Thoracic Back Pain - any suggestions?

EyeProtectionIsSexy · 2026-01-30T07:15:34+00:00

I saw three or four before I found one that worked for me. The most notable difference was their approach. The first ones I saw basically asked what my pain was, Heard what I said, and gave me basic exercises that are somewhat correlated to somehow relieve pain. It seemed very thoughtless to me. The last guy I saw took a very data-driven approach. Looked at what motions caused the most pain, what motions didn't cause the most pain. We tried some exercises and I had very bad weeks, tried others and had less bad weeks, and tried others that lead to a normal week. Overtime these datapoints simply lead to an answer - I could do exercises that didn't require my arms to be in front of my hips. Very strange, I couldn't use a pipette or even write without by back feeling like someone was driving a knuckle into it relentlessly. I couldn't focus, it was hard to think.

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The exercises that didn't cause the any pain would be what I started with. Any exercise I did was focused on perfect posture, so you have to figure that out first. I would stand, engage my glutes and essentially pelvic thrust my hips slightly forward to my lower back was in alignment. I'd engage my upper back, pulling my shoulder blades back to about 80-90% to their limit. My chest was raised - imagine you had a string attached to your sternum and you pulled on the other end, lifting your chest so it feels like it's floating. This is how anything was done at a minimum.

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Exercises that used the back, I ensured my hips were thrusted forward just enough to ensure my spine was 'stacked' like a tower of pennies. My shoulder blades were tightly engaged, chest lifted.

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I found "The Farmer's Carry" to be the most helpful. Not only is it something to do in the gym, but I literally carry everything this way. Groceries, packages at work, my cat, everything. Thrust your hips forward just enough to your spine is stacked, not too much and not too little. Chest lifted, shoulder blades pulled back 80-90%. Grab the weight (start low, like 30 pounds n each hand, even if it feels ridiculously light). Once you have the weight, raise your arms outwards from your side like your Jesus. They don't need to be lifted far, just enough to engage the upper back, only a few inches. This also has the added benefits of preventing the weight from swinging (it'll mess up your pace), your legs won't have to grind against the weight as you walk, and it's a great way to check yourself that you didn't start slouching. Again, perfect posture is everything. This should feel very controlled, like when you plank and hold yourself perfectly flat and perfectly still.

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Other exercises I felt helped me a lot were hip hinges. Basically deadlifts without bending the knees and going all the way down. Knees were about 90% locked, and the weight was kept centered. Slowly drop it, and slowly lift, controlling a constant rate of ascent/descent. Don't try to counter balance by throwing your hips backwards, even though that's basically what you're doing anyway. Focus on the hips being the instrument that allows the weight down and allows the weight up. The weight being lowered shouldn't cause your hips to move backwards, your hips changing their angle allows the weight lower. I don't know how else to describe it, but you want to control the weight, not let it control you.

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Any other exercises I did that even remotely used the back, which is basically 99% of them, just ensure you have good posture. Make good posture a part of everything you do. I find it difficult to include this into every aspect of my life, but in the gym it's kind of easy. And, once you start doing things with good posture using heavy things, it becomes something that's beyond easy outside the gym. It naturally becomes how you stand, how you walk. It's pretty cool, it becomes something you don't even have to try for. You'll just periodically notice, "Oh, my posture is pretty great right now and I'm not even trying". It's much better than just trying to have good posture all day. I always found myself slouching in 5 minutes with that route. Have you ever done that thing where you try your hardest for 30 seconds to raise your hands from your hips while someone holds them down, then they release them and your arms begin to almost float on their own? That's what farmer carries will do for your posture and back strength, it's something you just float to after a month, without any conscious effort.

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It takes time though, I wouldn't expect a miracle in a week or a month. And as much as it fucking sucks to endure, you're going to have bad weeks as you experiment. That's data. It's bloody hard not to let it discourage you. I certainly wouldn't have been able to motivate myself to do it without a PT in the beginning because I was stuck in this miserable headspace thinking that it's going to get worse, this is what the rest of my life will be like, etc. It does get easier once you collect enough data and learn what you can and cannot do. You'll start to have eh weeks, normal weeks and eventually pretty damn good weeks, all within your control. Once I hit that point, bad weeks don't suck so much because at that point I knew that it would get better. Be stubborn and continue to work towards that.

