Science Buddies: "Ask an Expert"

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.

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.

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?

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.

Thank you!

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

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?

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
Moderator

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

That will be really helpful!
How can I send my slides to you?

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

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

Thank you so much!

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

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.

Hello Experimenter,

I will check.

Madeline

Thank you! I will wait for your confirmation.

or, can I send my slides to [email protected]

ScienceBuddies is set up for you to communicate with the experts via this forum.

I suggest you sent your slides as a pdf file as an attachment, so I can start reviewing and helping you. I think you are going to run up against the ISEF deadline very soon.

Looking forward to seeing your slides.

Alright. Here it is. Please let me know.

Thank you so much for your time.

I've downloaded it. Perhaps you can delete the attachment yourself.

Warning: I have meetings the rest of today and evening so I will not be able to review this until tomorrow.

In the meantime, you might want to check the ISEF instructions for the presentation. I think they are pretty explicit about whether to include your procedures?

Thanks for uploading this. I'll respond by tomorrow evening.
Madeline

Thank you so much!

I will double check on the ISEF website as well for the instructions.

Looking forward to your comments and suggestions.

I skimmed through the first couple slides during my lunch break ... you need to follow the ISEF instructions for your slides - I believe the minimum font size is 14 pt. Check the other instructions ... I don't think they are optional.

More later ...

Hello Experimenter,

Remember as you read my comments, that I am not a microbiologist. I am a biostatistician/data scientist with experience working with medical researchers ....

This looks like a careful study with important results. However, even non-statistician judges will probably expect to see some of the statistical detail I note below!

1. Comments on your statistical design and analysis.

One major concern is that you do not describe the number of trials/observations at each of the treatment groups.

Similarly, I don't see that you describe the number of observations in your evaluation of the association between CFU/ml and Relative Fluorescence.

With respect to the motility experiments (Table 4), it would be helpful if you specified the treatment given.

The one statistical analysis which you report is a one-way analysis of variance. That is evaluating whether there were differences among the ?treatment groups. In the text below that table, you state different null and alternative hypotheses. I don't see how that test relates to your hypotheses as stated in the box just above the Methods section. I suggest that you consider using statistical analyses which provide appropriate tests of those hypothesis.

2. Other Comments:

I suggest that you explicitly label the box with what appear to be your hypothesis.

In your text, you use the phrase "proving the first hypothesis to be correct". Phrases like "rejected the null hypothesis" or "accepted the alternative hypothesis" are a better reflection of the statistical analysis. Science buddies has a project guide which might help explain how to phrase your hypotheses (null, alternative) and communicating your resultsl

https://www.sciencebuddies.org/science- ... ience-fair

3. And, as I mentioned in my 2 previous posts, you need to carefully follow the ISEF guidelines for the format of your slides, including the minimum font. This may sound picky, but I think you should expect that they will be picky!

Let me know if you have any questions about these comments and Good luck at ISEF!

Madeline

Thank you so much! I reviewed your comments and will make the necessary changes.

I also had a couple of questions regarding your comments...

1. Can I simply put somewhere on the board that "triplicates were done for all experimental trials"?

2. Was there a problem with the one-way ANOVA test? I tested if there was statistical significance between the independent groups (the individual and combination treatments) compared to the control group. I also explained my null and alternative hypothesis in the discussion slide; I don't think I understood this comment.

Thank you very much for your time!

Hello Madeline,

Your feedback on my ISEF slides were very helpful last year.

Do you have a moment to look at my slides from this year and provide any feedback?

Thank you!

Hi Experimenter,

I'm glad my comments were helpful! I'll be glad to look at your slides for this year. I won't have time until about this time tomorrow, so if you are hoping that you (or I) delete the attachment as soon as I've viewed it, you might wait until then!

I'm looking forward to seeing your slides!

Madeline

Thank you so much for your response. I will upload my slides tomorrow evening!

Thank you so much for your response. I will upload my slides tomorrow evening!

Hi Madeline,

Can I upload my slides now?

Hi Madeline,
Please let me know when you get a chance. I will upload my slides.

Thanks again!