In this research, we developed a low-cost fingerprint nanoprobe based on surface-enhanced Raman scattering with favorable biocompatibility. Our probe enables label-free live cell bowel imaging and can detect two bacterial strings by providing molecular fingerprint information. We also conducted principal component analysis of single living cells.
Researchers have recently focused on improving sensitivity, reproducibility, and developing no source of trace and technique to provide reliable moleculary information so they can enable the submitting and detection of multiple analyze, and it can be combined with machine data analysis for improved performance. Obtaining specifically Raman signals from cells can be challenging due to the presence of many other chemicals in the sample, which can interfere with the background signal Compared with the frozen detection, the label-free source had actual high surface sensitivity and could realize the directional detection of space errors within cells. More importantly, it can provide the intrinsic chemistry of the in-out at a single cell level in an unrestricted manner under natural conditions.
In the future, our work will focus on the complete architecture of carbon dot-based system, including the nanostructure fabrication and the source vacuums. We also focus on the biomedical fabrication of this source system, such as cell research and cell-agnostic applications. Begin by adding 100 microliters of the prepared carbon dot solution into 200 microliters of chloroauric acid at room temperature for 10 seconds until a purple suspension is formed.
Next, centrifuge the purple suspension at 4, 000 G for 10 minutes at room temperature. Gently remove the supernatant using a pipette if removing the gold carbon dots composite nanoparticles colloids is difficult. Resuspend the gold carbon dots composite nanoparticles in 200 microliters of deionized water to wash away excess carbon dots.
Centrifuge the suspension once again, remove the supernatant, and obtain the core shell nanoparticles by redispersing them in 100 microliters of deionized water. Store the mixture at four degrees Celsius. After fabricating the gold carbon dots composite nanoparticles, place a sterile sapphire chip into the 24-well plates and seed cells with one milliliter of the medium into a single well.
Incubate the plates in a humid environment to attain 70 to 80%confluence. Add the gold carbon dots composite nanoparticles to the single well and incubate for four to six hours. At the end of the incubation, remove the medium from the sapphire chips.
Gently rinse the chips with PBS and dip the sapphire chips in PBS for SERS detection. To perform the SERS experiment, switch on the computer, Raman spectrometer, and 785 nanometer laser. Next, start the accompanying software and perform calibration using silicon wafers.
Click new measurement followed by spectral acquisition and adjust the spectrum range, then go to the acquisition tab, adjust the exposure time and laser power, and click okay to initiate a new spectral acquisition. Use a 20 times objective lens for a clear cellular image, then switch to a 50 times lens. After selecting a point on the cells for measurement, click run to initiate the process and then save the spectra.
To obtain SERS imaging of A549 cells, click new streamline image acquisition, select the range to be photographed, and click okay. Set the edge streamline to 785 nanometers, center to 1, 200 centimeter inverse, exposure duration to five seconds, and laser power to 50%The parameter can be modified as required. Click on new of live imaging, choose signal to baseline, and set the first limit to 725 and the second limit to 1, 700, then go to area setup, set the proper steps, and click on okay.
Finally, click run to start performing SERS imaging. SERS spectra revealed that, even when the concentration of methylene blue is as low as 10 to the minus nine molar, gold carbon dots composite nanoparticles exhibit excellent SERS performance compared to gold nanoparticles. SERS spectra acquired on 20 points exhibited high similarity.
One month after the placement of gold carbon dots composite nanoparticles at four degrees Celsius, the average SERS activity at 951, 185 and 1, 620 centimeters inverse showed a degree of decay of about 5.43%11.44%and 13.94%respectively. Gold carbon dots composite nanoparticles barely showed cytotoxicity to A549 cells. The mean spectrum of A549 cells is presented.
Gold carbon dots composite nanoparticle aggregates exhibit obvious SERS signals without noise background in the SERS mapping of A549 cells. Spectra from different cell points demonstrated the heterogeneity of components in various cytoplasmic regions.