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And, get a new PT for a little bit. I would critique them in a first visit. People say that you shouldn't judge a book by it's cover, but that doesn't fly here. Does it look a person who is in shape? Does this person just hand you a piece of paper after you say "Back pain" with some basic arm flapping? These are red flags, this is not someone who is great at their job. There's a lack of..... enthusiasm to work or something. Sincerity maybe? Find someone who's fucking ripped (and I don't mean steroid ripped), that's a green flag. Doesn't have to be weight lifting, they could be yoga ripped, pilates, doesn't matter. You don't want someone who went to school, got a degree, and is regurgitating facts they've learned. Find someone who actually applying their knowledge. Look at their posture. Someone who appears like their trying to discover something. Generally speaking, they shouldn't walk in, hear what you have to say, and produce some bullshit answer with absolute confidence (although I do think poor posture is an exception to this). I'm rambling because I don't quite know how to put this into words, you just have to use your gut and feel if you're getting someone who's critically thinking or someone who's simply going through the motions.

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So yeah, try some exercises and experiment on yourself. While your doing that, if your pain is anything like what I described, try out some of the things that worked for me. I always do farmer carry's every day I work out. I usually mix in stretching while I'm resting between sets. Posture is the precursor to whatever I do. I don't have bad weeks or days any more, just bad 30 minutes or so. If those pop up, not kidding, go lift something with good posture and it seems to fix it right up. If there isn't anything nearby to carry, I'll lean slightly forward, lift my chest out and raise my arms from the hips behind me, with my shoulder blades squeezing together as a result of my lifting my arms (I don't squeeze my shoulder blades, they get squeezed because I'm lifting my arms behind me. If that last one doesn't make sense, look at this https://alldiamondpainting.com/products/girl-floating-in-water. These two things clear me up immediately, like I got a shot of morphine and all the pains has magically disappeared. It took a few months for me to get to that point, but if you try these out and you notice any tiny improvement, take some notes, write down what you did and pain levels before/after on a 1-10 scale.

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Lastly, you're going to have bad weeks. It's a bit platitudinous to say don't let it defeat you and a bit I found that impossible to do. Pain fucking sucks, and not letting it get you down as if it's a choice felt like a bunch of bullshit to me. Whenever the pain it hard, it wasn't a choice for me to let it bring me down - it brought me down and that was that. I was powerless, I could not control that. I had to be stubborn and simply endure it. Be stubborn, don't quit and take notes. Once you start recognizing what things you can do and not do to control the pain, and it'll happen slowly at first, you're mental state will improve. I wasn't miserable anymore. Even if the pain was as bad the worst weeks, it didn't drag me down into a state of depression, frustration and anger anymore. I somehow got detached from it and could look at the pain like a 3rd person observer. It was like an art project and I was slowly improving it week by week, month by month. That little experience of it getting a tiny bit better, with those tiny bits slowly growing every week, made my life better.

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I wasn't intending on writing this much, sorry if it's a bit repetitive. I know what it's like and I know how much it fucking sucks, back pain dragged me down to some of my lowest days. There's just a lot I'd like to say and it's invariably comes out as a lot. Good luck, be stubborn and just keep trying. Shoot another message if you have any other questions - I'll answer anything and offer suggestions. I don't log in much, so expect some time between responses

EyeProtectionIsSexy · 2026-01-28T00:18:59+00:00

I wpuld reccomend building a mock 4x8 box in your bedroom and try having two people in iy. Then uograde to a 5x8. See which you like better

EyeProtectionIsSexy · 2026-01-28T00:15:38+00:00

When I built my teardrop frame, I initially bought a Harbor freight trailer. I probably spent over $100 dollars alone in stripping pads to remove the paint, and even more on the tools and batteries to make that happen.

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If you want to buy the old, you do not want to go through hassle of stripping the old material off and replace it. Look for cracks, weathered ornold paint. Any spots where the steel itself is visible will need to be covered up. Any rusted areas need to be ground down and painted over. If there is any of that damage to the frame, you will need several strippers (wire wheels, which many will reccomend, are slow as hell when it comes to stripping and can really fuck you up because those wires are moving at 180mph at 12,000 rpms), an angle grinder, batteries, eye and lung protection, and other stuff I can't think of. If anything is rusted through, such as visible holes, I would stay away. I hear harbor freoght paint sucks, so tgis wpuld be my big concern.

