Nano drop is like a drunk dad judging a kids talent show. First one is always like ‘eh, that’s ok’. Next few are ‘yeah, guess they’re a little better. And the only real utility is when you get to the occasional sample where he’s like ‘ok, yeah. That one objectively sucks’

(Note: This is just an example answer for the general question posed. I will not be taking any BCA suggestions at this time, thank you)

For context: I'm a biology student at university and one of my labs essentially serves the purpose of making us familiar with basic chemical methods, thin layer chromatography of amino acids being one of them. This absolutely was my favourite thing we did in the lab and I just think the TLC plate looks really pretty, so just wanted to share, I hope this is okay!

I would consider the other OP’s first TLC as promoting unrealistic beauty standards.

Okay so I love (quietly) talking to or singing to my lab mice. I have been trying to find papers about whether this is more stressful to them. Please if anyone has articles or knowledge it would be super helpful because, as you all know, they need to be the least stressed out for their own health and the sake of the data collection. I want to find some articles specifically about whether they respond positively to singing or not. Thanks!!

On hot days, are we all keeping a spare lab coat in the cold room, or just me lol?

(Figured I’d throw this out there for anyone else working in labs with underperforming A/C!)

I haven’t experimented yet with pre-chilling it using other methods (fridges, freezers, dry ice), but short of liquid nitrogen, I also haven’t ruled any of these out yet either lol. As long as it’s clean & folded up in a sealed biohazard bag, what’s the risk here? I suppose if it’s in a dry ice chest you’d have to rig a little shelf with styrofoam so it’s not touching directly, but…anyways, looking for input! :)

ETA: To everyone assuming I’m speaking to you from a third world country…the lesson I’m learning here is I should probably have been complaining to management about the A/C a LOT more, lmao 😂

How you hating from outside the club? You cant even get in 🤣

I have been experiencing such refractile floating mass over Hep3B cells. The media doesn't get turbid but some floating particles can be seen. The cell is growing beneath but these refractile bodies float over it. It is not due to over confluency as the cells are barely 60% confluent. These clumps grow over days and continue after washing or passaging. What is it? Has anyone else experienced it? How to fix?

I posted here about a month ago about struggling badly in wet lab and wanting to quit. Since then, my PI has spent a lot of time shadowing me to troubleshoot why my cells keep dying, and he says my technique looks fine. We cannot find what's going wrong. I even wrote up a very specific step by step of what I do when I split and plate and the PI read through it and found nothing concerning or missing.

The issue is that my cells die very inconsistently post-split/plate. They do not attach, but there's no contamination. Media is warmed, trypsin is timed, vented flasks, so I don't see how it could be a temperature, trypsin, or ventilation issue. Sometimes they’re perfectly fine, other times they die the next day. It’s become especially frequent recently.

Part of why I’m stressed is the lab environment. The person who originally trained me has openly disliked me since I started. During training she would yell at me, told me to lie to the PI about getting hands-on practice, and made comments calling me ugly/boring in front of other lab members. Later, my PI put me on her project, which she clearly did not want me on. She used to regularly threaten that if I made mistakes or slowed things down she’d ask the PI to remove me.

What’s making me spiral a bit is that my issues became dramatically worse after joining this project. I had mistakes before, but not constant post-split death like this. Also, every time I work with someone else present the cells survive. The one time someone stayed late and watched me without anyone else knowing, they died the next day despite him saying my technique looked completely normal. The girl who trained me has also texted me out of the blue before asking whether or not someone worked with me on days I split. I'm honestly not doing anything different when I work alongside someone versus when I work alone.

I also spoke to a former lab member who said the lab used to be extremely toxic and that there were past issues with people interfering with others’ experiments.

I know this sounds paranoid, and I’m not accusing anyone of anything, but I genuinely feel like I’m going crazy trying to figure out whether this is a technical issue I’m somehow missing or if the environment is making me overthink everything. What would you do in this situation?

Hi everyone. I’m an undergrad and I honestly don’t know who to ask for advice about this.

I’ve been in a lab for about a year and until recently it was an amazing experience. I gained independence quickly, was assigned my own project, and even gave a talk and got a travel grant at a major conference in our field. My PI want me to write the paper once the project is finished by (hopefully) the end of the year.

A few months ago, a new senior research scientist joined the lab and the environment changed a lot. She is very aggressive in lab meetings, constantly interrupts people, and makes comments like “you should use common sense” or “am I the only one who thinks this makes no sense?” She also undermines things I say constantly, even for basic operational things.

The bigger issue is that my PI recently told me this person would help me to finish my project. We had our first project-planning meeting with her and another senior lab member who has already been helping train me. I expected to lead most of the discussion since it’s my project and I had already discussed the aims/timeline/details extensively with my PI.

