Biomod/2013/LMU/summary: Difference between revisions

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<s>In this section we want to give a short summary of the project. As we already were able to do some first steps into future experiments we will also give some details how the project will be continued.</s>
In this section we would like to present the most important aspects of our project once more and discuss the outcome of our work. Apart from this we would like to give an outlook on future projects based on our results beyond BIOMOD.
 
<span style="color:orange">In this section we would like to present the most important aspects of our project once more and discuss the outcome of our work. Apart from this we would like to give an outlook on future projects based on our results beyond BIOMOD.</span>


=Goals reached=
=Goals reached=
<s>The first question to answer if we want to qualify our developement is if the goals set at the beginning were reached. Due to the inhomogenity of the material and the foreseeable obstacles that come along with that we limited the goal of our project to a feasible extent. Namely we wanted to develope a proper functionalization of the nanodiamonds surface fulfilling three conditions:</s>
When assessing the project, it is essential to compare previously defined goals with the actual results. Inspired by the idea of developing a self-assembling single photon source, we decided to focus on one essential and so far unsolved problem as a feasible goal for one summer:<br />
The development of a method to functionalize the surface of nanodiamonds. Such a functionalization has to fulfill three conditions:<br />  
<br />  
<br />  
#It has to be biocompatible to allow a future in vivo use.<br />
#The functionalized nanodiamonds must offer surface groups which can be linked to specific docking sites of the DNA origami.<br />
#The functionalization should cause the nanodiamonds to be well dispersed, as single nanodiaomds should be attached to DNA origami.<br />
<br />  
<br />  
<span style="color:orange">When assessing the project, it is essential to compare previously defined goals with the actual results. Inspired by the idea of developing a self-assembling single photon source, we decided to focus on one essential and so far unsolved problem as a feasible goal for one summer:<br />
We were able to reach all of those aims:<br />
The development of a method to functionalize the surface of nanodiamonds. Such a functionalization has to fulfill three conditions:
</span>
 
 
#It has to be biocompatible to allow a future in vivo use.<br />
#The functionalized nanodiamonds must offer surface groups which can be linked to specific docking sites of the DNA-origami.<br />
#The functionalization should cause the nanodiamonds to be well dispersed, as single nanodiaomds should be attached to DNA-origami.<br />
 
<s>All this conditions are fulfilled by the developed coating. The included materials in the process are fully biocompatible and intended for the use at physiological conditions. In addition to that the coated diamonds dispersed well and thus could be bound specifically to our designed 6HB. This could be verified by TEM measurements. So we can report specific binding of functionalized nanodiamonds to DNA-Origami for the very first time.</s><br />


<span style="color:orange">We were able to reach all of those aims:<br />
All materials used are fully biocompatible and are intended for a use under physiological conditions. The nanodiamonds are well dispersed. Therefore they could be bound specifically to the 6HB structure we designed. Those result were verified by TEM measurements. We are happy to be able to report successful specific binding of functionalized nanodiamonds to DNA origami for the very first time.<br />
All materials used are fully biocompatible and are intended for a use under physiological conditions. The nanodiamonds are well dispersed. Therefore they could be bound specifically to the 6HB structure we designed. Those result were verified by TEM measurements. We are happy to be able to report successful specific binding of functionalized nanodiamonds to DNA-origami for the very first time.
[[Image:Emission excitation.png|thumb|400px|right|Normalized emission(red) and excitation(blue) spectra taken by FluoroLog]]
</span>
After providing the proof of principle of our approch, we already made some progress in improving the yield of our method. In the last samples we reached a yield of approximately 10% of structures with two diamonds while 50% showed at least one attachment.<br/>


Additionally we were already able to roughly determine the excitation and emission spectra of the used fluorescence diamonds. The excitation wavelength lies at 535nm, whereas the emission wavelength lies at 650nm. So the NVCs can be easily excited since commonly used laser systems such as doubled Nd:YAG work at a wavelength of 532nm. All this fluorescence measurements were taken by [[Biomod/2013/LMU/equipment#FluoroLog (Fluorescence spectrometer)|FluoroLog]].


<s>Our experiment was aimed to proof the principle but we are already made some progress in improving the yield of our method for use in further experiments.</s> <span style="color:orange">After providing the proof of principle of our approch, we are already made some progress in improving the yield of our method.</span>  Additionally we were already able to roughly determine the excitation and absortion spectra of the used fluorescence diamonds<span style="color:orange">( Link??? Image?? Proof???)</span>. This opens the ground for the next steps heading to realize the structure of Schietinger et al. (2009) in a self arranging structure.


=Next steps=
=Next steps=
[[Image:goldND.jpg|thumb|400px|right|Model of the self assembling nano diamond gold hybrid structure <span style="color:orange">Das andere Bild?</span>]]
[[Image:goldND.jpg|thumb|400px|right|Model of the self assembling nanodiamond gold hybrid structure]]
Since the initial motivation of the project was based on realizing diamond nanoparticles hybrid structures in a self assembling way we are already planning the next steps. <s>For this we will work together with the group of Prof. Högele which has the know-how and necessary equipment on disposal to carry out</s> <span style="color:orange"> To accomplish this we will work together with Prof. Högele’s group <span style="color:green">(Link)</span> which has the expertise and equipment available <s>which</s><span style="color:green">that</span> is necessary </span><span style="color:green">for</span> both detailed optical characterisation and single photon measurements at the nano-scale.<br />
Since the initial motivation of the project was based on realizing diamond nanoparticles hybrid structures in a self assembling way we are already planning the next steps. To accomplish this we will work together with [http://www.nano.physik.uni-muenchen.de/nanophotonics/home.html Prof. Högele’s group] which has the expertise and equipment available that is necessary for both detailed optical characterization and single photon measurements at the nano-scale.<br />


