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Background
FITC (fluorescein isothiocyanate) is a fluorescent dye in biomedical studies and diagnosis. It's a yellow fluorescent light, excitation 490 nm and emission 520 nm, highly fluorescent and photostable. Because FITC is embedded in molecules with isothiocyanate groups, the chemical bonds it makes with the amino groups in molecules are permanent covalent bonds, marking target molecules. Further, FITC has differing fluorescence profiles with different pH conditions – the quantum yield is higher in alkaline conditions, and therefore its signal-to-noise ratio is improved in fluorescence microscopy imaging.
FITC (fluorescein isothiocyanate) is a fluorescent dye in biomedical studies and diagnosis. It's a yellow fluorescent light, excitation 490 nm and emission 520 nm, highly fluorescent and photostable. Because FITC is embedded in molecules with isothiocyanate groups, the chemical bonds it makes with the amino groups in molecules are permanent covalent bonds, marking target molecules. Further, FITC has differing fluorescence profiles with different pH conditions – the quantum yield is higher in alkaline conditions, and therefore its signal-to-noise ratio is improved in fluorescence microscopy imaging.
Figure 1. Schematic diagram of chemical synthesis of FITC-SiNPs (Source: Li J, et al. 2021)
Despite the broad application prospects of FITC in biomedical research, it faces several technical challenges. For instance, in certain situations, the fluorescence signal from FITC labels may be too intense, potentially interfering with experimental results. Additionally, the stability of FITC under varying pH levels and temperature conditions requires further optimization.
1. Li J, et al. Chemical conjugation of FITC to track silica nanoparticles in vivo and in vitro: An emerging method to assess the reproductive toxicity of industrial nanomaterials. Environ Int. 2021 Jul;152:106497.
2. Caprifico AE, et al. Biomedical and Pharmacological Uses of Fluorescein Isothiocyanate Chitosan-Based Nanocarriers. Macromol Biosci. 2021 Jan;21(1):e2000312.
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References
Colocalization of optical coherence tomography angiography with histology in the mouse retina
Optical coherence tomography angiography (OCT-A) allows in vivo, non-invasive, functional imaging of retinal perfusion. The purpose of this study was to determine the reliability of OCT-A in visualizing the complete retinal vasculature by comparing in vivo OCT-A images to matched ex vivo retinal tissue in mice. Adult female C57BL/6 mice were imaged to obtain OCT-A images of the superficial vascular complex, intermediate capillary plexus and deep capillary plexus. Z-stack fluorescence images of whole-mounted retinas, labeled for vascular endothelial cells by anti-isolectin immunohistochemistry and FITC-dextran perfusion, were generated. The OCT-A and fluorescence images were manually colocalized and vessel length measured for each of the techniques. Mean vessel length among all plexuses showed less than 13% difference between OCT-A and lectin immunohistochemistry and less than 4% difference between OCT-A and FITC-dextran perfusion. The strength of the correlation between OCT-A and lectin immunohistochemistry ranged from 0.46-0.95, while that between OCT-A and FITC-perfusion ranged from 0.67-0.88. OCT-A visualized retinal vasculature in vivo to a similar extent in matched ex vivo histology images. Our results show that OCT-A is a reliable method for acquiring in vivo images of retinal perfusion in mice, with the ability to differentiate each vascular plexus.
Long Non-coding RNA CCAT1 Sponges miR-454 to Promote Chemoresistance of Ovarian Cancer Cells to Cisplatin by Regulation of Surviving
Purpose Colon cancer-associated transcript 1 (CCAT1) was identified as an oncogenic long non-coding RNA (lncRNA) in a variety of cancers. However, there was a lack of understanding of the mechanism by which CCAT1 conferred cisplatin (also known as DDP) resistance in ovarian cancer cells. Materials and Methods Cell viability of A2780, SKOV3, A2780/DDP, and SKOV3/DDP cells upon cisplatin treatment was monitored by MTT assay. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) detected the expression levels of CCAT1 and miR-454. The effect of sh-CCAT1 on cisplatin response was investigated in xenografts study. Bioinformatic analysis, luciferase reporter assay and qRT-PCR were conducted to validate the direct interaction among CCAT1, miR-454, and survivin. Apoptosis was determined by flow cytometry after dual staining of Annexin-V-FITC/propidium iodide, and the expression of apoptosis-related proteins Bcl-2, Bax and survivin were detected by qRT-PCR and Western blotting. Xenograft study was conducted to monitor in vivo tumor formation. Results CCAT1 was highly expressed in cisplatin-resistant ovarian cancer cell line A2780/DDP and SKOV3/DDP. Knockdown of CCAT1 restored sensitivity to cisplatin in vitro and in vivo. Our data revealed that silencing of CCAT1 promoted cisplatin-induced apoptosis via modulating the expression of pro- or anti-apoptotic proteins Bax, Bcl-2, and survivin. CCAT1 directly interacted with miR-454, and miR-454 overexpression potentiated cisplatin-induced apoptosis. Survivin was identified as a functional target of miR-454, restoration of survivin attenuated the effect of miR-454 on cisplatin response. In addition, miR-454 inhibitor or overexpression of survivin was found to abolish sh-CCAT1-induced apoptosis upon cisplatin treatment. Conclusion CCAT1/miR-454/survivin axis conferred cisplatin resistance in ovarian cancer cells.
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Figure 1. Schematic diagram of chemical synthesis of FITC-SiNPs
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