Lepton Anyone who think a science that specifically deals with macromolecules isn't real science should be put in jail or stoned at the city gates... I only noticed your comment because that Smock guy commented
@George92ut Thank you for your comment. Sorry for the confusion. The angle axis in the plots are defined as relative to the NORMAL of the chip instead of the angle label shown at 0:43 seconds.
The shape and location of the SPR dip can then be used to convey information about the sensor surface. SPR technology is commonly utilized for the study of molecular binding interactions between free anolyte molecules in solution and probe molecules which are linked to or immobilized onto the sensor surface. As a molecular binding event takes place the angular position of this dark band shifts, and a shift in the reflectivity curve can also be observed.
Surface plasmon resonance, or SPR, has emerged as a powerful optical detection technique for studying label free bio-molecular interactions in real-time within a variety of diverse applications such as life science, electrochemistry, chemical vapor detection, food and environmental safety and beyond.
This is a direct method of detection which avoids the drawbacks of labels. Another powerful aspect of SPR technology is its ability to observe the time dependent binding interaction between molecules, by monitoring the change in SPR response over time, researchers can study the kinetics of molecular binding events.
Due to its simplicity, nearly every commercially available SPR instrument uses a detection scheme called the Kretschmann Configuration, in this set up a light source passes through a prism, reflects off the backside of the sensor chip surface and into a detector. At a certain incident angle known as the resonance angle, light is absorbed by the electrons in the metal film of the sensor chip causing them to resonate.
These resonating electrons are also known as surface plasmons which are sensitive to their surrounding environment. The result is an intensity loss in the reflected beam which appears as a dark band and can be seen as a dip in the SPR Reflection Intensity Curve.
While sample is continuously delivered to the sensor surface anolyte molecules continue to bind and the number of available binding sites decreases, corresponding to a decrease in binding rate. As the SPR response levels off the system approaches equilibrium and the number of molecules binding and unbinding become equal. When no more anolyte is introduced to the system, the molecules will continue to unbind resulting in a decrease in SPR response.
. In the first example, an anolyte with a low affinity to the immobilized ligand on the sensor surface is introduced and therefore negligible binding interaction is observed. However, when an anolyte with higher affinity to the immobilized ligand is introduced into the system, then binding events are more readily observed. Initially there are many binding sites available resulting in a rapid increase in SPR response that occurs as anolyte begins to bind to the ligand.
. The information that can be obtained from SPR serves as a simple precise and label free technique for quantitative analysis of molecular binding interactions.
you question is answered in two parts. second part: Once the sample is removed, its observed contribution due to binding is removed. This results in a decreasing SPR response. When the unbinding process is complete, then the system will reach steady state.
Good question. During the sample's exposure to the surface, the sample molecules will continually bind and unbind with immobilized ligands on the sensor surface. This is a very dynamic process that occurs due to the nature of affinity binding interactions. Though the kinetic rates for association and dissociation may differ from each other, eventually the amount of sample binding and unbinding will reach equilibrium/steady state for a giving concentration.(i.e. Langmuir isotherm)
. The association constant, kA, can be extracted from the behavior of the binding response and likewise the dissociation constant, kD, can be extracted from the unbinding response. The ratio of these two rate constants can yield the binding affinity of the system.
Hello! Thanks for the good video. I have one question regarding the SPR response: what causes the response? is it the change of SPR angle with a fixed angle detector or the increased light absorbance caused by a higher concentration of analyte in the range of the evanescent wave?
It’s index of refraction change just above coverslip, so loss of signal due to increased scattering, since that also takes into account change in reflection angle. So yes, simple mass scatters light effect. I’m not sure how sensitive it is to change - one would think the layer thickness must need to change on the order of the size of the photons- so 200-400 nm, which would be tens of proteins deep, which is nonsensical. That’s the beauty of spr - the gold surface greatly reduces the size scale of photons to one or more layers of proteins, in effect! After that you just need a good detector for measuring small light changes in large signal which we have. I presume the light is exciting along the path of its resonance intensity minimum, and then see increase in emission intensity.
But, isn't the shift a binary indicator? It speaks to presence or absence of Target analyte. What about more analog sensing? Like concentration of analyte?
What software did you use to make this video?? I want to make something like this for my work but not able to understand what software to start with. Please help in this regards. Thank you,
Worth more than 10 papers on the argument. Is there any chance to have a little comment on how the oscillating electrons feel the surrounding? And by the way if I well understood the method the gold film has to be VERY think right? Either way the electrons on the other face of the gold film couldn't feel the effect of the incident radiaton right?
Thanks for the video, I finally understand whats is the "dip". I have a question, It is posibibly to detectc an analyte in a sensor chip if this isn't funcionalited? I mean, when it's only the gold layer
SPR based devices detects local changes at or near the sensor surface by measuring changes in refractive index vs. time. These refractive index changes are plotted in SPR response plots, which are also called sensorgrams. The sensorgrams are used to derive important information about how an analyte interacts with the sensor surface, i.e. binding interaction selectivity and rates. In the case of using a bare gold sensor, if the analyte nears or binds to the gold surface, this would be observed in the sensorgram as an increase in SPR response. By studying the rate of SPR signal response change at different analyte concentrations, quantitative analyte interaction information can be extracted such to what extent an analyte is simply passing by, truly binding, non-specifically binding, or physically absorbing. hope this helps.