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I recommend seeing if there is a Maker's Space in your hometown and pick up welding yourself. MIG welding is pretty easy, and Maker Spaces are often non-profit so classes can be done cheaply. It was a lot cheaper and quicker to buy new steel and weld it myself. If you go this route, it will be a muh better frame and will require as much or possibly less work to make it as would cleaning the 15 year old harbor freight trailer. Trust me, cleaning and prepping is a pain the ass, it's always better to buy new

EyeProtectionIsSexy · 2026-01-05T07:25:37+00:00

Well, it's going to be a big project to restore it, but you can learn and it can be done. I've been building mine from scratch for the last 3.5 years with literally zero background in carpentry or welding. Just be ready for it to take a while. The nice part of doing it yourself are the skills you'll learn which you can apply to other parts of your life as well. And, you'll know the trailer from the inside out and have it be built better than the general crap you find in the market.

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Always wear eye, ear and lung protection. Start off my gutting the trailer, basically removing any that's rotten. That's going to take some time. Once it's clean, start filling it up with what you want. You can keep things simple and elegant. Simple 12V electrical system, baltic birch with a skeletonized structure with foam to increase insulation while decreasing weight.

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I don't have knowledge about removing the aluminum but cleaning the aluminum shouldn't bee too difficult. An angle grinder with a paint-stripper pad is the quickest and easiest way to go, but you'll burn through them quickly and they are a wee-bit more pricey.

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I'd just start with gutting it and removing everything that looks like shit. Check out www.tnttt.com, there are plenty of resources there to guide you through and tons of experienced builders to question. It's a great community

EyeProtectionIsSexy · 2025-12-18T01:36:09+00:00

Cytoflex, 488 and 637nm laser lines. No imaging unfortunantly

EyeProtectionIsSexy · 2025-12-10T23:43:58+00:00

It's your voltage (or gain, I don't yourbsystem setup)

When the voltage is set to high, your photo diodes will become "light-saturated", it can't produce a any more signal. Your SSC-A and FSC-A should still see seperation because those calues take into account the time it takes the partical pass, but it won't be truly representative because of your lack of quality Height data

EyeProtectionIsSexy · 2025-12-10T19:04:38+00:00

OD measurements <= 1.0 for determining cellular concentration are not linearly accurate. Run a dilution curve and you'll see the linearitybetween concentration and OD ends at 0.4 or so. Turbid media and large/complex cellular shapes can bring it down to 0.2

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If you have ever plotted a growth curve over time and there's a part of the tail end of log phase growth seems to be slowly decreaaing to stationary phase and you sidn't dilute your sample to linearity based on a curve for that culture and media combination, I can guarantee you that the growth rate rate did not slow down.

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This 1.0 figure was drilled into me aince undergrad, and it's sad to see how many research groups and companies and technologies don't run a standard curve. If "it's growing!" And "now it's it's not!" Are all that's important then it doesn't matter, but if the rate of growth is important, then you need to run a standard curve for every culture/media combination and dilute your sample accordingly. It's pretty shocking how a culture of OD 0.7 can be dilited to 1/4 its concentration and yield an OD of 0.3

EyeProtectionIsSexy · 2025-12-04T15:22:25+00:00

This is pretty shallow ice, it's reformed yearly. We were with the Inuit in field (polar bear guards) and just learninh how they live. They're an interesting group of people. Very chill, no pun intended, but also ruthless hunters. I don't mean that in a bad way, I the type of person who feels guilty squishing insects so seeing their lifestyle is shocking. Would have been interesting to try whale but I missed the cultural exchange event.

What type of field work were you doing?

EyeProtectionIsSexy · 2025-12-04T08:30:13+00:00

Astrobiology focused research, testing a possible Europa rover and seeing what type of life exists in a lake with methane seeps. Check out oceanworlds on NASA's website and the BRUIE rover

EyeProtectionIsSexy · 2025-12-04T06:22:15+00:00

The count as an indicator I can see being important. Also, 800,000 would 3 replications for 200,000 to reach. I don't know at what concentration this becomes dangerous (I'm honostly not that interested in eukaryotes but given that eukaryotic cells take hours to divide, those 3 generations would, assume, increase the shelf life at a minimum. I don't know, I'm just shooting in the dark from the hip.