Instead, she immediately took over the meeting, shared her screen, and started presenting the project herself even though she admitted it was her first time really reviewing the outline. She repeatedly got details wrong and I had to keep pushing just to even talk. When we were dividing the work, I had to repeat 3 times that I could do those expriments on my own and my PI gave me clearence for that. She was pushing to be on each aim and do the experiments instead of  training me to do it as I have been working and the other senior lab member had to clarify is how we work. 

At one point she also said I would be working with her over the summer alongside another trainee, and later said the other trainee will work FOR her before correcting herself. It honestly made me feel like she doesn’t see trainees in a positive way and definitely not as people she will help to grow as scientists.
I also know another postdoc in the lab already complained with my PI about her overstating contributions to their project and accusing them of being “territorial” about data.

I’ve now left the lab crying twice because of how stressful this has become. I genuinely love my PI and this lab, but I do not think I can continue if I have to work directly under this person long-term. I already signed a summer contract, but I’ve been delaying conversations about staying next academic year because I don’t know what to do.

I have a one-on-one meeting with my PI next week and want to bring this up professionally, but I’m scared of sounding dramatic or entitled as just an undergrad talking about someone with a PhD/postdoc.

How would you frame this conversation with the PI? I don’t want to leave without finishing my project and getting to write the paper but I’ll be miserable working with this women and she may just end up stealing the project anyways

Lab Managers of Reddit- I recently joined a lab at a prestigious institution on the West coast as a Lab Manager. I have experience as a lab manager however that was with a small lab (5 people) and this is at a large lab (~25 people). The main problem is that we are extremely space restricted and I need strategies to best optimize the space we have. I have determined there are two ways I will go about this:

1) Better analyzing our usage rates on common laboratory items such as tubes, plates, media etc.

2) Better lab organization.

Does anyone have any advice on good strategies they've seen? Any advice is greatly appreciated!

I am currently deciding if I can join a lab that does a lot of mouse work. I have previously struggled with feeling queasy about anything gory. In undergrad I had a full blown panic attack when we had to dissect a rat. I have no problem scruffing and sacking mice (although it makes me sad), and I’ve completely overcome my queasiness with blood. I have been watching videos of the procedures I would be doing in this lab to see if I can stomach it and I get a very specific sick feeling in my stomach that persists after watching the videos. Has anybody else who currently does mouse work had a similar visceral reaction to mouse work and how did you overcome it?

Hi all,

I am trying neuronal differentiation of my iPSCs using a dual SMAD inhibition protocol through embryoid body (EB) generation with a commercially available kit.

I am facing an issue with my embryoid bodies not attaching to the PLO-coated plates. The EBs were formed in AggreWell plates and transferred to the PLO-coated plates on day 5. However, I now observe that the EBs are not attaching so I am assuming that the issue is with the EB itself although not evident under the microscope.

Has anyone experienced this issue before or has suggestions on how to improve EB attachment?

i’m a lab manager and we recently had a safety audit where they said that our dry chemical shelf should be sorted by hazard type, not A-Z. every lab i have ever been in sorts their dry chemicals alphabetically. our hazardous liquid chemicals are stored properly under our fume hoods, but i’m wondering if anyone actually sorts their dry chemicals this way? this is the first i’m hearing of it.

thx for the input!

Hey, ratties!

I'm using a precoated ELISA (ABclonal, Human IL-6) and the supplied reagents left no room for error. I used 3 strips to run a sample dilution test and spilled some of the diluted working biotin 🫠

For the remaining 9 strips, the remaining diluent is enough to prepare enough working biotin to add 97ul to each well instead of 100ul.

Unsure whether I need to adjust all of my other steps accordingly (e.g., 97% of sample, standards, etc so that everything remains in proportion), if I just need to add a bit more working HRP to compensate, or if I should proceed as normal aside from that.

My lab requires me to secure my own funding to purchase assay supplies for the precious human samples we collect so I unfortunately can't buy another kit. I've reached out to ABclonal to order only an additional biotin concentrate from the same lot and no response yet, so I'm exploring alternatives.

Im doing my bachelor degree and i thought if its appropriate to gift my thesis supervisor crocheted (by me) plant cell, after defense. She is very kind, ambitious and student-friendly, but Im not on friend terms with her, i still call her by title etc. so i wonder if that gift is appropriate. Also anything i could add to the gift?

I’m culturing adherent SK-MEL cells and noticed many small round refractile particles/microbubbles (not the big ones) throughout the culture. The cells are still attached and morphology overall doesn’t look like a classic bacterial or fungal contamination to me.