Thank you so much!! It really helped me. Then, even if the sensor is pure gold, the sensorgram takes the same form than a functionalized sensor (base line, association, dissociation, regeneration and baseline again) of course it depends for the concentration and the analyte itself. I'm asking because i used to think that a sensorgram produced with a chip non-functionalized looked diferent than a sensorgram with a functionalized sensor.
Brilliant video, thanks! However just to note that at ~1:33 the animation shows the intensity shifting to a lower angle, whilst the corresponding graph shows it shifting to a higher angle. The angle I'm going on is the one you defined at ~0:43!
This is a nice video, but I think the narration is a little too fast. A few more well-timed pauses would help ideas sink in. Thank you for posting this video though.
Explained it better than my Molecular Biology prof, thanks.
Huh?
Lepton Excuse me? Where do you live? I'll fight you right now--- it's 100% real science.
Lepton Anyone who think a science that specifically deals with macromolecules isn't real science should be put in jail or stoned at the city gates... I only noticed your comment because that Smock guy commented
Lepton which city do you currently reside in?
Lepton I've a guess that I'm talking to an American. You people give off a cyber scent
bless y'all for this vid. My school's chem dept. thinks its 100% fine to just point us to a textbook for this
This was way more clear than my professor!
@George92ut
Thank you for your comment. Sorry for the confusion. The angle axis in the plots are defined as relative to the NORMAL of the chip instead of the angle label shown at 0:43 seconds.
Thank you for this simple & on-point explanation.
The shape and location of the SPR dip can then be used to convey information about the sensor surface. SPR technology is commonly utilized for the study of molecular binding interactions between free anolyte molecules in solution and probe molecules which are linked to or immobilized onto the sensor surface. As a molecular binding event takes place the angular position of this dark band shifts, and a shift in the reflectivity curve can also be observed.
Surface plasmon resonance, or SPR, has emerged as a powerful optical detection technique for studying label free bio-molecular interactions in real-time within a variety of diverse applications such as life science, electrochemistry, chemical vapor detection, food and environmental safety and beyond.
This is a direct method of detection which avoids the drawbacks of labels. Another powerful aspect of SPR technology is its ability to observe the time dependent binding interaction between molecules, by monitoring the change in SPR response over time, researchers can study the kinetics of molecular binding events.
Thanks this was a very clear but still easy to understand explanation video. Really helped me in understanding my lab.
hey fellow once... How you doing?
I was pretty lost until this video. Thanks for the simple explanation!
What a simple yet effective presentation!
Due to its simplicity, nearly every commercially available SPR instrument uses a detection scheme called the Kretschmann Configuration, in this set up a light source passes through a prism, reflects off the backside of the sensor chip surface and into a detector. At a certain incident angle known as the resonance angle, light is absorbed by the electrons in the metal film of the sensor chip causing them to resonate.
Thought this would take a while to understand but the video did it in less than 5 minutes. Thank you so much!
Fantastic presentation, very clear.
i like ur speaking.its really clear and easy to understand for non english speaker like me.thx u
These resonating electrons are also known as surface plasmons which are sensitive to their surrounding environment. The result is an intensity loss in the reflected beam which appears as a dark band and can be seen as a dip in the SPR Reflection Intensity Curve.
Nicely presented, clearly explained. Well done!
While sample is continuously delivered to the sensor surface anolyte molecules continue to bind and the number of available binding sites decreases, corresponding to a decrease in binding rate. As the SPR response levels off the system approaches equilibrium and the number of molecules binding and unbinding become equal. When no more anolyte is introduced to the system, the molecules will continue to unbind resulting in a decrease in SPR response.
. In the first example, an anolyte with a low affinity to the immobilized ligand on the sensor surface is introduced and therefore negligible binding interaction is observed. However, when an anolyte with higher affinity to the immobilized ligand is introduced into the system, then binding events are more readily observed. Initially there are many binding sites available resulting in a rapid increase in SPR response that occurs as anolyte begins to bind to the ligand.
. The information that can be obtained from SPR serves as a simple precise and label free technique for quantitative analysis of molecular binding interactions.
This was super helpful and insightful! Thanks!
you question is answered in two parts. second part:
Once the sample is removed, its observed contribution due to binding is removed. This results in a decreasing SPR response. When the unbinding process is complete, then the system will reach steady state.
?
Thank you so much! This made it so much clearer
Good question. During the sample's exposure to the surface, the sample molecules will continually bind and unbind with immobilized ligands on the sensor surface. This is a very dynamic process that occurs due to the nature of affinity binding interactions. Though the kinetic rates for association and dissociation may differ from each other, eventually the amount of sample binding and unbinding will reach equilibrium/steady state for a giving concentration.(i.e. Langmuir isotherm)
Dear everyone credited with building this video,
I love you all.
-Jay
Thanks it is quite good , I didn`t get a good grasp of it till I went to wikipedia first.
good explanation and animation
Brilliant video. You have the thanks of a seriously confused biomedical scientist
What an insightful video! Thanks!
