Investigation of a hyperspectral Scanning Laser Optical Tomography setup for label-free cell identification.
Hyperspectral imaging
Scanning Laser Optical Tomography
Tomography
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
01 Aug 2024
01 Aug 2024
Historique:
received:
26
04
2024
accepted:
26
07
2024
medline:
2
8
2024
pubmed:
2
8
2024
entrez:
1
8
2024
Statut:
epublish
Résumé
The development of non-destructive, tomographic imaging systems is a current topic of research in biomedical technologies. One of these technologies is Scanning Laser Optical Tomography (SLOT), which features a highly modular setup with various contrast mechanisms. Extending this technology with new acquisition mechanisms allows us to investigate untreated and non-stained biological samples, leaving their natural biological physiology intact. To enhance the development of SLOT, we aimed to extend the density of information with a significant increase of acquisition channels. This should allow us to investigate samples with unknown emission spectra and even allow for label-fee cell identification. We developed and integrated a hyperspectral module into an existing SLOT system. The adaptations allow for the acquisition of three-dimensional datasets containing a highly increased information density. For validation, artificial test objects were made from fluorescent acrylic and acquired with the new hyperspectral setup. In addition, measurements were made on two different human cell spheroids with an unknown spectra, to test the possibilities of label-free cell identification. The validation measurements of the artificial test target show the expected results. Furthermore, the measurements of the biological cell spheroids show small variations in their tomographic spectrum that allow for label-free cell type differentiation. The results of the biological sample demonstrate the potential of label-free cell identification of the newly developed setup.
Identifiants
pubmed: 39090238
doi: 10.1038/s41598-024-68685-0
pii: 10.1038/s41598-024-68685-0
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
17861Subventions
Organisme : European Regional Development Fund and the state of Lower Saxony
ID : 85031498
Informations de copyright
© 2024. The Author(s).
Références
Röntgen, W. C. Ueber eine neue Art von Strahlen. Ann. Phys. 300, 1–11 (1898).
doi: 10.1002/andp.18983000102
Buzug, T. Computed tomography: From photon statistics to modern cone-beam CT. Comput. Tomogr. From Phot. Stat. to Mod. Cone-Beam CT 1–521 (2008).
Schmitt, R. M. Ultrasound Computed Tomography: From the Past to the Future. 25–35 (2002).
Fujimoto, J. G. Optical coherence tomography: Technology and applications. In 2002 IEEE/LEOS Int. Conf. Opt. MEMs, OMEMS 2002-Conf. Dig. 147–148 (2002).
Pawley, J. B. Handbook of biological confocal microscopy: Third edition. In Handb. Biol. Confocal Microsc. 3rd ed, 1–985 (2006).
Lorbeer, R.-A. et al. Highly efficient 3D fluorescence microscopy with a scanning laser optical tomograph. Opt. Express 19, 5419 (2011).
doi: 10.1364/OE.19.005419
Sharpe, J. et al. Optical projection tomography as a tool for 3D microscopy and gene expression studies. Science 296, 541–545 (2002).
doi: 10.1126/science.1068206
Nolte, L. et al. Enabling second harmonic generation as a contrast mechanism for optical projection tomography (OPT) and scanning laser optical tomography (SLOT). Biomed. Opt. Express 9, 2627 (2018).
doi: 10.1364/BOE.9.002627
Nolte, L. et al. Scanning laser optical tomography for in toto imaging of the murine cochlea. PLoS ONE 12, 1–10 (2017).
doi: 10.1371/journal.pone.0175431
Arai, Y., Yamamoto, T., Minamikawa, T., Takamatsu, T. & Nagai, T. Spectral fingerprinting of individual cells visualized by cavity-reflection-enhanced light-absorption microscopy. PLoS ONE 10, 1–13 (2015).
doi: 10.1371/journal.pone.0125733
Stelzle, F. et al. Tissue discrimination by uncorrected autofluorescence spectra: A proof-of-principle study for tissue-specific laser surgery. Sensors 13, 13717–13731 (2013).
doi: 10.3390/s131013717
Benecke, H., Johannsmeier, S., May, T. & Ripken, T. Enabling hyperspectral acquisition for scanning laser optical tomography. In BiOS, 17 (2022).
Hohenhoff, G. et al. Comparison of SLOT and μ-CT investigation of 3D printed polymer parts for quality assurance. J. Laser Appl. 32, 022051 (2020).
doi: 10.2351/7.0000084
Heidrich, M. et al. 3D imaging of biofilms on implants by detection of scattered light with a scanning laser optical tomograph. Biomed. Opt. Express 2, 2982 (2011).
doi: 10.1364/BOE.2.002982
Kellner, M. et al. Imaging of the mouse lung with scanning laser optical tomography (SLOT). J. Appl. Physiol. 113, 975–983 (2012).
doi: 10.1152/japplphysiol.00026.2012
Kamin, H. et al. Correlative imaging and quantification of the tumor microenvironment of triple-negative-breast-cancer using lightsheet microscopy, scanning laser optical tomography, and TPEF (Conference Presentation), Vol. 14, 4 (2023).
Kellner, M. et al. A combined method for correlative 3D imaging of biological samples from macro to nano scale. Sci. Rep. 6, 1–12 (2016).
doi: 10.1038/srep35606
Kremer, J. R., Mastronarde, D. N. & McIntosh, J. R. Computer visualization of three-dimensional image data using IMOD. J. Struct. Biol. 116, 71–76 (1996).
doi: 10.1006/jsbi.1996.0013
Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).
doi: 10.1038/nmeth.2019
Kubitscheck, U. Fluorescence Microscopy: From Principles to Biological Applications (Wiley, 2013).
doi: 10.1002/9783527671595
Greivenkamp, J. E. Field Guide to Geometrical Optics (SPIE, 2004).
doi: 10.1117/3.547461
Lipps, C. et al. Expansion of functional personalized cells with specific transgene combinations. Nat. Commun. 9, 1–12 (2018).
doi: 10.1038/s41467-018-03408-4
Chen, L. et al. Hyperspectral scanning laser optical tomography. J. Biophotonics 12, 1–9 (2019).
doi: 10.1002/jbio.201800221
Gunaratne, R. et al. Machine learning classification of human joint tissue from diffuse reflectance spectroscopy data. Biomed. Opt. Express 10, 3889 (2019).
doi: 10.1364/BOE.10.003889