Variable angle total internal reflection fluorescence microscopy in s-polarization: a new approach to quantify cell-substrate distances in contacts

Abdollah Hassanzadeh; Ata Bahmani; Silvia Mittler

doi:10.1117/12.872938

21 September 2010 Variable angle total internal reflection fluorescence microscopy in s-polarization: a new approach to quantify cell-substrate distances in contacts

Abdollah Hassanzadeh, Ata Bahmani, Silvia Mittler

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Proceedings Volume 7750, Photonics North 2010; 77500F (2010) https://doi.org/10.1117/12.872938
Event: Photonics North 2010, 2010, Niagara Falls, Canada

Abstract

Total internal reflection fluorescence microscopy is an evanescent based fluorescence microscope providing a selective visualization of cell-substrate contacts without interference from other, deeper cellular regions. Total internal reflection fluorescence microscope is used extensively to visualize cell-substrate contacts. However, quantifying these contacts - in particular the measurement of cell-substrate distances - has not been performed often. In order to quantify the cellsubstrate distances we have developed a new theoretical method which is based on a change in the penetration depth of the evanescent field by tuning the angle of incidence slightly above the angle of total internal reflection for s-polarized light. This is simpler and much more accurate in comparison to the few existing approaches.

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Abdollah Hassanzadeh, Ata Bahmani, and Silvia Mittler "Variable angle total internal reflection fluorescence microscopy in s-polarization: a new approach to quantify cell-substrate distances in contacts", Proc. SPIE 7750, Photonics North 2010, 77500F (21 September 2010); https://doi.org/10.1117/12.872938

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KEYWORDS

Luminescence

Distance measurement

Interfaces

Microscopy

Refractive index

Microscopes

Reflection

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