V. good and simple explanation. Thankyou!
. The association constant, kA, can be extracted from the behavior of the binding response and likewise the dissociation constant, kD, can be extracted from the unbinding response. The ratio of these two rate constants can yield the binding affinity of the system.
Greatly explained. its cleared everything
Beautiful voice, love the accent!
yes, the voice actually arouses me positively
sorcery!! explained so clearly. great job.
Thank you for your work!
Great explanation, thanks!
Explicit instruction! Thank you!
Hello!
Thanks for the good video. I have one question regarding the SPR response:
what causes the response? is it the change of SPR angle with a fixed angle detector or the increased light absorbance caused by a higher concentration of analyte in the range of the evanescent wave?
It’s index of refraction change just above coverslip, so loss of signal due to increased scattering, since that also takes into account change in reflection angle. So yes, simple mass scatters light effect. I’m not sure how sensitive it is to change - one would think the layer thickness must need to change on the order of the size of the photons- so 200-400 nm, which would be tens of proteins deep, which is nonsensical. That’s the beauty of spr - the gold surface greatly reduces the size scale of photons to one or more layers of proteins, in effect! After that you just need a good detector for measuring small light changes in large signal which we have. I presume the light is exciting along the path of its resonance intensity minimum, and then see increase in emission intensity.
Nice explanation!
But, isn't the shift a binary indicator? It speaks to presence or absence of Target analyte. What about more analog sensing? Like concentration of analyte?
Thanks. Nice explanation of binding kinetics
fantastic video, thank you
can you please tell me which software use for surface plasmon resonance Implementation
Very helpful explanation - thank you.
Was very helpful, thank you
Awesome explanation
Great video, thanks a lot!
Thank you for sharing this video
What is the reason of using prism ?
awesome video..thanks alot..can u you tell me please which animation software you used in this video?
very nicely explained.
Helped so much thank you!
Fantastic.
What software did you use to make this video?? I want to make something like this for my work but not able to understand what software to start with. Please help in this regards. Thank you,
Worth more than 10 papers on the argument. Is there any chance to have a little comment on how the oscillating electrons feel the surrounding? And by the way if I well understood the method the gold film has to be VERY think right? Either way the electrons on the other face of the gold film couldn't feel the effect of the incident radiaton right?
www.in4tex.com/plasmon-spectrometer-resonance/
Thanks for the video, I finally understand whats is the "dip". I have a question, It is posibibly to detectc an analyte in a sensor chip if this isn't funcionalited? I mean, when it's only the gold layer
SPR based devices detects local changes at or near the sensor surface by measuring changes in refractive index vs. time. These refractive index changes are plotted in SPR response plots, which are also called sensorgrams. The sensorgrams are used to derive important information about how an analyte interacts with the sensor surface, i.e. binding interaction selectivity and rates. In the case of using a bare gold sensor, if the analyte nears or binds to the gold surface, this would be observed in the sensorgram as an increase in SPR response. By studying the rate of SPR signal response change at different analyte concentrations, quantitative analyte interaction information can be extracted such to what extent an analyte is simply passing by, truly binding, non-specifically binding, or physically absorbing. hope this helps.
Thank you so much!! It really helped me. Then, even if the sensor is pure gold, the sensorgram takes the same form than a functionalized sensor (base line, association, dissociation, regeneration and baseline again) of course it depends for the concentration and the analyte itself. I'm asking because i used to think that a sensorgram produced with a chip non-functionalized looked diferent than a sensorgram with a functionalized sensor.
Super helpful. I get it now.
SPR for dummies. Thank you :)
very nice video, thanks for uploading..
Well, wish I could have back the hr I spent reading wikipedia and various unintelligible resources and just would have watched this first.
awesome video
Thank you
Brilliant video, thanks! However just to note that at ~1:33 the animation shows the intensity shifting to a lower angle, whilst the corresponding graph shows it shifting to a higher angle. The angle I'm going on is the one you defined at ~0:43!
Very nice
Fantastic
THANK YOU!
Thanks very GOOD
Awesome thank you if you go below 0.08nm then your entering the afterlife spectrum ;)
SPR explained, indeed!
This is a nice video, but I think the narration is a little too fast. A few more well-timed pauses would help ideas sink in. Thank you for posting this video though.
She sounds like being drunk when running on 0.75x speed.
0:27
It would be more helpful if the narration was a bit slow!
the first surface plasmon resonance microscope:
www.in4tex.com/surface-plasmon-resonance-microscopy/
میکروسکوپ تشدید پلاسمون سطحی
Liked it
THANKS!!!!!!!!!!!!!!!!!!!!!
I think it is really the change of the ankle..
Cronin Knoll
I need more schooling.
Patsy Crest
I love plasmooons
Plz send spr multi analyte detection sensing pdf my email ishveer123143@gmail.com
if only she could narrate like an actual normal person and not like a corporate advertisment robot , it would be awesome
plazmonik biyosensörler tez konum için buralara geldim help me arkadaşlar
Hallo
My name is yuda
her voice makes me believe that she is a cute person