Making Fluorescent Yeast Exciting

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TheWolf2978
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Joined: Wed Jan 17, 2018 4:20 pm
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Making Fluorescent Yeast Exciting

Post by TheWolf2978 »

Hi! I’m a grade 7 student in an advanced program, and I’m participating in my first science fair. For my science fair project, I will be editing the DNA of yeast to make it fluorescent using CRISPR cas9 and running tests on different samples and I hope to measure the difference in the fluorescence of the yeast. I’m using the Odin yeast CRISPR kit, the instructions are here:http://www.the-odin.com/engineer-any-ye ... uoresce-1/.
(So far) I have 3 questions. First, are there any ways to measure the fluorescence of the yeast (either with actual numbers or variables), and if so how could I do this at a reasonable price in a relatively short period of time? Second, are there any ways to photograph fluorescent yeast?
Third, even though I think that I have a pretty cool topic, I don’t know how to create the wow factor that will really impress judges. Thank you!
SciB
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Joined: Fri Feb 01, 2013 7:00 am
Occupation: Retired molecular biologist, university researcher and teacher
Project Question: I wish to join Scibuddies to be able to help students achieve the best science project possible and to understand the science behind it.
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Re: Making Fluorescent Yeast Exciting

Post by SciB »

Hi,

First off, please tell me what sequence you will be inserting into yeast to make the light. The mode of light production will decide what method you would use to record it.

Fluorescence, bioluminescence, chemiluminescence and phosphorescence are all modes of light emission but they each operate differently. Fluorescence, for example, like the light emitted by the commonly used green fluorescent protein (GFP) only lasts for nanoseconds after the exciting light is turned off. Phosphorescent materials such as zinc sulfide are used for glow-in-the-dark toys and other items. These substances absorb the energy of the activation light just as fluorescent materials but emit light over several minutes.

There are ways to measure fluorescence in the lab, but you would need a specialized instrument called a spectrofluorometer to do that. If the sequence you are inserting is for GFP or one of the other fluorescent proteins then you will have to use a fluorometer to measure it.

Bioluminescent organisms like dinoflagellates and fireflies use a chemical reaction--NOT fluorescence--to make light. Light is produced when luciferol is cleaved by the enzyme lucferase. The glow of light that you see when you break a light stick, is produced by a different chemical reaction. Since the light is not biologically produced it is called chemiluminescence.

There are ways to photograph the relatively dim light of luminescence with a smart phone and then measure its intensity using an image analysis program. You can readily find information about these methods online.

Let me know more about your procedure and maybe I can make some suggestions that you can try.

Sybee
SciB
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Posts: 2066
Joined: Fri Feb 01, 2013 7:00 am
Occupation: Retired molecular biologist, university researcher and teacher
Project Question: I wish to join Scibuddies to be able to help students achieve the best science project possible and to understand the science behind it.
Project Due Date: n/a
Project Status: Not applicable

Re: Making Fluorescent Yeast Exciting

Post by SciB »

I forgot to check the link you provided.

So, now I see the inserted sequence is for green fluorescent protein, GFP. Reading the initial information, I also see that the GFP can be made to fluoresce by continuous illumination with an ultraviolet light source. You can set up the UV light source on a stand so that the Petri dish on which the yeast is growing can be continuously illuminated. A camera on a tripod can be used to photograph the emitted light. You will have to do some checking online to find out if your camera or smart phone is capable of photographing fluorescent light.

In order to measure the intensity of the light you can use a free app called ImageJ: https://imagej.nih.gov/ij/index.html
If you have questions about how to use it let me know and I will guide you through it.

Sybee
TheWolf2978
Posts: 2
Joined: Wed Jan 17, 2018 4:20 pm
Occupation: Student

Re: Making Fluorescent Yeast Exciting

Post by TheWolf2978 »

Hi Sybee, I just wanted to say thank you for the information, it was very helpful. I will be sure to check out the app. This weekend, I have been working on the procedure and it has been going well. I will be sure to ask you if I have any more questions. Thanks again!
SciB
Expert
Posts: 2066
Joined: Fri Feb 01, 2013 7:00 am
Occupation: Retired molecular biologist, university researcher and teacher
Project Question: I wish to join Scibuddies to be able to help students achieve the best science project possible and to understand the science behind it.
Project Due Date: n/a
Project Status: Not applicable

Re: Making Fluorescent Yeast Exciting

Post by SciB »

You are certainly welcome! Glad it helped. If you have any questions about understanding the science, be sure to post again. We want you to be able to explain clearly what is happening in your project and CRISPR technology can be a little challenging to put into simple language.

Good luck!

Sybee
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