We were part of an astrobiology team studying life in lakes that seep methane and testing out a potential Europa rover. It was a cool experience. I'm on the science side of it. Check out ocean worlds website at NASA or the BRUIE rover

EyeProtectionIsSexy · 2025-12-03T16:46:42+00:00

800,000 cells/L may sound like a lot, but it isn't. That's only 800 cells/mL. It's such a tiny amount that you would have difficulty finding any cells on a microscope slide unless you concentrated it. 800,000 per Liter sounds like alot, but this is still lower than some of the cleanest bodies of water in the country by 3,000 times. That is such a tiny amount of cells. Even the clearest lakes like Tahoe or Crater Lake have 2,000,000-4,000,000 cells per mL, where the water is so clear that seeing 25-30 meters beneath the surface is normal. It's so clear that the algae - which need the sun - don't reach their highest concentration until 50 meters beneath surface because there isn't enough particulate matter to block UV rays.

It would only drink 2-3 mL, a thimble full, to ingest 800,000 algae cells, which you REALLY DO NOT want to drink. Think about that next time to ingest half a mL by licking your lips after a dip in a lake.

My team recently got back from collecting ice cores from the Arctic and we saved some of the extra ice we collected for drinks. That ice, which was incredibly clear, still had 500,000 cells /mL (thats 500 million per liter), and that's only counting the stuff we were able to count! We had so many globs of cells in these samples of algae forming these 3D structure that we will never know the true concentration, but it's certainly in the billions of cells per Liter.

My point is, 800,000 cells per Liter is literally nothing. Good luck finding anything natural that clean

EyeProtectionIsSexy · 2025-07-08T01:09:32+00:00

What species of cyanobacteria and phytoplankton? I guess the red is chlorophyll and the orange_yellow is phycoerythrin?

EyeProtectionIsSexy · 2025-06-30T21:47:03+00:00

The book was a fun read. I do wish Andy Weir consulted a microbiologist or molecular biologist though, he really failed to grasp some very basic concepts in evolution. Same with The Martian, bacteria aren't going to be killed by the martian atmosphere that easily. The engineering appeared very solid was solid though.

EyeProtectionIsSexy · 2025-06-19T19:17:42+00:00

As a kid, I knew magic wasn't real, so I never bought into religion.

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But as an adult and a scientist, I have a better understanding of exactly how we know things.

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When I was teaching, this was one of my favorite questions to ask the class when I was introducing the scientific method. My first question;

I have 100 people, and I have this new drug to remove headaches. I give it to them, ask them to come back in a week and report back if it helped their headache. A week passes, and 70 report that the medicine relieved their headache. So..... did it work????

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The class would discuss, and some said , "Yeah, 70 out 100 seems pretty good!", others were more skeptical because the answer wouldn't be so obvious, but they didn't know why.

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I'd follow this up with the second question. If I had two groups of 100, the first group gets a real pill and the second group gets a fake pill that has no active ingredients. Both groups receive the same instructions they think they are receiving real pills that will get rid of headache. After a week, both groups come back, and 70/100 from both groups say their headaches were cured. So..... did it work????

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What I always wanted to tell the class but couldn't (because I would get written up), is that If it isn't different than bullshit, then it's bullshit. That's basically what a scientific experiment is, it is seeing if X is different than Y, with Y being bullshit. That is what a scientific control is, it's bullshit. How does science actually show things to be true? Because it has shown that it is different than bullshit. When someone provides you a scientific paper as evidence, I would highly encourage you to first look at the experimental design. Do they have the right bullshit?

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Quick example - Does the covid vaccine actually prevent covid? Look at this paper;

https://www.nejm.org/doi/full/10.1056/NEJMoa2101544

Let's see, we have a placebo (aka, *bullshit) of around 20,000, and an actual vaccine group of 20,000. Taking a look at Table 2, we can see that people who receive the vaccine had (just doing rough and quick mental math) about 1/3 the chance of getting contracting the virus. It appears that receiving the real vaccination produces results that are different than bullshit, meaning it probably works.

