Special Issue "Applications of Digital Holography in Biomedical Engineering"
I en specialutgåva av det vetenskapliga organet Applied Sciences kommer vi få läsa minst 4 nya HoloMonitorbaserade forskningsrapporter.Redan nu är 1 rapport offentlig.
Received: 24 December 2019 / Revised: 15 January 2020 / Accepted: 18 January 2020 / Published: 21 January 2020
Evaluation of the Impact of Imprinted Polymer Particles on Morphology and Motility of Breast Cancer Cells by Using Digital Holographic Cytometry
by
Megha Patel 1, 
Marek Feith 2, Birgit Janicke 3, Kersti Alm 3 and Zahra El-Schich 1,*
Megha Patel 1, Marek Feith 2, Birgit Janicke 3, Kersti Alm 3 and Zahra El-Schich 1,*
1
Department of Biomedical Sciences, Faculty of Health and Society, Malmö University, 205 06 Malmö, Sweden
2
Department of Physiology, Faculty of Medicine, Masaryk University, 625 00 Brno, Czech Republic
3
Phase Holographic Imaging AB, 223 63 Lund, Sweden
*
Author to whom correspondence should be addressed.
Abstract
Breast cancer is the second most common cancer type worldwide and breast
cancer metastasis accounts for the majority of breast cancer-related
deaths. Tumour cells produce increased levels of sialic acid (SA) that
terminates the monosaccharide on glycan chains of the glycosylated
proteins.
SA can contribute to cellular recognition, cancer invasiveness
and increase the metastatic potential of cancer cells. SA-templated
molecularly imprinted polymers (MIPs) have been proposed as promising
reporters for specific targeting of cancer cells when deployed in
nanoparticle format. The sialic acid-molecularly imprinted polymers
(SA-MIPs), which use SA for the generation of binding sites through
which the nanoparticles can target and stain breast cancer cells, opens
new strategies for efficient diagnostic tools. This study aims at
monitoring the effects of SA-MIPs on morphology and motility of the
epithelial type MCF-7 and the highly metastatic MDAMB231 breast cancer
cell lines, using digital holographic cytometry (DHC). DHC is a
label-free technique that is used in cell morphology studies of e.g.,
cell volume, area and thickness as well as in motility studies. Here, we
show that MCF-7 cells move slower than MDAMB231 cells. We also show
that SA-MIPs have an effect on cell morphology, motility and viability
of both cell lines. In conclusion, by using DH microscopy, we could
detect SA-MIPs impact on different breast cancer cells regarding
morphology and motility.
Keywords:
breast cancer; digital holographic cytometry; molecularly imprinted polymers; motility; sialic acid; viability
2.3.
DHC and Computer Software
For each cell line, 2 × 104 cells/well were seeded in a working volume of 1.8 mL in Sarstedt’s lumox®® multiwell 24 wells plate (Sarstedt, Inc. Nümbrecht, Germany). The cells were incubated for 24 h at 37 °C in humidified 5% CO2 atmosphere to allow to adhere. After incubation, 0.04 mg/mL of SA-MIPs were added to the wells.
Control wells were left without additive for both cell lines.
The plate was incubated for 1 h at 37 °C in humidified 5% CO2 atmosphere.
After the incubation, the standard lid was replaced with the HoloLidTM (Phase Holographic Imaging AB, Lund, Sweden).
The plate was mounted on a HoloMonitorTM M4 (Phase Holographic Imaging, AB), which is placed inside a cell incubator to ensure stable conditions of the cells during long-term experiments.
The HoloMonitorTM is equipped with a 635 nm low laser beam with 0.2 mW/cm2 to prevent phototoxicity effects on the cells.
The image analysis was performed with the proprietary AppSuite software (Phase Holographic Imaging AB). The images were acquired automatically every 30 min for 72 h.
3.2.
Different Motility between the Cell Lines Analysed with DHC
The motility analysis with DHC showed differences between MCF-7 and MDAMB231. The MDAMB231 cells had higher motility (Video S3) compared to the MCF-7 cells (Video S1) with up to twice the accumulated distance. The MDAMB231 cells incubated with SA-MIPs showed a slight decrease of the accumulated distance after 72 h, while for the MCF-7 cells the accumulated distance increased (Figure 2, Video S2–S4).
3.3.
Morphology Changes Detected with DHC MCF-7 and MDAMB231 cells (Figure 3B, D) treated with SA-MIPs for 72 hours had increased in thickness compared to untreated MCF-7 and MDAMB231 cells, respectively as detected by DHC (Figure 3A, C).
5. Conclusions
SA-MIPs affect motility, morphology and viability on both MCF-7 and MDAMB231 cell lines.
The MDAMB231 cells showed higher motility compared to the MCF-7 cells but with SA-MIPs treatment the motility decreased for the MDA-MB231 and increased for the MCF-7 cells.
The SAMIPs treatment over time affected cell morphology such as cell volume, area and thickness. In conclusion, DHC is a powerful tool to analyse the motility and morphology differences between cell lines as well as the SA-MIPs-induced cell response.
Min kommentar
2.3.
DHC and Computer Software
For each cell line, 2 × 104 cells/well were seeded in a working volume of 1.8 mL in Sarstedt’s lumox®® multiwell 24 wells plate (Sarstedt, Inc. Nümbrecht, Germany). The cells were incubated for 24 h at 37 °C in humidified 5% CO2 atmosphere to allow to adhere. After incubation, 0.04 mg/mL of SA-MIPs were added to the wells.
