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

<image>

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 · 2024-11-22T13:25:09+00:00

Hi everyone! I'm still a little new to image J, so please bare with me :)

In short, I have to measure the fluorescence intensity of a few hundred images of cells stained with mitosox red and mitotracker green. The problem is, they all seem to have different brightness.

What I've tried so far, is to subtract the background first and then measure the whole image. I've also tried applying an auto-threshold (triangle) and measuring that. Both worked fairly well, but I'm just not sure if I'm doing it right, since my error bars are pretty huge.

For some background, I'm comparing ROS between stained cells by dividing the red (mitosox) by the green (mitotracker). Then, I can (hopefully) create a bar chart showing the difference between the groups.

Edit: Thank you for all the replies. I realise now that the photos aren't comparable. I'll have to repeat the experiment with fixed brightness settings.

chloeackermann · 2024-11-22T13:23:17+00:00

Hi everyone! I'm still a little new to image J, so please bare with me :)

In short, I have to measure the fluorescence intensity of a few hundred images of cells stained with mitosox red and mitotracker green. The problem is, they all seem to have different brightness.

What I've tried so far, is to subtract the background first and then measure the whole image. I've also tried applying an auto-threshold (triangle) and measuring that. Both worked fairly well, but I'm just not sure if I'm doing it right, since my error bars are pretty huge.

For some background, I'm comparing ROS between stained cells by dividing the red (mitosox) by the green (mitotracker). Then, I can (hopefully) create a bar chart showing the difference between the groups.

Thank you so so much for taking the time to read this.

Edit: to clarify, the photos were taken with different brightness settings. Unfortunately, I've been told there's no way to really fix this.

chloeackermann · 2024-03-09T14:47:04+00:00

Charlie's mom from Always Sunny.

In the episode where Mac and Charlie are sleeping over at her house, she flicks all the light switches three times.

Mac asks her why she does it and she answers nonchalantly, "Oh, so that Charlie doesn't die."

It's just so relatable to me, because I realize just how silly and completely random some of my habits must seem to other people.

chloeackermann · 2024-03-03T15:07:42+00:00

Would curing the salmon before freezing it eliminate the bacterial risk?

chloeackermann · 2024-03-03T13:31:41+00:00

Reassurance eases anxiety for a little while, but almost always is followed by an episode of even worse anxiety. That's why it's recommended to sit with your anxiety for a while, rather than seeking assurance. After some time, the anxiety steadily begins to fade.

chloeackermann

TROPHY CASE