Nanoparticle Uptake in the Aging and Oncogenic Drosophila Midgut Measured with Surface-Enhanced Raman Spectroscopy
<p>SERS intensity at various nanoparticle relative concentrations. (<b>a</b>) The SERS nanoparticles demonstrate a characteristic spectrum that diminishes as they are serially diluted. The characteristic peaks appearing at 1203, 520 and 557 cm<sup>−1</sup> are highlighted. (<b>b</b>) A regression model was applied to the intensity of the 1203 cm<sup>−1</sup> peak, showing a logarithmic response of the nanoparticles as a function of concentration.</p> "> Figure 2
<p>Examples of (<b>a</b>) OR and (<b>b</b>) <span class="html-italic">w<sup>1118</sup></span> Smurf females with light diffused blue color (<b>left</b>) and non-Smurf (<b>right</b>).</p> "> Figure 3
<p>SERS NP uptake by young and old females. (<b>a</b>) Raman spectra from young and old flies, <span class="html-italic">w<sup>1118</sup></span> and OR, with and without clearance. The spectra demonstrate many Raman peaks intrinsic to the fly body. Spectra of individual flies are shown in grey and the group average in blue. The shaded bands indicate the areas of the peaks specific to the SERS NPs. (<b>b</b>) Regression (nn-LS) scores indicate moderate SERS signals in the flies fed with SERS NPs, with no statistically significant differences, as indicated by the <span class="html-italic">p</span>-values. Dots are individual fly measurements, the bars show the mean, and the whiskers the standard deviation. For each condition, 5 to 7 flies were scanned.</p> "> Figure 4
<p>SERS NP uptake by mutant flies. (<b>a</b>) Raman spectra for esg<sup>ts</sup>-Ras<sup>V12</sup> flies on day 2, 4 and 6 of induction of Ras<sup>V12</sup> oncogene in their midgut progenitors. Spectra of individual flies are shown in grey and the group average in blue. The shaded bands indicate the areas of the peaks specific to the SERS NPs. (<b>b</b>) Mean nn-LS scores and <span class="html-italic">p</span>-values as calculated for the different groups, show a small but statistically significant uptake for the 2-day condition, which decreases with disease progression. For each experimental condition, 10 flies were scanned.</p> "> Figure 5
<p>Raman maps of excised fly midguts. (<b>a</b>) Photograph of excised guts for NP-fed <span class="html-italic">esg<sup>ts</sup>-Ras<sup>V12</sup></span> flies. The superimposed colormap represents the nn-LS scores obtained from the different areas. Scale bar: 10 mm. (<b>b</b>) Detailed view from the sample areas indicated by boxes in (<b>a</b>). The optical microscopy images are shown with and without the colormap superimposed, for comparison. The areas with high nn-LS scores in their posterior midgut region display a reddish metallic texture, indicative of the presence of SERS NPs, particularly evident in (<b>bii</b>). Scale bar: 1 mm. (<b>c</b>) Representative Raman spectra corresponding to the pixels from panel (<b>b</b>) indicated with red crosses (×). Area (<b>bii</b>) features an exceptionally bright signal, whereas other areas display the characteristic peaks at lower intensities. (<b>d</b>–<b>f</b>) Similar to panels (<b>a</b>–<b>c</b>) but from control flies not fed with nanoparticles. Regression signals and Raman spectra have no indication of SERS nanoparticles, as expected.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
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- Fly production
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- SERS NP synthesis
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- Administration of SERS nanoparticles
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- Whole-fly Raman measurements
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- Midgut preparation for Raman imaging
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- Smurf assay for gut leakiness
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- Statistical analysis
3. Results
3.1. SERS Nanoparticle Characterization
3.2. Nanoparticle Uptake by the Gut of Young and Old Flies
3.3. Nanoparticle Uptake by Oncogenic Flies
3.3.1. Whole-Abdomen Measurements
3.3.2. Excised Gut Measurements
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Strain | OR | w1118 | ||||||
---|---|---|---|---|---|---|---|---|
Condition | Young | Aged | Young | Aged | ||||
n | % | n | % | n | % | n | % | |
Total | 63 | 100.0 | 17 | 100.0 | 42 | 100.0 | 24 | 100.0 |
Dead | 2 | 3.2 | 1 | 5.9 | 2 | 4.8 | 3 | 12.5 |
Non-Smurf | 58 | 92.1 | 9 | 52.9 | 34 | 81.0 | 11 | 45.8 |
Smurf | 3 | 4.8 | 7 | 41.2 | 6 | 14.3 | 10 | 41.7 |
p-value | 0.0004 | 0.0124 |
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Christou, M.; Fidelix, A.; Apidianakis, Y.; Andreou, C. Nanoparticle Uptake in the Aging and Oncogenic Drosophila Midgut Measured with Surface-Enhanced Raman Spectroscopy. Cells 2024, 13, 1344. https://doi.org/10.3390/cells13161344
Christou M, Fidelix A, Apidianakis Y, Andreou C. Nanoparticle Uptake in the Aging and Oncogenic Drosophila Midgut Measured with Surface-Enhanced Raman Spectroscopy. Cells. 2024; 13(16):1344. https://doi.org/10.3390/cells13161344
Chicago/Turabian StyleChristou, Maria, Ayobami Fidelix, Yiorgos Apidianakis, and Chrysafis Andreou. 2024. "Nanoparticle Uptake in the Aging and Oncogenic Drosophila Midgut Measured with Surface-Enhanced Raman Spectroscopy" Cells 13, no. 16: 1344. https://doi.org/10.3390/cells13161344