Related Experiment Video

Updated: Aug 8, 2026

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins
06:43

Simultaneous Interference Reflection and Total Internal Reflection Fluorescence Microscopy for Imaging Dynamic Microtubules and Associated Proteins

Published on: May 3, 2022

Segmentation and 3D reconstruction of microtubules in total internal reflection fluorescence microscopy (TIRFM)

Stathis Hadjidemetriou1, Derek Toomre, James S Duncan

  • 1Departments of Diagnostic Radiology and Biomedical Engineering, New Haven, CT 06520, USA. stathis@noodle.med.yale.edu

Medical Image Computing and Computer-Assisted Intervention : MICCAI ... International Conference on Medical Image Computing and Computer-Assisted Intervention
|May 12, 2006
PubMed

Insights

We developed an automated method to analyze total internal reflection fluorescence microscopy (TIRFM) images of microtubules, enabling high-throughput quantitative studies of their function in cell processes.

Area of Science:

  • Cell biology
  • Microscopy
  • Biophysics

Background:

  • Microtubule-cortex interactions are vital for cellular functions like vesicle transport, cell motility, and mitosis.
  • Dysfunctional microtubules are implicated in cancer progression.
  • Total internal reflection fluorescence microscopy (TIRFM) is crucial for observing microtubule dynamics near the cell cortex.

Purpose of the Study:

  • To develop an automated method for processing TIRFM images of microtubules.
  • To enable high-throughput quantitative analysis of microtubule behavior.
  • To overcome limitations of manual analysis in TIRFM studies.

Main Methods:

  • Image processing algorithm to automatically extract microtubule segments from TIRFM data.
  • Utilizing Hamilton-Jacobi equations for segment description.
  • Performing limited 3D reconstruction of microtubules.

Main Results:

  • Successful automated processing of TIRFM images.
  • Quantitative analysis of microtubule segments.
  • Validation of the method using both phantom and real cell imaging data.

Conclusions:

  • The developed automated method significantly enhances the efficiency and throughput of TIRFM image analysis.
  • This approach facilitates high-throughput quantitative studies of microtubule-cortex interactions.
  • The method holds potential for advancing research in cell biology and cancer.

More Related Videos

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules09:45

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules

Published on: August 8, 2019

Simultaneous Visualization of the Dynamics of Crosslinked and Single Microtubules In Vitro by TIRF Microscopy07:20

Simultaneous Visualization of the Dynamics of Crosslinked and Single Microtubules In Vitro by TIRF Microscopy

Published on: February 18, 2022

Related Concept Videos

Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
Electron Microscope Tomography and Single-particle Reconstruction01:07

Electron Microscope Tomography and Single-particle Reconstruction

Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron microscope tomography and single-particle reconstruction. While single-particle reconstruction can examine macromolecules and macromolecular complexes in vitro conditions only, tomography permits the study of cell components or small cells in vivo.
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...
Three-Dimensional Microscopy in Microbiology01:28

Three-Dimensional Microscopy in Microbiology

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...