Control wells were left without additive for both cell lines.
The plate was incubated for 1 h at 37 °C in humidified 5% CO2 atmosphere.
After the incubation, the standard lid was replaced with the HoloLidTM (Phase Holographic Imaging AB, Lund, Sweden).
The plate was mounted on a HoloMonitorTM M4 (Phase Holographic Imaging, AB), which is placed inside a cell incubator to ensure stable conditions of the cells during long-term experiments.
The HoloMonitorTM is equipped with a 635 nm low laser beam with 0.2 mW/cm2 to prevent phototoxicity effects on the cells.
The image analysis was performed with the proprietary AppSuite software (Phase Holographic Imaging AB). The images were acquired automatically every 30 min for 72 h.
3.2.
Different Motility between the Cell Lines Analysed with DHC
The motility analysis with DHC showed differences between MCF-7 and MDAMB231. The MDAMB231 cells had higher motility (Video S3) compared to the MCF-7 cells (Video S1) with up to twice the accumulated distance. The MDAMB231 cells incubated with SA-MIPs showed a slight decrease of the accumulated distance after 72 h, while for the MCF-7 cells the accumulated distance increased (Figure 2, Video S2–S4).
3.3.
Morphology Changes Detected with DHC MCF-7 and MDAMB231 cells (Figure 3B, D) treated with SA-MIPs for 72 hours had increased in thickness compared to untreated MCF-7 and MDAMB231 cells, respectively as detected by DHC (Figure 3A, C).
 |
| Figure 3. 3D holograms presenting the untreated MCF-7 cells (A), MDAMB231 (C) and the treated MCF-7 cells (B), MDAMB231 (D) with 0.04 mg/mL of SA-MIPs after 72 h incubation. The colour scale represents the thickness and the scale bar represents 20 µm. |
SA-MIPs affect motility, morphology and viability on both MCF-7 and MDAMB231 cell lines.
The MDAMB231 cells showed higher motility compared to the MCF-7 cells but with SA-MIPs treatment the motility decreased for the MDA-MB231 and increased for the MCF-7 cells.
The SAMIPs treatment over time affected cell morphology such as cell volume, area and thickness. In conclusion, DHC is a powerful tool to analyse the motility and morphology differences between cell lines as well as the SA-MIPs-induced cell response.
Min kommentar
Denna studie är alltså från GlycoImaging-projektet och berättar om deras MIPSar (cancermarkörer) vid studier på 2 olika typer av bröstcancer,cellinjerna MCF-7 och MDAMB231 .
Man har använt uttrycket DHC (Digital Holographic Cytometry) istället för det vi i vanliga fall känner igen HoloMonitor som: DHM (Digital Holographic Microscope).
I Conclusions får vi info att deras cancermarkörer (MIPS) visar på avsedd effekt,att med deras MIP`s målsöka ämnet sialic acid (SA) som finns i bröstcancertumörer.Det ser man genom att använda HoloMonitor när man studerar cancertumörernas kännetecken.Vilket beskrivs i sista meningen : DHC is a powerful tool to analyse....
Vi får alltså besked att GlycoImaging projektet är framgångsrikt såtillvida att markörerna gör sitt jobb och fungerar som det var tänkt från början. Ett mycket bra besked !
Vi kan förmodligen snart börja spekulera kring Glycoprojektet och dess potential.
Övrigt
Man har använt uttrycket DHC (Digital Holographic Cytometry) istället för det vi i vanliga fall känner igen HoloMonitor som: DHM (Digital Holographic Microscope).
I Conclusions får vi info att deras cancermarkörer (MIPS) visar på avsedd effekt,att med deras MIP`s målsöka ämnet sialic acid (SA) som finns i bröstcancertumörer.Det ser man genom att använda HoloMonitor när man studerar cancertumörernas kännetecken.Vilket beskrivs i sista meningen : DHC is a powerful tool to analyse....
Vi får alltså besked att GlycoImaging projektet är framgångsrikt såtillvida att markörerna gör sitt jobb och fungerar som det var tänkt från början. Ett mycket bra besked !
Vi kan förmodligen snart börja spekulera kring Glycoprojektet och dess potential.
Övrigt
På länken hittar man kompletterande information som visar 4 filmer gjorda av HoloMonitor i studien.
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Det kan bli fler då sista inlämningsdatum är 15 Feb.
Planned Papers
The below list represents only planned manuscripts. Some of these manuscripts have not been received by the Editorial Office yet. Papers submitted to MDPI journals are subject to peer-review.
1. Anette Gjörloff Wingren
Email:anette.gjorloff-wingren@mau.se
Affiliation:Department of Biomedical Sciences, Faculty of Health and Society, Malmö University, Malmö, Sweden
2. Malin Lindstedt
Email:malin.lindstedt@immun.lth.se
Affiliation:Lunds universitet, Sweden
3. Robert Judson-Torres
Email:robert.judson-torres@hci.utah.edu
Affiliation:The University of Utah, USA
Email:anette.gjorloff-wingren@mau.se
Affiliation:Department of Biomedical Sciences, Faculty of Health and Society, Malmö University, Malmö, Sweden
2. Malin Lindstedt
Email:malin.lindstedt@immun.lth.se
Affiliation:Lunds universitet, Sweden
3. Robert Judson-Torres
Email:robert.judson-torres@hci.utah.edu
Affiliation:The University of Utah, USA
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