For those of you who work in a zebrafish lab, I'm aware that zebrafish like to mate in the morning and that's when we'll normally plan our microinjections/etc. However, it would be a big convenience for me if I could get them to mate later in the day, for multiple reasons (staggered mating means it's easier to reach an 18 hr time point, it might relieve equipment constraints, and also just let me sleep in). Does anyone have tricks to get zebrafish to mate later? I tried keeping their mating cages under a cardboard box to keep them asleep, but this doesn't always seem to work/only works for an hour or so.

hi everyone i'm a lab manager (21) who had an overlap of working with an undergrad (20) for 2 weeks before they left the lab. i have a crush on them but feel guilty about even entertaining it even tho we r sort of friends and they left the lab...

So long story short, I made tsb media in vials and autoclave them, and before inoculating them, we leave it in an incubator shaker to make sure they are not contaminated. Sadly they keep getting contaminated. I was wondering could it help to clean the vials, then autoclave them. Then make the media and autoclave it again to prevent this from happening ( contamination). Or is it too much or there is maybe something else happening that maybe my newby brain doesn’t understand or is doing wrong.

I'm not seeing it being discussed much here, so I'm curious if anyone knows about initiatives like The Freezer Challenge or the UN Race to Zero. If you do know, what is it like in your lab?

As the title says, I am constantly nervous or wake up in the middle of the night because I constantly think I left something on. This often ranges from thinking I forgot to turn off one of our microscopes to leaving our liquid nitrogen tank open. I've had dreams of coming into work and seeing our only tank bare dry and at room temp which contains all of our precious patient samples. Usually at end of day I will do a run down to make sure everything is turned off and double check before leaving for the day.

What are some things that keep you guys up at night?

Hi all,

I'm relatively new to cell culture and tried cell cell counting today on my own for the first time (I've been shown before and worked with someone as they did it, but I digress).

I'm culturing HEK293 cells and looked at the under the microscope and while some areas were bare, some were extremely high confluence and starting to dislodge (despite that I plated them from frozen Wednesday and changed the media Thursday). Therefore, I decided to passage them.

When I passaged them, I used an aspirator to remove media, added 5mL of PBS to wash and shook the plate in a cross motion. I then aspirated the PBS and added 2mL of trypsin/edta 0.025% and shook the plate in a similar motion before putting it in the incubator for 2 minutes. Following this I added 8mL of media to deactivate the trypsin. I then used a 10mL pipette to put the cells into a 15mL conical tube for cell counting. I used an automated cell counter and got a number of around 30k cells per mL. I wanted to freeze the cells and I know the standard is 1million cells, so I figured I may have done something wrong. I then centrifuged to get a cell pellet, aspirated the media and resuspended the cell pellet and re-counted. This time it was around 60k cells.. which again seemed off and extremely low.

I decided not to freeze the cells due to a low cell count and plated them 1:10 for the weekend.

Any and all help is appreciated.

Hey yall, I’m leaving for a somewhat remote trip to sample some fish. Going to be collecting fin clips for DNA sampling. Any idea what the best way to preserve DNA is in a pinch?

Will everclear work?

If I can get my hands on 95% ethanol with this be ok?

My lab does lot of perfusions and brain extractions in rats and mice. Our surgical tools scissors have really started to dull after a year of hard use. With multiple powered studies coming up we need good tools.

I’m looking to buy a new set for the lab, but have no idea where to start. Our current set probably isn’t worth being sent off for sharpening. Any suggestions?

Someone is looking for some cells from a guy who spent 5 yrs in our lab and left as a RAP.

His tube labeling is unintelligible, no dates, some gene name, no cell type name. But at least it's in black sharpie. The other postdoc labled everything in pink fine marker and it's fading fast. She crammed everything on the cap. Both of them kept either bad lists or no list at all. The newer postdoc has her own collection of 18 half full boxes and has to look thru each box to find things. The freezer is down to -69 and guess who has to stay late on Fri nite to make sure it's getting down to -80 again after she's had it open and i've been yelling at her to close it! Of course she too has no lists!!

