Label-Free Quantitative In Vitro Live Cell Imaging with Digital Holographic Microscopy

Kemper, Björn; Bauwens, Andreas; Bettenworth, Dominik; Götte, Martin; Greve, Burkhard; Kastl, Lena; Ketelhut, Steffi; Lenz, Philipp; Mues, Sarah; Schnekenburger, Jürgen; Vollmer, Angelika

doi:10.1007/11663_2019_6

Cited by 26 publications

(21 citation statements)

References 125 publications

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“…The retrieved refractive index values of RAW 264.7 cells (Fig. 4) are comparable to those published earlier (47, 48). The refractive indices of the other cell lines that were investigated in this study, to the best of our knowledge, have not been published before.…”

Section: Discussionsupporting

confidence: 87%

“…The refractive indices of the other cell lines that were investigated in this study, to the best of our knowledge, have not been published before. The comparison with data from other cell types published show that the refractive index values are within similar limits but dependent on the cell type (48). In order to use QPI tools and refractive index data as a biomedical marker, it is necessary to create a library of refractive indices for wide number of cell lines and their internal organelles.…”

Section: Discussionmentioning

confidence: 91%

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Refractive Index Changes of Cells and Cellular Compartments Upon Paraformaldehyde Fixation Acquired by Tomographic Phase Microscopy

et al. 2020

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Three-dimensional quantitative phase imaging is an emerging method, which provides the 3D distribution of the refractive index (RI) and the dry mass in live and fixed cells as well as in tissues. However, an insufficiently answered question is the influence of chemical cell fixation procedures on the results of RI reconstructions. Therefore, this work is devoted to systematic investigations on the RI in cellular organelles of live and fixed cells including nucleus, nucleolus, nucleoplasm, and cytoplasm. The research was carried out on four different cell lines using a common paraformaldehyde (PFA)-based fixation protocol. The selected cell types represent the diversity of mammalian cells and therefore the results presented provide a picture of fixation caused RI changes in a broader context. A commercial Tomocube HT-1S device was used for 3D RI acquisition. The changes in the RI values after the fixation process are detected in the reconstructed phase distributions and amount to the order of 10 −3. The RI values decrease and the observed RI changes are found to be different between various cell lines; however, all of them show the most significant loss in the nucleolus. In conclusion, our study demonstrates the evident need for standardized preparation procedures in phase tomographic measurements.

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Section: Discussionsupporting

confidence: 87%

Section: Discussionmentioning

confidence: 91%

Refractive Index Changes of Cells and Cellular Compartments Upon Paraformaldehyde Fixation Acquired by Tomographic Phase Microscopy

et al. 2020

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“…Other autofluorescence-based imaging methods, such as fluorescence lifetime imaging microscopy (FLIM) [46], have the advantage of subcellular resolution, allowing the monitoring of differentiation, redox state and metabolism of the investigated cell thus providing an excellent platform to characterize leukocytes [47]. Digital holographic microscopy through quantitative phase contrast imaging is another promising technique that has the ability to monitor multiple morphologic parameters and motility of the investigated cells [48] (see Fig. 1A).…”

Section: Technological Overview Of Label-free Biosensorsmentioning

confidence: 99%

Functional blood cell analysis by label-free biosensors and single-cell technologies

Szittner

Péter

Kurunczi

et al. 2022

Advances in Colloid and Interface Science

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“…Digital holographic microscopy (DHM) is an interferometry‐based variant of QPI that typically uses the classic holographic principle, with the difference that the hologram recording is performed by a digital image sensor using a coherent light source (16). Although a rather old methodology, DHM has taken off only recently due to the computational power for image reconstruction and robust “off‐axis” imaging tools replacing classical interferometer setups.…”

Section: Figurementioning

confidence: 99%

Morphology – Here I Come Again

Hayden

Klenk

2020

Cytometry Pt A

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WHAT is this good for," was the comment of our clinical cooperation partner, when we showed him the first imaging flow cytometry results using a microfluidic system and a customized quantitative phase microscope for high-throughput blood cell analysis. What went wrong? We have shown the hematology-oncology expert phase images of various leukocyte types. Our eyes were trained on these images showing round-shaped cells of different sizes with rather low-contrast but we could interpret the image quality and even differentiate leukocyte types. The clinician, however, looked at blood cell images he was never trained on and expected contrast similar to stained blood smear. Our first disappointment vanished when we showed him dot plots from image analysis, which pleased him as he started to become familiar due to his own expertise in flow cytometry. Showing him additional results on the discrimination potential of leukemia samples finally gave us a mark that we are looking at something with clinical relevance (1, 2). This meeting was a really good experience for the team working on phase imaging flow cytometry

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Label-Free Quantitative In Vitro Live Cell Imaging with Digital Holographic Microscopy

Cited by 26 publications

References 125 publications

Refractive Index Changes of Cells and Cellular Compartments Upon Paraformaldehyde Fixation Acquired by Tomographic Phase Microscopy

Refractive Index Changes of Cells and Cellular Compartments Upon Paraformaldehyde Fixation Acquired by Tomographic Phase Microscopy

Functional blood cell analysis by label-free biosensors and single-cell technologies

Morphology – Here I Come Again

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