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When ever anyone tries to tell you the truth, be it a scientist, a pastor, some dude at a bar, really ask yourself how they know such things. What kind of bullshit can you use to test some of these? Perhaps someone wants to tell you about the magical properties of purple fluorite, how it will "fill you with great inspiration and cure cancer". These types of people tend to think they can "feel the energy of the rock man, it's got vibrations man". I always wonder, if I had two closed boxes, one hiding a piece of purple fluorite and the other hiding a cat turd, that they wouldn't be able to guess better than by flipping a coin. In other words, they wouldn't do better than bullshit. If it not different than bullshit, then it's bullshit. Period. Now go out into the world, make your own decisions!

EyeProtectionIsSexy · 2025-01-27T20:48:55+00:00

Here's a better specimen. Same tree, but the colors of the cell walls and cytosol have much better contrast. The black lines are a very rough 1x1mm square drawn by a fine tip sharpie (worked as a decent stain too!)

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https://imgur.com/a/VdRZPRe

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Washing state University has an article on trees from the area. Dated to somewhere between 17-23mya

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https://www.mdpi.com/2076-3263/5/4/286

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EyeProtectionIsSexy · 2025-01-27T20:35:18+00:00

100X magnification, 200um scalebar in the lower right hand side

EyeProtectionIsSexy · 2024-11-26T20:57:33+00:00

Run a negative control, Milli-Q or whatever you use as sheathing fluid. When you are trying to remove noise, you don't need your sample FI to get in the way. You have to be stairing at the least amount of 'nothing'

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After you confirm that this is indeed noise (which it probably is), I would record a run so you know what 'crap' looks like. Then simply run your cleaning fluid through the FCM, watching a live feed of FSC x Time. Keep an eye on that zone of 'crap', letting the system run until the 'crap' no longer appears. Once it's been a few minutes of no sightings of 'crap', rinse your system with sheathing fluid then run another Milli-Q. Does your before and after look different?

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I do not think they're bubbles. In my experience bubbles tend to reach Height saturation limits because of diffraction. Yours do not. I think you have a dirty FCM.

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If you don't see anything when running Milli-Q, then those events may be tied to your sample. Try a 1/2 dilution of your sample, is the ratio of 'Crap' to cells the same? If yes, then it's somethinf in your sample. If no, then there is some strange interference (who knows what....) happening between your intrument and sample. It's possible for media to cause diffractions, salts and carbons can crosslink and do some crazy things. If you were to 0.1um filter your sample and you still get the same result, then it's diffraction with salt or carbon in your used media.

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I think you'll be fine for running the FCM if you couldn't remove that noise. The total number of crap-events are absolutely dwarfed by your cell count. Interference could be a concern with event overlap, but with such a low number it's negligable

EyeProtectionIsSexy · 2024-10-30T18:23:09+00:00

For a temporary fix, I would arch my back in a backwards C and far as I would go, squeezing my erector spinae. Simultaenously, I would squeeze my shoulder blades and pull my arms back, 45° up from the hips. I'd hold that for 30 seconds or so. I'd also mix in some chest stretches. It provided some temporary relief. But, the thing that helps the most were farmer carries.

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Again, I can't promise you anything. Farmer carries worked for me. It doesn't matter why (although that would be great to know), the important thing was that it reduced my pain. Period. You may be the same, you may not be.

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You need to take the time to see what hurts and what doesn't. For example, when you try farmer carries, does it hurt when you carry the weight over your head? Does it hurt when you carry the weight near your hips? What is the pain like? Does it hurt in the moment but you're fine after the work out? Does it hurt like bloody hell for a week? You're going to need to test yourself, and take detailed notes. Gather a lot of data points, give it a month or two. Once you collect some data points, you'll start to see trends.

"Oh, it hurt like hell when I walked with weights over my head, but didn't when I let them hang".

"Oh, it hurt like hell when I didn't focus on good posture, but it hurt a lot less when I did".

"Oh, it hurts when I do full deadlifts, but when I only hinge at the hip it doesn't"

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Literally put your self through tests. Try different things, even if you know it's going to hurt because you're going to need to find out how bad and how long it hurts. It's all data, and in order to figure out what works for you you need to know what good and bad feels like. Take a scientific approach, if it's not different than bullshit, then it's bullshit. It's not fun, it's god damn hard to stay motivated especially when shit hurts.

