Obtain cells and Resazurin Assay

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experimenter
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Obtain cells and Resazurin Assay

Post by experimenter »

Does anyone help me find resources on where I can order lung epithelial cells? How much would it be? or can I simulate this?
As my project is conducted in vitro, I want to be able to determine if the compounds I am using can be safe for humans. Therefore, I want to replicate the human tissue and assess the cytotoxicity.

Also, I am having a hard time understanding the Resazurin Assay. Can someone help?

Thank you!


Moderator Note: I've merged your two posts so the experts can respond to them together. Thanks.
pharrast
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Re: Obtain cells and Resazurin Assay

Post by pharrast »

The resazurin is simply a dye that makes it possible to see the level of activity of important enzymes, which give you an idea about how much chemical reaction by living organisms is taking place. So it basically tells you how "alive" is the stuff in your dish. Using this assay I think requires either a colorimeter or fluorescence detector, which you would probably have to use in a real laboratory.
As for purchasing cells, you may be able to buy human cells for 100s of dollars, but again it would be a waste if you do not have the necessary equipment to keep them alive. Unless you are working with a lab or university that has growth media, incubators, low temperature freezers, etc. all your cells will die without any results. I'm sure the lab you are working with can help you order cells. If you are planning on doing this experiment at home, I would suggest testing your "compounds" on plated bacteria and using simple colony counting as your readout.
experimenter
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Re: Obtain cells and Resazurin Assay

Post by experimenter »

Thank you!

Are there any labs that will work with students for a week or so to conduct the experiment, at least in the fall?
Any clues on how I can try to access a lab?
MS15
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Re: Obtain cells and Resazurin Assay

Post by MS15 »

The best option is to write to scientists in local university labs near you. Most labs have webpages that you can find on the university website, and these include emails of the lead scientist. You can write to labs which work with human cell lines, explaining your project and asking for help. I agree with the previous expert that if you want to actually do the experiment with human cell lines, finding a host lab is the best option. Please keep in mind however that many research labs are now working at very low capacities so they may not have the ability to help even if they want to.
Otherwise, the suggestion of testing the compounds on bacterial cells is great too - from bacteria to humans, most basic cellular pathways are the same so many compounds will inhibit the growth of bacteria as well, unless it is specific to eukaryotes in its mechanism of action.
experimenter
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Re: Obtain cells and Resazurin Assay

Post by experimenter »

Thank you!
experimenter
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Re: Obtain cells and Resazurin Assay

Post by experimenter »

Any ideas on how/ where I can purchase a Fluorescence reader for a price under $200.
experimenter
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Feedback

Post by experimenter »

Hello,
I would like to know if I can find an experienced biology/ medical sciences/microbiology expert to critique my project before I submit them to ISEF?
MadelineB
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Re: Obtain cells and Resazurin Assay

Post by MadelineB »

Hello Experimenter,

I've merged your most recent request with the thread from your posts regarding obtaining cells and the resazurin assay.

Even if your current project has changed, the experts who were helping you then will be willing to review your project.

Best of luck at ISEF!

Madeline
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MadelineB
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Re: Obtain cells and Resazurin Assay

Post by MadelineB »

Hello Experimenter,

I am not a microbiologist, but I've judged at ISEF since 2011 and I will be glad to review your slides - not for the microbiology content, but for the overall outline and flow.

Maybe you could attach a pdf of the 12 slides which ISEF 2021 is considering for your "presentation materials"?

Thanks,
Madeline
experimenter
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Re: Obtain cells and Resazurin Assay

Post by experimenter »

That will be really helpful!
How can I send my slides to you?
MadelineB
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Re: Obtain cells and Resazurin Assay

Post by MadelineB »

You can attach an acrobat PDF file to your reply. Look for the button labeled "attachment" in the bottom left corner just below the window where you type your message.

I'll keep an eye out for your message and the attachment!

Madeline
experimenter
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Re: Obtain cells and Resazurin Assay

Post by experimenter »

Is it possible to just send it to you only directly?

Thank you so much!
MadelineB
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Re: Obtain cells and Resazurin Assay

Post by MadelineB »

Hello Experimenter,

Science Buddies' guidelines are that all communication is through the Ask the Expert forums.

Are you having trouble using the "attachments" button?

You can also attach your file by dragging and dropping it directly in the message box when you reply to this message.

I'll be looking for your pdf file!

Madeline
experimenter
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Re: Obtain cells and Resazurin Assay

Post by experimenter »

Thank you so much and sorry about the delay in response. For some reason I did not get a notification about your reply.
I do see the attachment tab here.
Once you download it on your end, are you able to remove the attachment?

Thanks again.
MadelineB
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Re: Obtain cells and Resazurin Assay

Post by MadelineB »

Hello Experimenter,

I will check.

Madeline
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