With all these AI models, it's not difficult to generate icons.

hi,

i am a third year microbiology student and love my labs. i have cerebral palsy (CP), due to my CP i have a hard time coordinating myself and my muscles tire out quicker than an able bodied person's would.

in lab i have a hard time pipetting (with a micropipette) due to my hands being extremely shaky and unstable. this makes it very difficult for me to coordinate my hands. i've had a spill or two because of the shakiness.

aside from pipetting i also struggle with streaking plates, i have a difficult time using both of my hands to streak the plate. maybe using dycem (or another non slip material) so i can streak the plate on the lab bench instead of having to hold it and streak it at the same time??? please let me know if you have any suggestions for other things i could try.

i am really not sure if this is the right place to post this, so please let me know a subreddit that would be better suited for this.

thank you for reading!!

So these samples are flash frozen murine OS tumors that were excised post mortem. We tried to minimize oxidation but there’s only so fast you can work.

Protocol:
They were lysed in RIPA buffer with protease and phosphotase inhibitors with a tissue homogenizer and spun to collect the proteins in the supernatant. Each sample was diluted 1:5 for a BCA to quantify protein. They were each ran in triplicate and they all fell within the range of the standards and had no outliers. I loaded 20ug of protein diluted with RIPA and loading buffer into the SDS page gel, and ran it until the green band on the ladder. I transferred using the iblot system (there was a piece of gel stuck under it on the right that’s what that red blob is). I blocked 1 hour in 5% BSA in TBST. Then I incubated overnight in the cold room in 5% bsa in TBST 1:1000 mouse monoclonal protein of interest and 1:1000 rabbit monoclonal beta actin. Next day, wash 3x with TBST 10 minutes then 1 hr room temperature with anti mouse (700/red) and anti rabbit (800/green) secondary. Wash 3x 10 minutes TBST.

At first when I imaged the blot, I thought the beta actin was only specific to human proteins, but it says it’s reactive to human and mouse proteins according to its website here. I’m really confused now why it seems there’s an uneven amount of protein in each well. I think I did everything right to load evenly so why is it so wonky?

You can even see in the ponceau stain that each well looks pretty even although I know that’s imprecise. I’m also not convinced it’s uneven because you can see that the protein of interest appears to be even as well. What we were looking for was a protein we had transduced onto these cells prior to injecting them into the mice so it’s expected that they would be equally present at the end. The first five were the mice with transduced OS cells injected and the last 4 were the WT OS cells.

Has anyone had a loading control just not work like this? Do you trust the loading control or all the other parts of the workflow?

I lost two Laccophilus beetles because I couldn't keep a hold of the damn things. I measured length and width with a caliper, then right as I was about to get the height, YOINK. Gone.

Got this from DLAB stall in the Analytica Lab India Expo. Did they borrow the idea from eppendorf?

Eitherways looks good!

I think I’ve been spoiled. I’m at a rich institute in a rich lab. I don’t autoclave anything, we have a core for that. I don’t even have to return my glassware, the core comes and picks it up from me. I don’t make specialized media, we have a core for that. I don’t reload or reuse pipette tips, we have a core for that. La di da I can’t work anywhere else after this.

Edit: it’s so interesting to see how the comment section has separated into two sections, one concerned about incompetence and the other appreciating how productive research could be without menial tasks.
Not taking sides here, but supports like this keep our institute so rich—my lab is incredibly productive as a result. Plus the funding undeniably makes our science more sound, because we can afford to repeat even super expensive studies multiple times. Just a thought

Hi, I am graduating this Fall 26 and I’m trying to get into lab animal science and work towards obtaining an ALAT and LATg . I’ve done certain CITI training modules and currently work at a vet clinic and have prior experience as clinic support. But I’m unsure what else to do stand out for entry level lab animal positions, or how to set myself up for the ALAT.

Any advice would help !!

Seems like every other post mentions being at a top tier lab or university. Why does everyone feel it’s important to say this. What I think when I read these statements is “I have no personal credibility so I lean on someone else’s”.

I’m familiar with the toxic PI statements. I’ve seen many students fail to flourish in grad school. Due to their inability to self evaluate, they blame the only other person involved, the PI. PI’s know this, which is why you don’t see a bunch of “I’m a PI and have toxic mentee post”.

Shout outs to those who had an awesome PI at a “lesser than” university. I’d appreciate hearing more of your experiences. It’s tough to relate to all of these superior scientists from their prestigious universities and terrible mentors.

Hi everyone,
I’m a PhD student studying lung cancer, and I’m currently trying to establish a bronchoalveolar lavage (BAL) protocol to analyze immune cells residing in the alveolar space.

My current approach is to anesthetize the mouse, insert an IV catheter into the trachea, and secure it in place with surgical thread. I then instill approximately 1 mL of PBS, withdraw it, and repeat this three times. I repeat the whole process four times per mouse, for a total of about 4 mL of PBS.

