up to 1500X. Electrons, like electromagnetic radiation, can behave like waves, but with wavelengths of 0.005 nm, they produce significantly greater resolution up to 0.05 nm as compared to 500 nm for visible light. An electron microscope (EM) can create a sharp image that is magnified up to 2,000,000X.
16.2K
Overview of Microscopy Techniques01:22

Overview of Microscopy Techniques

17.6K
The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...
17.6K
Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

960
Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...
960
Microtubules in Signaling01:22

Microtubules in Signaling

2.2K
The primary cilium, made up of microtubules, acts as antennae on the cell surfaces for relaying external stimuli into the cells. These fine hair-like structures are present, generally one per cell. These are non-motile cilia in a 9+0 microtubules arrangement, where the central pair of microtubules are absent. The primary cilia arise from the basal body embedded in the cell membrane. Intraflagellar transport (IFT) carries requisite proteins from the cytoplasm to the cilium because the primary...
2.2K
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

21.7K
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,...
21.7K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Morphometric analysis of the thymic epithelial cell network using integrated and orthogonal digital pathology approaches.

Communications biology·2026
Same author

Mapping murine thymic epithelial cells: functional ultrastructure and implications for thymopoiesis.

Journal of leukocyte biology·2025
Same author

Uncovering synaptic and cellular nanoarchitecture of brain tissue via seamless <i>in situ</i> trimming and milling for cryo-electron tomography.

bioRxiv : the preprint server for biology·2025
Same author

Morphometric Analysis of the Thymic Epithelial Cell (TEC) Network Using Integrated and Orthogonal Digital Pathology Approaches.

bioRxiv : the preprint server for biology·2024
Same author

mTORC2-NDRG1-CDC42 axis couples fasting to mitochondrial fission.

Nature cell biology·2023
Same author

Alzheimer's-Associated Upregulation of Mitochondria-Associated ER Membranes After Traumatic Brain Injury.

Cellular and molecular neurobiology·2022
Same journal

Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

High-Performance Computing in Tandem Mass Spectrometry (MS/MS) Peptide Identification.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Engineering and Adapting Disulfide-Containing Proteins to Enable Intracellular Functionality.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

AI-Driven Protein Research: From Prediction to Design.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

Related Experiment Video

Updated: Mar 16, 2026

Simple Detection of Primary Cilia by Immunofluorescence
08:07

Simple Detection of Primary Cilia by Immunofluorescence

Published on: May 15, 2020

12.0K

CLEM Methods for Studying Primary Cilia.

Frank P Macaluso1, Geoffrey S Perumal1, Johan Kolstrup1

  • 1Analytical Imaging Facility, Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Forchheimer Building, Room 602, 1300 Morris Park Ave., Bronx, NY, 10461, USA.

Methods in Molecular Biology (Clifton, N.J.)
|August 13, 2016
PubMed
Summary
This summary is machine-generated.

Correlated light and electron microscopy (CLEM) precisely maps protein distribution in primary cilia. This technique combines fluorescence and scanning electron microscopy for detailed cellular structure and protein localization analysis.

Keywords:
AxonemeCLEMMEFPrimary ciliaSEMSuperresolution

More Related Videos

Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
11:16

Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles

Published on: August 7, 2016

10.3K
Correlative Light and Electron Microscopy CLEM as a Tool to Visualize Microinjected Molecules and their Eukaryotic Sub-cellular Targets
09:10

Correlative Light and Electron Microscopy CLEM as a Tool to Visualize Microinjected Molecules and their Eukaryotic Sub-cellular Targets

Published on: May 4, 2012

19.8K

Related Experiment Videos

Last Updated: Mar 16, 2026

Simple Detection of Primary Cilia by Immunofluorescence
08:07

Simple Detection of Primary Cilia by Immunofluorescence

Published on: May 15, 2020

12.0K
Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
11:16

Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles

Published on: August 7, 2016

10.3K
Correlative Light and Electron Microscopy CLEM as a Tool to Visualize Microinjected Molecules and their Eukaryotic Sub-cellular Targets
09:10

Correlative Light and Electron Microscopy CLEM as a Tool to Visualize Microinjected Molecules and their Eukaryotic Sub-cellular Targets

Published on: May 4, 2012

19.8K

Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Biophysics

Background:

  • Primary cilia are crucial cellular organelles involved in various signaling pathways.
  • Understanding protein localization within cilia is key to deciphering their function.
  • Correlated light and electron microscopy (CLEM) offers high-resolution imaging for biological samples.

Purpose of the Study:

  • To detail and compare methods for CLEM imaging of primary cilia.
  • To enable precise localization of proteins within ciliary structures.
  • To provide insights into ciliary protein distribution and function.

Main Methods:

  • Sequential fluorescence and scanning electron microscopy (SEM) processing for robust antibody localization.
  • Sample preparation for fluorescence imaging followed by SEM for optimal live-cell imaging.
  • Utilizing SEM for 3D measurements of ciliary length and orientation.

Main Results:

  • CLEM allows high-precision determination of tagged protein distribution along the ciliary membrane and axoneme.
  • SEM enables accurate measurement of ciliary dimensions and spatial relationships.
  • Combined superresolution microscopy can visualize ciliary component entry/exit points.

Conclusions:

  • CLEM is an effective technique for studying primary cilia structure and protein localization.
  • Different CLEM protocols offer advantages for specific research questions, such as antibody localization or live-cell imaging.
  • This approach enhances the understanding of primary cilia's role in cellular processes.