I'm sooooo tired of this. I'm the lab manager who gets asked "do we have this, where is this, where is x's box located?" I tell every one of these people when they arrive to label their lab notebook with their name and their boxes with their name and if it's cells, plasmids, glycerols, protein, AND make a list, keep track of sources, cat#s, lot numbers even, but do they listen? most of the time they can't even cross of the name of the person who used to own the box 10 yrs ago!! Is this an archive or someone new?

Going into a box of someone who has just left the lab is also an adventure. Oh, there's that antibody we were looking for and bought again for $500. There's that chemical that was listed as being in another box altogether and the boss yelled at us about when we couldn't produce it. Aliquot! I yell! "Can I keep this antibody in my box if I am the only one using it now?" Noooooooo! Top Postdoc says I yell too much. Wait til you hear our PI yell, I say.

I'm stressed and totally underwater . . . We need to organize the -80, the -20s, the +4s, but every time I start, part way through someone's looking for something and stirs the box up, the boxes in the racks, the racks up! If they make the list and I read it off, I can't read their writing. And the boss' admonishment to not use up the plasmid, to keep 20 ul in each goes unheeded. Did you make a glycerol of this, I ask. Ummm, no. What did they do before when they were learning as grad students?

I'm going mad. I just had to vent. But at least the -80 is at -67 and it's not dark out yet. It'll be nice to get fresh air since the guy who just shut off the loud indian music has apparently left us breathing beta mercapto-ethanol and just missed me yelling about this, again.

Hey, next week, I start my first job in a lab at a private company after completing my PhD. I didn't lie during the recruitment process, but I'm still very nervous that they won't like me or that they'll expect me to know techniques I've never done before. I need your support. What was your first scientific job like after university?

Any pros and cons? Any tips to learn within a structural biology lab?

Hi, I’m an undergraduate student and a post doc researcher from Oxford offered me to stay 3 months in her lab. The only problem is that she can’t give me financial aid for the stay. Does anyone knows what international scholarships or foundations can help me to cover all.

Btw, I’m from Mexico and I’m an undergraduate student. Some scholarships are only for postdoc, PhD students or even graduate students.

We know there are a lot of things in the lab that shouldn’t be tasted because they’re bad for us. However, the lab tempts us with the allure of forbidden fruits. Has anyone tasted them?

I imagine DMEM tastes like fruit punch. In reality probably like fermented piss mixed with iron.

I imagine Dimethylaminobenzaldehyde tastes like Froot Loops.

What other forbidden fruits can you think of?

This has been frustrating me really strongly as of late.

I’ve worked in five labs total. Each time I went in for the interview, that’s when we’d discuss when I’d be joining. I was an undergrad so maybe that’s part of it. My understanding was that techs don’t know the techniques and you have to teach from scratch so onboarding is based on potential.

However. This lab I’ve applied to it’s been hell. It’s been six weeks so far and each week it’s something new. I’m just a Bachelor recent grad and it’s been intense. We did an interview -> 2nd technical interview -> refrences -> lab lunch. Each of these steps was 1-2 weeks long. Last recently, he said hey I’ll get back to you soon I’m interviewing a few more people. I’ve never had it be like this in my life or any lab.

I understand that different labs do things differently but this is bizzare right? Like it doesn’t take a month of interviewing and chatting to realize a candidate is a bad fit. Can someone help explain what’s going on, and if it’s just expected?

Hey guys, I am getting these crazy peaks since months. I’ve tried different primer concentrations and had a number of tries but I am curious to know what could have gone wrong ? Suggestions welcome

Does anyone have recommendations for a poster that has different metabolic pathways like glycolysis, pentose phosphate, krebs, etc?

That always gives me the motivation to power through school lol. I'm studying so that no more adorable cartoon sheep have to suffer

So, I need to visualize both Actin and Paxillin, and they are both anti-rabbit. But we only have secondary antibodies that are Goat anti- Rabbit, one Alexa Fluor 488 and the other 647? I'm guessing that won't work, though, right?

