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Microtubules in Signaling

Microtubules in Signaling

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...

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Artificial Intelligence Approaches to Assessing Primary Cilia

Artificial Intelligence Approaches to Assessing Primary Cilia

Published on: May 1, 2021

Measuring the primary cilium length: improved method for unbiased high-throughput analysis.

Anneloes Dummer¹, Christian Poelma², Marco C DeRuiter¹

¹Department Anatomy and Embryology, Leiden University Medical Center, Leiden, The Netherlands.

|February 13, 2016

Summary

Accurately measuring primary cilium length is crucial for understanding cell signaling. The Pythagorean theorem (PyT) method offers a high-throughput, unbiased approach for measuring angled cilia, outperforming maximum intensity projection (MIP).

Keywords:

Cilium length Measurements Methods Primary cilia

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Area of Science:

Cell Biology
Biophysics
Microscopy

Background:

Primary cilia are vital cellular protrusions for mechanical and chemical sensing.
Cilia length impacts key signaling pathways like Hedgehog and TGFβ.
Existing methods for cilium length measurement are prone to bias and low throughput.

Purpose of the Study:

To develop and validate a standardized, high-throughput method for accurate primary cilium length measurement.
To compare the reliability of different measurement techniques, including Maximum Intensity Projection (MIP), Pythagorean Theorem (PyT), and 3D Alternative Angled Slicing (DAAS).

Main Methods:

Comparison of MIP, PyT, and DAAS for measuring straight and angled cilia in cultured cells.
Analysis of measurement accuracy based on cilium orientation relative to the imaging plane.
Intraclass correlation coefficients were used to assess agreement between methods.

Main Results:

MIP and DAAS accurately measured 'flat' cilia, with MIP offering higher throughput.
MIP significantly underestimated the length of angled cilia.
PyT and DAAS showed comparable results for angled cilia, with PyT being preferred due to higher throughput.

Conclusions:

The Pythagorean Theorem (PyT) is a universal, unbiased method for measuring straight primary cilia length.
PyT provides high-throughput measurements and is recommended for angled cilia.
Manual tracking remains suitable for irregularly shaped cilia.