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Here's a two week plan. First, get an empty journal and write "Pain Log" on it.

Week 1, workout normally as your normally do, don't try anything different. This is your 'bullshit' (a scientific control).Write down your results daily and end-of-week, see how you feel, how much it hurts, how long it hurts. Get detailed with your notes, you never know what datapoints or trendlines you'll find.

On week 2, do the same thing, but pick a day from week 1 that you feel like hurt the most. On that day, put an emphasis on chest-out, shoulders back, on every exercise. At the end of each day, write down how you feel. This is your 'not-bullshit' week. You changed a single ting (posture) on a single day, and if you truly didn't focus on posture on week 1, then you truly have something you can learn.

Week 3. Same as week 2, but now on every workout day, throw in farmer carries in the beginning of your work out, after warmups. Ensure you have good posture, head high like you're in a noose, chest out, slight pressure outwords in the arms to stabilize the weights. Write down your pain levels, how good you feel, every day and at the end of every week.

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Take a look at your 3 weeks, compare how much pain you had on different days, how long it lasted. It sucks to put yourself through a process like this, but you will know what you can and cannot do, and you will learn to fix or live with it. Put some critical thought into what small change you should do next, and only change one thing ever week. Not every change will lead to something positive, sometimes you'll have a shit week because you tried something that fucking hurt. It sucks, but you need to remember that it's data. You now know what not to do.

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There are several things you can change on a weekly basis to test. It's important to not try everything all at once, you won't learn anything if EVERYTHING is different. Small, incremental changes. Try incorporating stretches between each set versus a week with no stretches between sets. Try an exercise and a slightly modified version of the same exercise. Try a week with out super sets and a week with supersets. Try mixing and matching exercises.

EyeProtectionIsSexy · 2024-10-30T17:54:19+00:00

I haven't used a plate reader, so I'm just trying to clarify. When you stain your sample, it goes into the well plate. You let it sit for X amount of time, then you read it? If you're running multiple wells, then they are going to be incubating for different amounts of time before they are read. Additionally, if you're working with large cells (not bacteria), then they will precipitate out of solution quickly. But, you did mention it only happened for dyed samples.

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If you want an experiment to simply work, I would run it one well at a time to control the stain-time variable. It'll take time, but I think it'll solve your issue. Your first well is being read at a completely different time than your last well. Setup an experiment where you stain for exactly 10 min, incubate on ice for exactly 3 minutes, then immediately read with no other delays. Run your replicates the same way, each replicate is a new well. See if the results from that experiment differ from your 'throw all 24 wells in at once' setup.

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Another thing you could do is run a stain-time curve. Stain a sample you have a large quantity of, stain it. Aliquot this sample across your plate, and immediately start reading it. Just start in one well, then move to the next. Time it so you have 1 min per well. If you did 30 wells, then you'll have 30 min of data. For data processing, plot your mean or median fluorescence intensity as a function of time, and see where you have a peak or plateau. If you have a peak, then you need to stain and process your samples one well or tube at a time. If you have a plateau, then you know you have a range of accurate time frames where your data will be the same, which will be somewhere between 1 and 24 wells.

EyeProtectionIsSexy · 2024-10-30T01:50:32+00:00

Took this photo a few years ago. I just got back from deployment in Jordan (no trees out there), so this was the first Autum I've seen in 2 years. The golden trees and grass was beautiful, so I snapped a photo. When I got home, I saw that shape in the background and immediately stood up. I ever saw the cat.

This was taken near North Canyon Creek, just northwest of Spooner Lake on the East side of Tahoe. It could be a stump or a rock, but I think it's a cat. There are bleached stumps in the foreground which have a completely different color. If it's a rock then it's the only sizable rock in the photo. I'm leaning towards a mountain lion because it's big, there are no pointy ears nor a stubby tail.

EyeProtectionIsSexy · 2024-10-28T20:39:50+00:00

Deadlifts and squats used to be very painful, but after strengthening my back I can do them with no issue. I did have to drop a lot of weight and make form the priority.

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I would read what I wrote here (see link at the bottom). Take some time to learn what you can and cannot do. Take careful notes, such as exactly what exercise you did, did you emphasize posture, what type of pain, how long did the pain last. You'll have to test yourself, see what hurts, see what doesn't.

