Effective improvement of subtraction method for light sheet fluorescence microscopy via tangent-function subtraction coefficient

Van Nhu Le; Xiaona Wang; Cuifang Kuang; Xu Liu

doi:10.1117/12.2506484

15 November 2018 Effective improvement of subtraction method for light sheet fluorescence microscopy via tangent-function subtraction coefficient

Van Nhu Le, Xiaona Wang, Cuifang Kuang, Xu Liu

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Proceedings Volume 10964, Tenth International Conference on Information Optics and Photonics; 1096462 (2018) https://doi.org/10.1117/12.2506484
Event: Tenth International Conference on Information Optics and Photonics (CIOP 2018), 2018, Beijing, China

Abstract

Light sheet fluorescence microscopy (LSFM) with fluorescence emission difference (FED) obtains axial resolution-enhanced images by the subtraction of signals excited by different light sheets, Gaussian light sheet and negative light sheet. Negative values are inevitable in FED LSFM with conventional subtraction coefficient, due to mismatches in the outer contours of the raw signals excited by different light sheets, causing image distortion. In this paper, we propose a method based on tangent-function subtraction coefficient to reduce image distortion. This tangent-function subtraction coefficient is introduced to minimize image distortion without lowering the axial resolution. The value of the variable subtraction coefficient is obtained from the intensity value of raw signals. We performed experiments using fluorescent beads with a diameter of 1 micron and compared the subtraction results with constant and tangent-function subtraction coefficient. The experimental results demonstrated that the proposed tangent-function subtraction coefficient has an advantage over the conventional constant coefficient.

Citation Download Citation

Van Nhu Le, Xiaona Wang, Cuifang Kuang, and Xu Liu "Effective improvement of subtraction method for light sheet fluorescence microscopy via tangent-function subtraction coefficient", Proc. SPIE 10964, Tenth International Conference on Information Optics and Photonics, 1096462 (15 November 2018); https://doi.org/10.1117/12.2506484

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KEYWORDS

Point spread functions

Luminescence

Microscopy

Beam splitters

Field emission displays

Spatial light modulators

Distortion

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