I'm thinking I need to use a Goat anti-Rabbit for one of them, and then a different host for the other?

Hi all! We are a smaller academic setting where I manage equipment requests by email. The only thing I'm really keeping tabs on right now are the laptop carts. Easy!

We are purchasing an anatomage table which will be shared with multiple departments. Vet tech, bio, nursing.

What system do you all use and recommend? I need to maintain some level of involvement as it will be the techs moving the table to the labs for the classes. Also, I would prefer we dont end up in a turf war with one department monopolizing reservations "in case" they want to use it. Any and all suggestions appreciated!

They're the tungsten carbide, microserrated, surgical scissors from FST. Retail for $220 USD. Sequins are a nightmare fabric to work with and you have to trim them all out of the seam allowance or you'll risk breaking needles when you sew over them. One of my previous positions the company shut our entire site down so there's was a lot of random stuff up for grabs.

So I recently took over as 'lab manager' for a small molecular genetics lab (so small it's just me now that my predecessor left). Context, I arrived in 2019, my predecessor had been here since 2008 and just left earlier this year.

I'm going through and organizing our -80 and -20 freezers and I am finding a ton of old reagents (polymerase, ligases, etc.) and competent cells that all have expiration dates ranging from like 2008 to 2015. A lot of it is no longer relevant to the work I am doing. So I'm torn on if I should keep it or not. It had always been drilled into me to not use expired reagents, but looking online I'm seeing mixed responses. As far as I know they have stayed in the freezer and were handled properly, but again it wasn't me working with them so I can't be 100% sure.

So thoughts, is it worth keeping reagents that expired 10+ years ago?

When I work in labs etc protocol is always to keep total quantity of flammable reagents to below 50L in the building at any given time.

Is it just me or is this pretty ridiculous? I presume the staff aren’t payed enough to know or care.

Of course some of them are acetone-free and others have been diluted with water. But the vast majority were acetone or ethyl acetate based. All with flammable GHS symbols.

On the bottom shelf the bottles were stacked 9 bottles deep. There are approx 20 rows of 9 on the bottom shelf. Each bottle on the bottom shelf was 150mL. That calculates to about 180 bottles @150mL on bottom shelf which is 27L. So the total in the picture should be well over 50L.

I can only imagine the stockroom as just as flammable.

Hi! I was wondering if anyone had any advice? I will be working on shaving mice for an atopic dermatitis model soon and I need to shave the right cheek, like on the face. A senior suggested an eyebrow razor since the area is so small, but I have zero experience shaving so I would like to know if anyone could help 🥹

I'm trying to determine to what capacity these machines, in general, can be used to help strengthen data authentication for a scientific pipeline.

Do you know about any machines that do this?

Are they typically included as metadata in their proprietary formats?

Anyone have any experience using this device for high molecular weight transfers? (~200-300 kDa)

just curious what settings/adjustments you make. Maybe target/tissue/cell line dependent but just looking if anyone has something that works very well. I tried the preset mixed/high molecular weight programs without much luck.

Hey I'm using the MagNA pure whole blood DNA extraction protocol and need my extracted DNA to be within a certain range. Obviously that's very hard to control. I'm using the entire plate from 1 source so I'm going to pool my elution into 1 tube. From there if I don't have a high enough concentration what can I do to precipitate out the DNA and pellet it to resuspend in molecular grade H2O?

I have reagents to perform manual extractions so lysis buffers, protein precipitation buffers and isopropanol. Would just adding isopropanol be enough to precipitate out the DNA and pellet it or do I need some kind of salt into solution to aid in precipitation?

I submitted an F31 to the National Eye Institute (NEI) in August. Study section met last week and I received my score, 19th percentile and impact score of 28, no summary statement yet. For things to do its flagged with a JIT and I can upload several things. NEI doesn't publish a payline, is anyone familar with F awrads at this institute and if this is a competitive score, or how I can find out? My PI is away, I messaged him but no reply.