Synthesis of Fluorescents AuNCs

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Hi there. I present the approach of the synthesis of AuNC with the aim that they have the fluorescence property, i am looking for some help to identify failures in my procedure and achieve success in obtaining the product. Thanks!
Hi everyone, as a degree project to obtain a chemistry degree, i am developing the Synthesis of Fluorescent Gold Nanoclusters (AuNC). I have investigated the route to obtain nanoestructures of nanometric size less than five nanometers, reaction conditions, and so far i have managed to obtain a reaction product with no surface plasmon band and in the DLS analysis i have managed to show the size less than 5 nm. However, the fluorescence spectroscopy characterization doesn't give me any fluorescence emission results from my AuNCs. So, i'm going to share with you the synthesis route that i have proposed and other details, so that you please help me identify errors or omission of steps and possible causes of why i don't obtain fluorescent NCs.

As a bingind ligand i am using synthetic peptides, the peptide sequence that i am using for the tests is: Ac-CTKKCGLWSILKGVGKI-NH2
The Au-Pep ratios is 1:3
As a gold precursor i am using HAuCl4
The solvent is water
The reducing agent is Ascorbic Acid
To increase the reaction to alkaline pH i am using NaOH

Synthesis:

The first step is to increase the pH of the peptide to generate the availability of its negative charges, then to a 3mM solution of peptide i add NaOH up to pH 12 and i take it to the ultrasonic bath equipment for 10 minutes. After 10 minutes i add the 1mM gold solution and a little amount of ascorbic acid 50mM, then 10 minutes in the ultrasonic bath equipment and finally i pass it to the agitation equipment for 24h at 37ºC.

After this procedure, i run a test with a 365 nm lamp and the product fluoresces, but when i take it to the molecular fluorescence equipment i get no emission result and i don't realize what i'm omitting in order no to get a product that fluoresces. :c

Plese help me!

Thank you so much!

Note: The reducing agent that i proposed is with the aim of not using a strong, toxic and polluting agent. The peptide is provide by the university and i cannot change it. Please, let me know if you need more details to understand what's going on with this procedure.
 
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What excitation frequency are you using for the molecular fluorescence equipment?
 

Related to Synthesis of Fluorescents AuNCs

What is the synthesis of fluorescent AuNCs?

The synthesis of fluorescent AuNCs involves the reduction of gold salts in the presence of stabilizing ligands to produce small gold nanoclusters with unique fluorescent properties.

What are the applications of fluorescent AuNCs?

Fluorescent AuNCs have a wide range of applications in bioimaging, sensing, drug delivery, and catalysis due to their excellent photostability, biocompatibility, and tunable emission wavelengths.

How are fluorescent AuNCs characterized?

Fluorescent AuNCs are typically characterized using techniques such as UV-Vis spectroscopy, fluorescence spectroscopy, transmission electron microscopy (TEM), and dynamic light scattering (DLS) to determine their size, shape, stability, and optical properties.

What factors influence the synthesis of fluorescent AuNCs?

The synthesis of fluorescent AuNCs can be influenced by factors such as the choice of reducing agent, stabilizing ligands, reaction temperature, pH, and reaction time, which can all impact the size, shape, and fluorescence properties of the nanoclusters.

Are there any challenges in the synthesis of fluorescent AuNCs?

Some challenges in the synthesis of fluorescent AuNCs include controlling the size and monodispersity of the nanoclusters, optimizing the fluorescence quantum yield, and ensuring the stability of the nanoclusters in various biological and environmental conditions.

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