Observe pH Damaged Cells Via Microscope?

[chemisthypnos]

For an upcoming research project, I intend to apply different solutions to cultures of cells. How would I determine if the cells are incurring damage from the solutions due to the pH of the solutions as I observe them under my compound microscope?

 

[BillTre]

Depends on what you want to look at.
If you are looking for cell death, you could use a fluorescent live-dead stain, such as this one.
There are many other ways to visualize live v.s dead cells.

There are also lots of other labeling reagents for visualizing other cellular conditions.
Here is a company that makes thousands of fluorescent reagents.

 

[chemisthypnos]

Depends on what you want to look at.
If you are looking for cell death, ou could use a fluorescent live-dead stain, such as this one.
There are many other ways to visualize live v.s dead cells.

There are also lots of other labeling reagents for visualizing other cellular conditions.
Here is a company that makes thousands of fluorescent reagents.

Thank you very much for your expedient response. The purpose of the research is to evaluate the effects of a solution on cells for a physiological application. I would like to pose 2 scenarios as this is my first time doing any kind of physiological research.

1. After testing with the solution, cells are retrieved and are found to be mostly dead. Is this safe for use with the organism? (I don't think so!)
2. After testing with the solution, cells are retrieved and are found to be alive but seem to be damaged. Is this safe for use with the organism?

 

[BillTre]

It is difficult to give any more appropriate response without knowing more details of:

• What kind of physiological effects you are interested in?
  
• What the organism you talking about is?
• When you say safe for the organism, what does this mean?

 

[chemisthypnos]

It is difficult to give any more appropriate response without knowing more details of:

• What kind of physiological effects you are interested in?
  
• What the organism you talking about is?
• When you say safe for the organism, what does this mean?

1. I am looking to see if the solution being tested causes notable damage to an organ (the solution is advertised as being physiologically safe but I am investigating if this is actually true).
2. The organism is a mouse
3. I would define it as not causing pain or damage to the organ such that it impairs the organ's structure.

 

[Ygggdrasil]

Here's a good resource on assays to assess cell viability and cytotoxicity of various compounds: http://www.abcam.com/kits/a-guide-to-cell-viability-proliferation-and-apoptosis-assays

Testing cells in culture would not be sufficient to say whether a compounds would cause toxicity in mice. For example, if the compound itself does not cause toxicity but is metabolized in the body to a compound that is toxic, testing the compound in cultured cells would not likely reveal this toxicity. However, testing toxicity in cultured cells is a good first step to perform before testing anything in mice.

 

[chemisthypnos]

Here's a good resource on assays to assess cell viability and cytotoxicity of various compounds: http://www.abcam.com/kits/a-guide-to-cell-viability-proliferation-and-apoptosis-assays

Testing cells in culture would not be sufficient to say whether a compounds would cause toxicity in mice. For example, if the compound itself does not cause toxicity but is metabolized in the body to a compound that is toxic, testing the compound in cultured cells would not likely reveal this toxicity. However, testing toxicity in cultured cells is a good first step to perform before testing anything in mice.

You raise a very good point, Ygggdrasil. This is the solution being tested: https://envirofluid.com/worksafe-environmental-chemistries/scale-calcium-removal/triple7-enviroscale The plan is to expose the organ of interest of the mouse to the solution and take tissue samples every few minutes. I just want to see if the solution causes pH or other types of obvious damage to the cellular structure or if it kills cells at the exposed regions of the mouse. Do you think that this kit would be useful for that?

 

[Tom.G]

The SDS (Safety Data Sheet, https://envirofluid.com/worksafe-en...-calcium-removal/triple7-enviroscale?doc=1242) for that product indicates it as "Hazardous" on pg 7, and pg 5 states it has a pH of 2.1, which make it stronger than Sulfuric Acid at pH 2.75. Be careful.

 

[chemisthypnos]

I am quite glad that you brought this up, Tom.G. I found it bizarre to an extreme that it states on the SDS that the product is non corrosive but has a pH of about 2.1. Also, on page 6, it states that the most that it will cause is some redness or skin inflamation. That is not what I would expect to find on a substance that has a pH of 2.1. Compare the SDS for the aforementioned substance to this one for sulfuric acid: https://www.msdsonline.com/2014/07/22/sulfuric-acid-safety-tips-sulfuric-acid-msds-information/ The envirofluid SDS doesn't even make a big deal about it getting in the eyes as it would only cause blurred vision, redness, etc... Unlike sulfuric acid which will cause blindness.

I find this most perplexing so I wanted to investigate. Any ideas for why this may be?

 

[Tom.G]

Sulfuric Acid is also highly reactive with water, which is part of its danger. Since that Envirofluid is water based and diluted to 4-5%, it is probably not as reactive. To add a bit more perspective, Lemon Juice has a pH around 2.0 and White Vinegar(5%) around 2.4. Since the constituents of Envirofluid are unknown, we really have no way of telling how reactive it really is without testing. I would guess it to be somewhere around Lemon Juice... but until that is known I would treat it as Sulfuric Acid.

To answer your direct question as to why, Marketing Hype?

Please let us know what experimental results you get.

Cheers,
Tom

 

[chemisthypnos]

To pose one final question, I am thinking of going with a cell viability stain (to see if they are alive or dead). These stains are visible under a compound microscope, right? Thanks.

 

[BillTre]

To pose one final question, I am thinking of going with a cell viability stain (to see if they are alive or dead). These stains are visible under a compound microscope, right? Thanks.

Yes.
There are, as I understand it, several different kinds sold by different companies.
There is also non-fluorescent versions, like the old stain trypan blue, that stains dead cells.

 

[chemisthypnos]

Thank you, everyone. I think that the information that you have provided will help me greatly. Have a great Summer.

 

[TeethWhitener]

stronger than Sulfuric Acid at pH 2.75.

This is not how pH works.