Dual-detection confocal microscopy: high-speed surface profiling without depth scanning

Dong-Ryoung Lee; Dae-Gab Gweon; Hongki Yoo

doi:10.1117/12.2211893

9 March 2016 Dual-detection confocal microscopy: high-speed surface profiling without depth scanning

Dong-Ryoung Lee, Dae-Gab Gweon, Hongki Yoo

Proceedings Volume 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII; 97130F (2016) https://doi.org/10.1117/12.2211893
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

We propose a new method for three-dimensional (3-D) imaging without depth scanning that we refer to as the dual-detection confocal microscopy (DDCM). Compared to conventional confocal microscopy, DDCM utilizes two pinholes of different sizes. DDCM generates two axial response curves which have different stiffness according to the pinhole diameters. The two axial response curves can draw the characteristics curve of the system which shows the relationship between the axial position of the sample and the intensity ratio. Utilizing the characteristic curve, the DDCM reconstructs a 3-D surface profile with a single 2-D scanning. The height of each pixel is calculated by the intensity ratio of the pixel and the intensity ratio curve. Since the height information can be obtained directly from the characteristic curve without depth scanning, a major advantage of DDCM over the conventional confocal microscopy is a speed. The 3-D surface profiling time is dramatically reduced. Furthermore, DDCM can measure 3-D images without the influence of the sample condition since the intensity ratio is independent of the quantum yield and reflectance. We present two types of DDCM, such as a fluorescence microscopy and a reflectance microscopy. In addition, we extend the measurement range axially by varying the pupil function. Here, we demonstrate the working principle of DDCM and the feasibility of the proposed methods.

Citation Download Citation

Dong-Ryoung Lee, Dae-Gab Gweon, and Hongki Yoo "Dual-detection confocal microscopy: high-speed surface profiling without depth scanning", Proc. SPIE 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII, 97130F (9 March 2016); https://doi.org/10.1117/12.2211893

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KEYWORDS

Confocal microscopy

3D image processing

Microscopy

Profiling

Sensors

Tissues

3D metrology

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