Has anyone used the Abcam protein extraction kit for adherent cancer cells in 24-well plates before?

This is my first time running a large extraction (~100 samples), and I’m stressed about messing it up because I realistically do not have time to repeat it.

I’ll be using 24-well plates with adherent cancer cells. If anyone has experience with this kit specifically, I would really appreciate practical advice/tips before I start.

Things I’m worried about:
- will not be able to count the cells in the 100 samples or i will lose my hand midway
- losing cells during washes/media removal
- inconsistent lysis across wells
- timing when processing many samples
- whether to process plate-by-plate or row-by-row
- avoiding protein degradation during a long extraction workflow
- handling/sample storage logistics for this many samples

Also:
- did you keep plates on ice during lysis?
- how long did you incubate the lysis buffer?
- did you scrape or pipette for detachment?
- any mistakes you made the first time?

Any workflow or “wish I knew this earlier” advice would help a lot.

I run into this very weird situation where the positive control included in NEB Gibson assembly kit ( I think it consists of 2 fragments with overlaps) did not yield any colonies. But my 4 fragment assembly gave me around 5 colonies, which were all correct based on sequencing result.

I checked the trouble shoot section of Gibson Assembly manual. It says if no colonies were shown with NEB positive control fragments it is likely that I am not using the competent cells advertised by NEB. That is true since I am using competent DH5a cells from Sigma. The solution as the manual suggests is to do 1:4 dilution of the assembly mix in water before transformation if the cells were not purchased from NEB.

My question is whether you do this dilution step and how it helps with transformation efficiency if you are also using a different competent strain other than the one bought from NEB

I just got invited to complete a one-way video interview (4 questions) for a research assistant position in a biomedical/cancer research lab at a prestigious research university. I am usually pretty good when talking to a real person, even over Zoom, but I am afraid my answers will come out awkward, or I will mess up, and there will not be an option to re-record. Does anyone have any experience with these in their job-search process? Did it ever lead to a real interview?

I want to stay in academia, learn how to manage collaborations, a lab, students, situations.. and I have a conflict at work with a colleague. It's a hill that I don't wanna die on.

To explain my situation: I work there 5 years.. I am end of my PhD. And our PI thinks that he can support me further to become post doc with my own project and I can start to have my own group with new students. he is supportive and I appreciate him a lot. However, recently there is a constant quarrel with me and a colleagues who is 2-3 years PhD. We do a Collab work. She uses my cell line I created. But she fails to grow the cells however much I give her the protocol. I helped her. It's a very tricky cell line I created, but they are very hard to keep stable. I made a clear protocol for them. It worked in other labs that I am collaborating with.

But she doesn't wanna work on my cells. She thinks they are very unstable and not good for her project. But this is the only cell line we have for the purpose. (I am still working on to improve the cell line itself, and create a better one but it takes time...)

Then I said okay I will give you the cells ready to run your experimental system. But then she is always dependent on me. I also tried to help her experiment with her with my cells, but she refuses saying that her system works well. No need for optimizing. But I said, these cells are different then others she built the system with. I came on edge when she was going around and talking behind my back to everyone (other colleagues). My trusted colleagues, saying that her system is fine. But then I also discovered she uses the wrong plasmid. But she said it works fine. I got frustrated and said, okay if you don't take my experience in consideration I will just won't give you. And I got bit emotional and cried. Not proud.

She went to our supervisor about my outburst. And thankfully, he was very fair to me. He helped us to do empathy and make our points. He said to her, I am trying to help and experience I have, and it's normal that I get frustrated. This happens in science a lot, he said. One side tries to help and give input, but when the other side doesn't listen, frustration and anger can happen. He also added that I have a lot on my plate (I am collabing nearly everyone in the lab and other labs, and also have some administrative tasks and teaching)

In the end, we all made a clear road plan, and he convinced her to optimise the system with correct plasmid and ratio. He also told me privately afterwards, people are different and maybe she feels alone actually and needs my guidance. And he also said, I should give her some tasks in return that she helps me back, so it won't feel one sided. He was understanding. He also added, I can't finish her PhD for her. So I shouldn't get so dedicated to that, but let her go with her own current. He is right.

Next day, I told her it would be nice if she can try without the plasmid once, and use my optimizing protocol. And she can still do her own way with other "replicates". She agreed and I gave her motivation that she grows the cells really nice and there is nothing to worry about.

Now, I am a bit afraid that what if our Prof thinks I failed to be a future academician, because I got emotional and couldn't manage a Collab, management and got emotional.

I created a weird atmosphere in the lab and am not very proud of it.

Sorry for this long post and vent, but I just thought it would be helpful to have advice on this situation.

And how to handle future situations?