The long term objective is to synthesize a 6HB structure with nanodiamonds surrounded by goldnanoparticles on both sides. A visualization of <s>that</s> <span style="color:orange">the structure setup is shown </span>  <s>can be seen</s> on the picture to the right. <br />
The long term objective is to synthesize a 6HB structure with nanodiamonds surrounded by goldnanoparticles on both sides. A visualization of the structure setup is shown on the picture to the right. <br />
With this in mind we first tried to make some fluorescence pictures of the assembled nanodiamond 6HB structures. But the spectrum of the emission turned out to be a problem, since it is not in the measurement range of the fluorescence microscope available.
With this in mind we first tried to make some fluorescence pictures of the assembled nanodiamond 6HB structures. But the spectrum of the emission turned out to be a problem, since it is not in the measurement range of the fluorescence microscope available.
<s>Despite this due to the cooperation with Prof. Högele we are very confident to find a solution for that during the next weeks.</s> <span style="color:orange">However, we are confident to find a solution for this problem with the the cooperation of Prof. Högele.</span> After that we will be able to conduct the first single photon measurements which constitute the basis for the interpretation of the following experiments with goldnanoparticles-nanodiamonds arrangements.
However, we are confident to find a solution for this problem with the the cooperation of Prof. Högele. After that we will be able to conduct the first single photon measurements which constitute the basis for the interpretation of the following experiments with goldnanoparticles-nanodiamonds arrangements.
Regarding this we have already <s>done</s> <span style="color:orange">conducted</span>  some first experiments adding gold particles to <s>our</s> structures <span style="color:orange">developed by us.</span>  <s>but it turned out that</s> <span style="color:orange">Unfortunatly</span>  the goldparticles clustered with the nanodiamonds and thus inhibited the specific attachment to the 6HB.
Regarding this we have already conducted some first experiments adding gold particles to structures developed by us. Unfortunatly the goldparticles clustered with the nanodiamonds and thus inhibited the specific attachment to the 6HB.
But once we <s>will have jumped over</s> <span style="color:orange">overcome</span>  this last obstacle we will have the set up needed for a demonstration of the expected improvement of both excitation and emission rate of the NVC radiation as already demonstrated in the publication of Schietinger at al. of 2009.
But once we overcome this last obstacle we should be able to assemble the proposed nanodiamond gold hybrid structure similar to the one used in the paper of Schietinger et al. in a self assembling way.
If we can realize that it will be the first time to demonstrate that on a self arranged structure.


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In this section we would like to present the most important aspects of our project once more and discuss the outcome of our work. Apart from this we would like to give an outlook on future projects based on our results beyond BIOMOD.

Goals reached

When assessing the project, it is essential to compare previously defined goals with the actual results. Inspired by the idea of developing a self-assembling single photon source, we decided to focus on one essential and so far unsolved problem as a feasible goal for one summer:
The development of a method to functionalize the surface of nanodiamonds. Such a functionalization has to fulfill three conditions:

  1. It has to be biocompatible to allow a future in vivo use.
  2. The functionalized nanodiamonds must offer surface groups which can be linked to specific docking sites of the DNA origami.
  3. The functionalization should cause the nanodiamonds to be well dispersed, as single nanodiaomds should be attached to DNA origami.


We were able to reach all of those aims:

All materials used are fully biocompatible and are intended for a use under physiological conditions. The nanodiamonds are well dispersed. Therefore they could be bound specifically to the 6HB structure we designed. Those result were verified by TEM measurements. We are happy to be able to report successful specific binding of functionalized nanodiamonds to DNA origami for the very first time.

Normalized emission(red) and excitation(blue) spectra taken by FluoroLog

After providing the proof of principle of our approch, we already made some progress in improving the yield of our method. In the last samples we reached a yield of approximately 10% of structures with two diamonds while 50% showed at least one attachment.

Additionally we were already able to roughly determine the excitation and emission spectra of the used fluorescence diamonds. The excitation wavelength lies at 535nm, whereas the emission wavelength lies at 650nm. So the NVCs can be easily excited since commonly used laser systems such as doubled Nd:YAG work at a wavelength of 532nm. All this fluorescence measurements were taken by FluoroLog.


Next steps

Model of the self assembling nanodiamond gold hybrid structure

Since the initial motivation of the project was based on realizing diamond nanoparticles hybrid structures in a self assembling way we are already planning the next steps. To accomplish this we will work together with Prof. Högele’s group which has the expertise and equipment available that is necessary for both detailed optical characterization and single photon measurements at the nano-scale.

The long term objective is to synthesize a 6HB structure with nanodiamonds surrounded by goldnanoparticles on both sides. A visualization of the structure setup is shown on the picture to the right.
With this in mind we first tried to make some fluorescence pictures of the assembled nanodiamond 6HB structures. But the spectrum of the emission turned out to be a problem, since it is not in the measurement range of the fluorescence microscope available. However, we are confident to find a solution for this problem with the the cooperation of Prof. Högele. After that we will be able to conduct the first single photon measurements which constitute the basis for the interpretation of the following experiments with goldnanoparticles-nanodiamonds arrangements. Regarding this we have already conducted some first experiments adding gold particles to structures developed by us. Unfortunatly the goldparticles clustered with the nanodiamonds and thus inhibited the specific attachment to the 6HB. But once we overcome this last obstacle we should be able to assemble the proposed nanodiamond gold hybrid structure similar to the one used in the paper of Schietinger et al. in a self assembling way.