← Back to Volume 11

Lens-Free Microscopic Imaging for 3D Analysis of Insect Microstructures and Ecological Applications

Yangxiao (Olivia) Ou
pp.141-149 DOI: 10.62051/4esn6n89 October 28, 2025
Download PDF
Abstract: Traditional microscopes provide only 2D intensity information for transparent or unstained samples, limiting access to 3D morphology. Here, we present an in-line lensless digital holographic system using a 532 nm monochromatic light source. The unmodulated direct component serves as the reference wave, and the sample-diffracted component forms the interference, enabling single-frame 3D reconstruction via angular spectrum analysis. The system integrates a board-level CMOS camera with a simplified optical-mechanical structure, alongside automated preprocessing, conjugate suppression, and phase unwrapping. Unified sampling and coherence constraints yield a well-defined effective FOV, allowing quantitative 3D imaging of butterfly antennae and earthworm cross-sections. This low-cost, portable approach bypasses the need for expensive objectives and complex calibration, complementing—but not replacing—high-end instruments. Current limitations include axial calibration and phase noise control; future work will incorporate multi-frame phase shifting, learning-based conjugate suppression, and reflective imaging for strongly scattering samples. This platform provides accessible 3D imaging for entomology, biophotonics, biomimetic materials, and optical device research.
Keywords: Digital Holography; Interferometric Measurement; Three-Dimensional Imaging.
APA Citation: Yangxiao (Olivia) Ou (2025). Lens-Free Microscopic Imaging for 3D Analysis of Insect Microstructures and Ecological Applications. Transactions on Computer Science and Intelligent Systems Research, 11(1), 141-149. https://doi.org/10.62051/4esn6n89

References

  1. Gabor, D. “Holography, 1948-1971.” Science, vol. 177, no. 4046, 28 July 1972, pp. 299 – 313, science.sciencemag.org/content/177/4046/299, https://doi.org/10.1126/science.177.4046.299.
  2. Leith, Emmett N., and Juris Upatnieks. “Wavefront Reconstruction with Diffused Illumination and Three-Dimensional Objects*.” Journal of the Optical Society of America, vol. 54, no. 11, 1 Nov. 1964, p. 1295, https://doi.org/10.1364/josa.54.001295. Accessed 2 Mar. 2023.
  3. Colomb, Tristan, et al. "Advantages of digital holographic microscopy for real-time full field absolute phase imaging." Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XV. Vol. 6861. SPIE, 2008.
  4. Hayes-Rounds, Charity, et al. "Advantages of Fresnel biprism-based digital holographic microscopy in quantitative phase imaging." Journal of biomedical optics 25.8 (2020): 086501 - 086501.
  5. Zhang, Jialin, et al. “Resolution Analysis in a Lens-Free On-Chip Digital Holographic Microscope.” IEEE Transactions on Computational Imaging, vol. 6, Jan. 2020, pp. 697 – 710. https://doi.org/10.1109/tci.2020.2964247.
  6. Latychevskaia, Tatiana, et al. "Off-axis and inline electron holography: Experimental comparison." Ultramicroscopy 110.5 (2010): 472 - 482.
  7. Kim, Myung K. "Principles and techniques of digital holographic microscopy." SPIE reviews 1.1 (2010): 018005.
  8. Huang, Zhengzhong, and Liangcai Cao. “Quantitative Phase Imaging Based on Holography: Trends and New Perspectives.” Light Science & Applications, vol.13, no.1, June 2024, https://doi.org/10.1038/s41377 - 024 - 01453 - x.
  9. Guangjun, Wang, et al. “Comparison of Commonly Used Numerical Reconstruction Algorithms in Digital Holographic Microscopy.” Laser & Optoelectronics Progress, vol. 47, no. 3, Jan. 2010, p. 030901. https://doi.org/10.3788/lop47.030901.
  10. Zhang, Jiwei, et al. “A Review of Common-path Off-axis Digital Holography: Towards High Stable Optical Instrument Manufacturing.” Light Advanced Manufacturing, vol.2, no.3, Jan.2021, p.1. https://doi.org/10.37188/lam.2021.023.
  11. Kozacki, Tomasz, and Maksymilian Chlipala. "Color holographic display with white light LED source and single phase only SLM." Optics Express 24.3 (2016): 2189 - 2199.