Alternative Methods to Enhance the Axial Resolution of Total Internal Reflection Fluorescence–Structured Illumination Microscopy
Total internal reflection fluorescence–structured illumination microscopy (TIRF-SIM) can enhance the lateral resolution of fluorescence microscopy to twice the diffraction limit, enabling subtler observations of activity in subcellular life. However, the lack of an axial resolution makes it difficul...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-06-01
|
| Series: | Photonics |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2304-6732/12/7/652 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850071602361794560 |
|---|---|
| author | Xiu Zheng Xiaomian Cai Wenjie Liu Youhua Chen Cuifang Kuang |
| author_facet | Xiu Zheng Xiaomian Cai Wenjie Liu Youhua Chen Cuifang Kuang |
| author_sort | Xiu Zheng |
| collection | DOAJ |
| description | Total internal reflection fluorescence–structured illumination microscopy (TIRF-SIM) can enhance the lateral resolution of fluorescence microscopy to twice the diffraction limit, enabling subtler observations of activity in subcellular life. However, the lack of an axial resolution makes it difficult to resolve three-dimensional (3D) subcellular structures. In this paper, we present an alternative TIRF-SIM axial resolution enhancement method by exploiting quantitative information regarding the distance between fluorophores and the surface within the evanescent field. Combining the lateral super-resolution information of TIRF-SIM with reconstructed axial information, a 3D super-resolution image with a 25 nm axial resolution is achieved without attaching special optical components or high-power lasers. The reconstruction results of cell samples demonstrate that the axial resolution enhancement method for TIRF-SIM can effectively resolve the axial depth of densely structured regions. |
| format | Article |
| id | doaj-art-7bf37f7c6b6645a8b770d8de76d9ef2c |
| institution | DOAJ |
| issn | 2304-6732 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Photonics |
| spelling | doaj-art-7bf37f7c6b6645a8b770d8de76d9ef2c2025-08-20T02:47:17ZengMDPI AGPhotonics2304-67322025-06-0112765210.3390/photonics12070652Alternative Methods to Enhance the Axial Resolution of Total Internal Reflection Fluorescence–Structured Illumination MicroscopyXiu Zheng0Xiaomian Cai1Wenjie Liu2Youhua Chen3Cuifang Kuang4Ningbo Global Innovation Center, Zhejiang University, Ningbo 315100, ChinaNingbo Global Innovation Center, Zhejiang University, Ningbo 315100, ChinaCollege of Optical Science and Engineering, Zhejiang University, Hangzhou 310000, ChinaNingbo Global Innovation Center, Zhejiang University, Ningbo 315100, ChinaCollege of Optical Science and Engineering, Zhejiang University, Hangzhou 310000, ChinaTotal internal reflection fluorescence–structured illumination microscopy (TIRF-SIM) can enhance the lateral resolution of fluorescence microscopy to twice the diffraction limit, enabling subtler observations of activity in subcellular life. However, the lack of an axial resolution makes it difficult to resolve three-dimensional (3D) subcellular structures. In this paper, we present an alternative TIRF-SIM axial resolution enhancement method by exploiting quantitative information regarding the distance between fluorophores and the surface within the evanescent field. Combining the lateral super-resolution information of TIRF-SIM with reconstructed axial information, a 3D super-resolution image with a 25 nm axial resolution is achieved without attaching special optical components or high-power lasers. The reconstruction results of cell samples demonstrate that the axial resolution enhancement method for TIRF-SIM can effectively resolve the axial depth of densely structured regions.https://www.mdpi.com/2304-6732/12/7/652total internal reflection fluorescence microscopystructured illumination microscopyaxial resolution enhancementthree-dimensional super-resolution |
| spellingShingle | Xiu Zheng Xiaomian Cai Wenjie Liu Youhua Chen Cuifang Kuang Alternative Methods to Enhance the Axial Resolution of Total Internal Reflection Fluorescence–Structured Illumination Microscopy Photonics total internal reflection fluorescence microscopy structured illumination microscopy axial resolution enhancement three-dimensional super-resolution |
| title | Alternative Methods to Enhance the Axial Resolution of Total Internal Reflection Fluorescence–Structured Illumination Microscopy |
| title_full | Alternative Methods to Enhance the Axial Resolution of Total Internal Reflection Fluorescence–Structured Illumination Microscopy |
| title_fullStr | Alternative Methods to Enhance the Axial Resolution of Total Internal Reflection Fluorescence–Structured Illumination Microscopy |
| title_full_unstemmed | Alternative Methods to Enhance the Axial Resolution of Total Internal Reflection Fluorescence–Structured Illumination Microscopy |
| title_short | Alternative Methods to Enhance the Axial Resolution of Total Internal Reflection Fluorescence–Structured Illumination Microscopy |
| title_sort | alternative methods to enhance the axial resolution of total internal reflection fluorescence structured illumination microscopy |
| topic | total internal reflection fluorescence microscopy structured illumination microscopy axial resolution enhancement three-dimensional super-resolution |
| url | https://www.mdpi.com/2304-6732/12/7/652 |
| work_keys_str_mv | AT xiuzheng alternativemethodstoenhancetheaxialresolutionoftotalinternalreflectionfluorescencestructuredilluminationmicroscopy AT xiaomiancai alternativemethodstoenhancetheaxialresolutionoftotalinternalreflectionfluorescencestructuredilluminationmicroscopy AT wenjieliu alternativemethodstoenhancetheaxialresolutionoftotalinternalreflectionfluorescencestructuredilluminationmicroscopy AT youhuachen alternativemethodstoenhancetheaxialresolutionoftotalinternalreflectionfluorescencestructuredilluminationmicroscopy AT cuifangkuang alternativemethodstoenhancetheaxialresolutionoftotalinternalreflectionfluorescencestructuredilluminationmicroscopy |