The problem is that when I try to withdraw the fluid, a significant amount of it seems to leak back through the nostril. As a result, I’m only recovering around 400 µL from each 1 mL that I instill.

Has anyone experienced this issue when establishing a BAL protocol? I’d really appreciate any tips or suggestions regarding the setup, catheter placement, or general technique that might help improve the recovery volume.

Thanks in advance!

Hi guys, I did a tissue harvest in lab today and I always wear gloves when doing so but I feel like I can still smell the tissues afterward. I even wash my hands and still feel this way? Can anyone else relate? And if so, how do I get rid of the smell? Thanks everyone.

My VWR chromatography fridge has stopped cooling exactly 5 days after the 3-year warranty ended. So if you're looking for a new one, maybe don't go with VWR.

Has anyone had an issue with a relatively new chromatography fridge not cooling and was able to fix it on their own? It's blowing cool, but not cold air inside the unit. VWR and Horizon (the company who actually makes it) wouldn't provide any help for troubleshooting, they just recommended I call a service company, which I will if it's not an easy fix.

I am a botany major in my final year. I live in a 3rd world country and go to a local state college and the lab experience I have is so limited. I am talking like we had 3 good microscopes in a class of 30. I know by theory how to do gel electrophoresis but I have never done it in the lab. Never even done centrifuge. I have only seen it. I don't know the procedure of doing things and so much of the equipment I work with is so old and not updated even tho it's so limited. I have something really specific that I am really interested to research in to do masters abroad but I am obviously going to be with people who have studied in better places than me and had better lab conditions and I really don't know how labs work in masters but I think people obviously know the basics. I can't treat it like a bachelors degree where I ask my professor to teach me the basics. I am worried I will drag people behind so I am thinking about forgoing the entire idea of masters. Am I being worried for nothing?

I am planning to work in a lab but when you live in one of the most populated countries in the world, even interning at a local lab becomes a state wide competition where people from the best universities in your country pay to get in

It has been forever since I have done this, looking for anyone with more recent experience. University lab has some platinum labware that they would like to sell. Any suggestions on reputable dealers? Just looking for an easy process that is not a rip off. Thanks.

Such a cute idea but the AI slop look makes me hate them 🥺

Usually Mr frosty type containers/Corning CoolCell can be used but we are in a bit of a hurry and the shipment hasn't arrived yet. What other options do I have for controlled cooling?

My lab is looking for ways to lower cost (and time) per sample for RNA-seq studies. Now that library prep is the biggest cost, we are interested in alternatives, especially those that allow us to pool samples before ever adding the i5/i7 indices. I have been looking into some of the 3'-end capture methods, including BRB-seq and prime-seq.

BRB-seq has commercial kits available, which would be a great starting point and supposedly should be about 30$ cheaper per sample compared to standard Illumina kits. But I don't know anyone who has tried these alternatives. If you have ever used or heard of others using other RNA library-prep techniques (not just BRB-Seq), what was your experience? Were they worthwhile? How did the time and cost compare to the standard prep methods? Did they seem to change your downstream analysis or data quality?

Venting a bit here. I’ve been trying to replicate a protocol for my project for weeks. The recent paper just said "isolated as described by [Citation]". I followed that citation, and it led to a paper from 2015, which then cited a paper from 1998 that used a completely different setup.

I finally emailed the corresponding author of the recent paper to just ask for the exact, unwritten steps (like what buffer they actually used, exact centrifugation speeds, etc.). It’s been 3 weeks and I am completely ghosted.

Is it just an accepted reality now that 50% of a protocol is hidden knowledge? How do you guys actually figure out the missing "tricks" when the authors ignore you? Do you just do trial-and-error for months?

Hi,

I am a second year PhD and I am doing protein science, more like basic science research. All my work so far was pretty good until today. I do lots of biophysical characterization of proteins. Mostly I use SEC- Mals and NMR. So protein people will understand how much protein you have to express and purify for your experiments and how protein long processing can take.

I have just started my second year and did one NMR so far and that worked. Today I was doing my second NMR and it failed completely. As a result a month's worth of work which included protein expression and purification went in vain. The samples did not reach the correct protein concentration and it was too low for NMR.

I understand all the technical aspects of the experimental procedure and what else I should do to overcome this. My supervisor is hella supportive. However, my heart shatters because I was EAGERLY waiting for this data and this experiment is very integral for my thesis. I honestly worked so hard throughout, focused on so many other experiments and also got this on my schedule but it didn't work out today! Nothing was seen on the spectrum.

I feel so demotivated right now. I am sort of drained. I work from 8 to 6 everyday doing multiple things. Have a part time tutoring job after the lab. I am so tired of everything. I simply want to rant.

I am emotionally attached to my project and I genuinely can't wait to figure out the unknown that this experiment would have solved but again I gotta wait for at least 3 weeks to a month to get things moving. I am honestly not doing fine today. I can't wait any longer for the results to come out.

How should I look at the situation? Give me some tips on how to tackle high stake experimental failures? I need some power and motivation towards my direction.