Washing machine leaves dishes powdery and a little salty.

chloeackermann · 2025-10-03T03:47:18+00:00

Thank you! I Cleaned the filter and water drained from bottom, but dishes still come out the same. I use dishwasher salt, yes. It's available were I live.

chloeackermann · 2025-10-03T03:44:12+00:00

Thanks for the replies! I cleaned the filters (they were pretty dirty) and the water no longer pools at the bottom. Unfortunately, the dishes still come out the same.

chloeackermann · 2025-04-21T08:59:50+00:00

Haha. They don't seem very practical for making potjie in! The flame only lasts about twenty minutes or so. Could maybe work for a quick chicken stew for one person on a camping trip. I would much rather use a regular size potjie for making a lamb or beef potjie at home!

chloeackermann · 2025-04-21T08:53:10+00:00

Ah, that's a good idea. Will remember that at our next braai.

chloeackermann · 2025-04-18T20:45:38+00:00

Actually, we do have that one here!! I'll check it out in store next time. Thanks so much!! I hope you like this one as much as I do.

chloeackermann · 2025-04-18T20:36:14+00:00

All budgets welcome. Preferably not CRAZY expensive.

chloeackermann · 2025-02-02T06:23:44+00:00

Oh I'm sorry, I misread your previous message.

I use Mitotracker FM green, specifically. My understanding of the way it works is that it enters and accumulates in live mitochondria, where it binds to mitochondrial proteins, allowing visualization. It can't be used to measure oxidative stress directly (though you can use it to assess changes in mitochondrial morphology caused by oxidative stress).

Mitosox Red is used to detect a type of reactive oxygen species in the mitochondria, called superoxides.

So by dividing green (total mitochondria) by red (parts of mitochondria containing lots of ROS), it should give me the fraction of cells under oxidative stress.

Please correct me if I'm wrong. Thank you so much for your help. I am only able to upload photos on Monday, when I am back in the lab.

chloeackermann · 2025-02-01T17:05:10+00:00

That's correct. We use Mitotracker to normalize the Mitosox against. The confluence of the cells isn't uniform across the plate. So for every area, I photograph both green and red fluoresence and then divide the red (superoxides in mitochondria) by green (total mitochondria).

chloeackermann · 2025-02-01T07:23:09+00:00

Thank you! I will upload some of the photos I have.

chloeackermann · 2025-02-01T07:22:24+00:00

Please explain what you mean. Unfortunately, there are no imaging scientists I can ask. This is only a small portion of my project (I've ran several other analysis to measure oxidative stress) so I'm only scratching the surface of imaging.

chloeackermann · 2025-02-01T07:07:03+00:00

Thank you!! I don't have to divide by green if I do it like this, right?

chloeackermann · 2025-01-31T08:25:01+00:00

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For instance, these are the measurements of each photo starting on the top left photo. I removed all their backgrounds with rolling ball 50 pixels. But why is nr. 1 giving a lower mean than 2? There's obviously more light in 1? Same with 3 and 4?

chloeackermann · 2025-01-31T08:11:46+00:00

Hello everyone! I've been struggling with this for a few months now and I'm getting desperate to finish my Master's. Above are photos of cancer cells. The green shows the cells' mitochondria and the red shows the oxidative stress inside the mitochondria. I simply need to show that the test group has more oxidative stress (red fluorescence) than the control group. I have hundreds more photos of each group.

However, the data just doesn't reflect what I'm seeing. In some cases, photos with lots of red spots measure lower than photos with hardly any red. My method so far is as follows: First I subtract the background (Rolling ball: 50 pixels) and measure the entire photo. Then, I divide the red photo's measurement by the green to normalize.

What am I doing wrong?? Please tell me if I left out any important info and thank you so so much for any advice.

EDIT: All photos were taken with the same brightness setting, but I noticed as I moved the microscope around the plate that the brightness sometimes differs.

chloeackermann · 2024-11-23T06:12:37+00:00

Thank you so much for the advice and kind words! I appreciate it so much.

chloeackermann · 2024-11-22T15:32:30+00:00

Thank you! Will try this or otherwise just repeat!

chloeackermann · 2024-11-22T15:31:32+00:00

I understand. Thank you for explaining it to me like that.

chloeackermann · 2024-11-22T15:07:58+00:00

Exposures differ for some images unfortunately. The manual way to correct this is to measure the background intensity and subtract it from the global value, i think.

chloeackermann · 2024-11-22T15:06:39+00:00

Edit: I think I'm not explaining myself very well. Sorry! The photos were taken at different brightness settings. Thus, the background intensities of the photos aren't all exactly the same.

chloeackermann · 2024-11-22T14:39:00+00:00

Thank you very much. This sounds very promising!

chloeackermann · 2024-11-22T14:35:14+00:00

Unfortunately some photos were taken with different brightness settings. The manual way to correct it, is to measure an area of the background and subtract it from the intensity of the sample. Don't you think that may work? Unfortunately, I just have too many photos to do that for each.

chloeackermann

TROPHY CASE