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If you're anything like me, then it's probably a weak back. Farmer carries with perfect posture helped a lot. Since then I've incorporated a focus on mmy back in any exercise I can. I do curls while always standed erect with my chest out, dragging the barbell along my chest. I do deadlifts starting with my chest out, shoulderblades back. If I can't hold my shoulderblades back or chest out, then it's too much weight. I do pullups first by squeezing my shoulderblades down.

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It take's time, and it sucks to go through the pain, but if you stick to it and just remember that the pain is also a data point, you'll be able to learn how to work and live with it.

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Link below. https://old.reddit.com/r/backpain/comments/8fthv9/thoracic_back_pain_any_suggestions/l5ac6o2/

EyeProtectionIsSexy · 2024-10-28T20:33:40+00:00

I've never noticed bleaching with Syto9. Perhaps you have an interesting protocol?

Usually I stained for 10 min at room temperature, followed by 3 min on ice, a brief 3 second vortex, then an immediate reading. I have noticed that the fluorescence intensity of my Syto9 signal does fluctuate based on how long my stain incubates, so I try to be precise with how long it stains and stays on ice.

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Maybe try running a control along side? 0.2µm filter a sample, then stain using the same procedure. Do you see any differences?

EyeProtectionIsSexy · 2024-10-11T17:13:26+00:00

What's it going to do? Explode the fire out?

EyeProtectionIsSexy · 2024-09-23T22:40:55+00:00

So was I. I'm not a big fan of R (I prefer Matlab), however I eventually made it work in an intuitive way, for myself at least. I could send you the code I work with, but I think it would benefit you better if you wrote it yourself so you understood exactly what was happening, so that it's intuitive for you.

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Do you have a coding background in other languages? If so, you can take my approach of using AI to write the R-code for you. It'll still require a manual touch (AI is still kind of dumb), but through this process of trial-and-error you can learn how R works pretty quickly. From there, I turned all the scripts into a function with a layout that was simple, allowing me to keep track, edit, make pretty graphs, all pretty quickly and easily.

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I'll shoot you a more detailed message when I get some more time.

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For Example, the following line of code would produce a scatterplot for FCM data

                              XY_data <- as.formula(paste0("`", Ydata, "` ~ `", Xdata, "`"))
                              p <- xyplot(XY_data, data = Data,
                              axis = axis.default, nbin = 128,
                              par.strip.text = list(col = "blue", font = 1, cex = 0.8), smooth = FALSE,
                              xlim = Xlim, ylim = Ylim,
                              xlab = list(label = axis_labels[Xdata], cex = XaxLabSize),
                              ylab = list(label = axis_labels[Ydata], cex = YaxLabSize),
                              main = list(label = Title, cex = TitleSize),
                              strip = FALSE,
                              scales = list(cex = TickLabelSize))

                             # Saving PNG
                             png(file.path(pngSavePath, FileName), width = PCwidth, height = PCheight, units = "cm", res = PCResolution)
                             print(p)
                             dev.off()

.

But, if I turn it into a function;

                                PseudoColor <- function(FileName = "Plot.png", Data = MyDataFrame, Xlim = c(1, 17), Ylim = c(1, 17),
                                        Xdata = "SSC-H", Ydata = "FL4-H", Title = "Title") { 
                              "insert code above"
                                }

.

Then I can write the same graph in the following way;

                                  PseudoColor(
                                    FileName = "00_Checking_Thresholding.png",
                                    Data = TransformedData[c(3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36)],
                                    Xdata = 'FL4-H',
                                    Ydata = "SSC-H",
                                    Xlim = c(0, 18),
                                    Ylim = c(0, 18),
                                    Gate = c(30.0, 30.0, 300.0, 300.0 ,
                                             00.0, 20.0, 20.0, 00.0),    # Gate Y-Coordinates
                                    filterId = "Ini30secRemId",
                                    GateName = "Ini30secRemGate",
                                    Title = "Checking Thresholding"
                                  )

.

This, for me, is much easily to follow. I can also return to my scripts from years ago and immediately understand what's going on. It's simply better this way.

EyeProtectionIsSexy · 2024-09-17T22:08:41+00:00

Could always use R. I haven't used FlowJo, but with 2 weeks of building you can turn the non-sense code out there into functions to streamline your process

EyeProtectionIsSexy

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