journal article·2020-12-20Open access

Interferometric Measurement of TGF-β Induced Epithelial-Mesenchymal Transition of Tumor Cells

Rahmetullah Varol, Gokhan Bora Esmer, Huseyin Uvet

Applied Sciences· Volume 10· Issue 24

Abstract

A three-dimensional profile reconstruction of live cells in dynamic cell cultures is a challenging problem due to the highly scattering nature of cell mediums. Furthermore, it is an interesting problem since these cultures present the optimal in vitro conditions that most closely resemble the cells’ natural conditions. In this paper, we report a holographic method used for imaging during the process of treatment of dynamic cell cultures with transforming growth factor beta (TGF-β) and the subsequent epithelial-mesenchymal transition (EMT). The imaging of dynamic cell cultures presents many challenges for holographic techniques due to the highly scattering and high speed nature of the environment. Here we report the algorithmic workflow we used for decreasing the imaging noise due to the presence of cell medium and achieving high speed reconstruction rates in real time. We also report the prominent morphological features we extracted from the obtained depth maps throughout the experiment. We conducted experiments on four different cell lines: ONCO-DG1, HCT-116, MDA-MB-231, and HUVEC. We observed the EMT process throughout a 48 h period after treatment with TGF-β with 6 h intervals for each sample. We show some examples of the reconstructed depth maps of tumor cells during the EMT phase. From these depth maps we extract some morphological parameters and report how they change after the EMT process is completed. The obtained results indicate that the proposed method presents certain advantages from an optical perspective particularly for applications where a dynamical medium is present. These advantages are lower signal-to-noise ratio (SNR) values and a simple setup compared to the setups used in similar studies. Future studies on this application could lead to the development of a model for the EMT process and its effects on cell to cell interactions.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: Digital Holography and Microscopy · Microfluidic and Bio-sensing Technologies · Optical Coherence Tomography Applications

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Computer Engineering Department, Bogazici University, İstanbul 34342, Turkey

Electrical and Electronics Departments, Marmara University, İstanbul 34722, Turkey

Mechatronics Engineering Department, Yildiz Technical University, İstanbul 34349, Turkey

Funding

The Scientific and Technological Research Council of Turkey (TUBITAK)

References (21)

Show source-provided references
1. Phase contrast, a new method for the microscopic observation of transparent objects10.1016/S0031-8914(42)80035-X ↗
2. Digital holographic microscopy: A noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy10.1364/OL.30.000468 ↗
3. Autofocusing in digital holographic phase contrast microscopy on pure phase objects for live cell imaging10.1364/AO.47.00D176 ↗
4. Deep-tissue focal fluorescence imaging with digitally time-reversed ultrasound-encoded light10.1038/ncomms1925 ↗
5. Optical focusing deep inside dynamic scattering media with near-infrared time-reversed ultrasonically encoded (TRUE) light10.1038/ncomms6904 ↗
6. Engineering cancer microenvironments for in vitro 3-D tumor models10.1016/j.mattod.2015.05.002 ↗
7. Parameter-optimized digital holographic microscope for high-resolution living-cell analysis10.1364/AO.43.006536 ↗
8. Assessment of the flow velocity of blood cells in a microfluidic device using joint spectral and time domain optical coherence tomography10.1364/OE.21.024025 ↗
9. Cell shape identification using digital holographic microscopy10.1364/AO.54.007442 ↗
10. Quantitative assessment of cancer cell morphology and motility using telecentric digital holographic microscopy and machine learning10.1002/cyto.a.23316 ↗
11. Investigation of osteoblast cells behavior in polymeric 3D micropatterned scaffolds using digital holographic microscopy10.1364/AO.53.004850 ↗
12. Multispectral quantitative phase imaging of human red blood cells using inexpensive narrowband multicolor LEDs10.1364/AO.55.002521 ↗
13. Quantitative investigation on morphology and intracellular transport dynamics of migrating cells10.1364/AO.58.00G162 ↗
14. Hoang, V.T., Stȩpniewski, G., Czarnecka, K.H., Kasztelanic, R., Long, V.C., Xuan, K.D., Shao, L., Śmietana, M., and Buczyński, R. (2019). Optical Properties of Buffers and Cell Culture Media for Optofluidic and Sensing Applications. Appl. Sci., 9.10.3390/app9061145 ↗
15. Quantitative scoring of epithelial and mesenchymal qualities of cancer cells using machine learning and quantitative phase imaging10.1117/1.JBO.25.2.026002 ↗
16. Phase-shifting digital holography10.1364/OL.22.001268 ↗
17. TGF-β-induced epithelial to mesenchymal transition10.1038/cr.2009.5 ↗
18. Hao, Y., Baker, D., and ten Dijke, P. (2019). TGF-β-mediated epithelial-mesenchymal transition and cancer metastasis. Int. J. Mol. Sci., 20.10.3390/ijms20112767 ↗
19. Mechanisms of the epithelial-mesenchymal transition by TGF-β10.2217/fon.09.90 ↗
20. Improved watershed transform for medical image segmentation using prior information10.1109/TMI.2004.824224 ↗
21. Goodman, J.W. (2005). Introduction to Fourier Optics, Roberts and Company Publishers.