Resonant dielectric multilayer with controlled absorption for enhanced total internal reflection fluorescence microscopy

Total internal reflection fluorescence microscopy (TIRF-M) is widely used in biological imaging. Evanescent waves, generated at the glass-sample interface, theoretically strongly improve the axial resolution down to a hundred of nanometers. However, objective based TIRF-M suffers from different limi...

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Bibliographic Details
Main Authors: Favard, C. (Author), Koc, C. (Author), Lemarchand, F. (Author), Lereu, A.L (Author), Lumeau, J. (Author), Moreau, A. (Author), Mouttou, A. (Author)
Format: Article
Language:English
Published: NLM (Medline) 2022
Subjects:
Online Access:View Fulltext in Publisher
LEADER 02317nam a2200277Ia 4500
001 10.1364-OE.457353
008 220510s2022 CNT 000 0 und d
020 |a 10944087 (ISSN) 
245 1 0 |a Resonant dielectric multilayer with controlled absorption for enhanced total internal reflection fluorescence microscopy 
260 0 |b NLM (Medline)  |c 2022 
856 |z View Fulltext in Publisher  |u https://doi.org/10.1364/OE.457353 
520 3 |a Total internal reflection fluorescence microscopy (TIRF-M) is widely used in biological imaging. Evanescent waves, generated at the glass-sample interface, theoretically strongly improve the axial resolution down to a hundred of nanometers. However, objective based TIRF-M suffers from different limitations such as interference fringes and uneven illumination, mixing both propagating and evanescent waves, which degrade the image quality. In principle, uneven illumination could be avoided by increasing the excitation angle, but this results in a drastic loss of excitation power. We designed dedicated 1D photonic crystals in order to circumvent this power loss by directly acting on the intensity of the evanescent field at controlled incident angles. In this framework, we used dedicated resonant multi-dielectric stacks, supporting Bloch surface waves and resulting in large field enhancement when illuminated under the conditions of total internal reflection. Here, we present a numerical optimization of such resonant stacks by adapting the resulting resonance to the angular illumination conditions in TIRF-M and to the fluorescence collection constraints. We thus propose a dedicated resonant structure with a control of the absorption during thin film deposition. A first experimental demonstration illustrates the concept with a 3-fold fluorescence enhancement in agreement with the numerical predictions. 
650 0 4 |a article 
650 0 4 |a excitation 
650 0 4 |a illumination 
650 0 4 |a prediction 
650 0 4 |a thin film (procedure) 
650 0 4 |a total internal reflection fluorescence microscopy 
700 1 |a Favard, C.  |e author 
700 1 |a Koc, C.  |e author 
700 1 |a Lemarchand, F.  |e author 
700 1 |a Lereu, A.L.  |e author 
700 1 |a Lumeau, J.  |e author 
700 1 |a Moreau, A.  |e author 
700 1 |a Mouttou, A.  |e author 
773 |t Optics express