Resolution Evaluation Method and Applications of 3D Microscopic Images

Xiliang Luo; Zhou Zhou; Jiangfeng Huang; Xiangjiang Dong; Gang Zheng; Ling Fu

doi:10.3788/CJL202249.0507205

Chinese Journal of Lasers, Volume. 49, Issue 5, 0507205(2022)

Resolution Evaluation Method and Applications of 3D Microscopic Images

Xiliang Luo^1,2, Zhou Zhou^1,2, Jiangfeng Huang^1,2, Xiangjiang Dong^1,2, Gang Zheng^3、*, and Ling Fu^1,2

¹Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China

²MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China

³School of Electronic Information and Communications, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China

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Figures & Tables(13)

Fig. 1. Flow chart of FRC resolution calculation

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Fig. 2. Working principle diagram of sFSC method’s shell selector. (a) Selector model parameters in spatial diagram；(b) double wedge symmetric selection model; (c) sFSC selector workflow simulation diagram at single frequency shell domain

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Fig. 3. Resolution evaluation results of fluorescent balls with 0.1 μm diameter. (a) Intensity profile of the fluorescent balls; (b) Gaussian fitting results of intensity profile by FWHM method; (c) angle-dependent resolution curve calculated by sFSC method

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Fig. 4. Flow chart of image restoration based on sFSC resolution evaluation method

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Fig. 5. Comparison of resolution and image quality before and after restoration. (a) Resolution curves using sFSC method；(b) image comparison before and after Wiener deconvolution; (c) intensity profile curves of ROI in Fig. 5(b)

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Fig. 6. Comparison of images reconstructed by Wiener deconvolution under different PSF. (a) Original data; (b) T-PSF; (c) R-PSF1; (d) R-PSF2; (e) O-PSF; (f) sFSC-PSF

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Fig. 7. Maximum projection images reconstructed by different deconvolution methods on different PSF size

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Fig. 8. sFSC resolution curves of LW deconvolution with R-PSF and sFSC-PSF input. (a) R-PSF (0.18 μm×0.55 μm)；(b) sFSC-PSF (0.198 μm×0.679 μm)

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Table 1. Comparison of resolution results among theoretical calculation, FWHM, and sFSC methods　unit：μm
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Table 1. Comparison of resolution results among theoretical calculation, FWHM, and sFSC methods　unit：μm
Method Lateral resolution Axial resolution
Theoretical 0.292 0.979
FWHM 0.294 1.297
sFSC 0.312 1.648

Table 2. BIBLE score of the maximum projection images reconstructed by Winner deconvolution on different PSF size
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Table 2. BIBLE score of the maximum projection images reconstructed by Winner deconvolution on different PSF size
PSF Original data T-PSF R-PSF1 R-PSF2 O-PSF sFSC-PSF
Size /(μm×μm) 0.204×0.458 0.18×0.55 0.22×0.60 0.25×0.60 0.198×0.679
Score 4.04 4.94 5.33 5.09 4.82 5.40

Table 3. sFSC resolution evaluation results of images reconstructed with Wiener deconvolution on different PSF size

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Table 3. sFSC resolution evaluation results of images reconstructed with Wiener deconvolution on different PSF size

PSF	Original data	T-PSF	R-PSF1	R-PSF2	O-PSF	sFSC-PSF
Size /(μm×μm)		0.204×0.458	0.18×0.55	0.22×0.60	0.25×0.60	0.198×0.679
Lateral resolution /μm	0.1984	0.1737	0.1707	0.1763	0.1814	0.1693
Axial resolution /μm	0.6792	0.4591	0.4610	0.4657	0.5167	0.4538

Table 4. BIBLE score of the maximum projection images reconstructed by different deconvolution methods on different PSF

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Table 4. BIBLE score of the maximum projection images reconstructed by different deconvolution methods on different PSF

PSF		T-PSF	R-PSF1	R-PSF2	O-PSF	sFSC-PSF
Size /(μm×μm)		0.204×0.458	0.18×0.55	0.22×0.60	0.25×0.60	0.198×0.679
BIBLE score	RL	3.94	4.18	3.78	3.56	4.38
	NNLS	2.85	2.95	2.68	2.50	3.22
	LW	2.86	3.19	2.83	2.66	3.38
	TRIF	2.94	3.36	2.93	2.70	3.51

Table 5. sFSC resolution evaluation results of images reconstructed by different deconvolution methods on different PSF

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Table 5. sFSC resolution evaluation results of images reconstructed by different deconvolution methods on different PSF

PSF		T-PSF	R-PSF1	R-PSF2	O-PSF	sFSC-PSF
Size /(μm×μm)		0.204×0.458	0.18×0.55	0.22×0.60	0.25×0.60	0.198×0.679
Lateral resolution /μm	RL	0.1941	0.1820	0.1986	0.2047	0.1725
	NNLS	0.1912	0.1851	0.1969	0.2126	0.1833
	LW	0.1813	0.1705	0.1904	0.2097	0.1824
	TRIF	0.1823	0.1665	0.1916	0.2125	0.1665
Axial resolution /μm	RL	0.6229	0.6178	0.6355	0.6409	0.6160
	NNLS	0.5362	0.5329	0.5283	0.5337	0.5307
	LW	0.5309	0.4999	0.5269	0.5381	0.5013
	TRIF	0.5265	0.5035	0.5208	0.5347	0.5011

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Xiliang Luo, Zhou Zhou, Jiangfeng Huang, Xiangjiang Dong, Gang Zheng, Ling Fu. Resolution Evaluation Method and Applications of 3D Microscopic Images[J]. Chinese Journal of Lasers, 2022, 49(5): 0507205

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Paper Information

Received: Dec. 6, 2021

Accepted: Jan. 11, 2022

Published Online: Mar. 9, 2022

The Author Email: Gang Zheng (ZhengGang@hust.edu.cn)

DOI:10.3788/CJL202249.0507205

Topics

laser devices and laser physics

Lasers and Laser Optics

Laser physics

laser manufacturing

Instrumentation, Measurement and Metrology

Table 1. Comparison of resolution results among theoretical calculation, FWHM, and sFSC methods unit：μm

Table 1. Comparison of resolution results among theoretical calculation, FWHM, and sFSC methods unit：μm

Table 2. BIBLE score of the maximum projection images reconstructed by Winner deconvolution on different PSF size

Table 2. BIBLE score of the maximum projection images reconstructed by Winner deconvolution on different PSF size

Table 3. sFSC resolution evaluation results of images reconstructed with Wiener deconvolution on different PSF size

Table 3. sFSC resolution evaluation results of images reconstructed with Wiener deconvolution on different PSF size

Table 4. BIBLE score of the maximum projection images reconstructed by different deconvolution methods on different PSF

Table 4. BIBLE score of the maximum projection images reconstructed by different deconvolution methods on different PSF

Table 5. sFSC resolution evaluation results of images reconstructed by different deconvolution methods on different PSF

Table 5. sFSC resolution evaluation results of images reconstructed by different deconvolution methods on different PSF

Table 1. Comparison of resolution results among theoretical calculation, FWHM, and sFSC methods　unit：μm

Table 1. Comparison of resolution results among theoretical calculation, FWHM, and sFSC methods　unit：μm