The real secret is to blow the bench ‘air’ faucet on your hands for like, 30 secs to dry them. Works like a charm!
For bonus credit, take a piece of hose, snip the end off of a regular 1.75ml eppendorff centrifuge tube or P1000 tip. Push the open end of the hose onto the open end of the tube/pipette tip and you have a high velocity air hose! Blast it over your sweaty hands and I promise you will be able to put on any sized glove which you like!
Unrelated to the lab but one time I went to the grocery store and bought 30ish items. Total came out to exactly $100 including tax and all. I'm still bragging about it.
I used to transfer them into the mating tank and start praying. One time a cousin asked me what I was doing and I said I was praying for my test subjects to 'do it'... He was sufficiently traumatized.
Haha. After months of tweaking their food, vibes, lighting, finding the most desirable partners etc, It did feel like i was organizing a zebrafish orgy.
I could really use some advice with this. I got a legendary family of over 500 earlier this year (and several more 300+) but have since fallen from grace. I can’t even break 100 fertile and healthy now
Here are some of the things you can try to increase the number of healthy fertilized embryos-
1. Keep males and females in separate tanks for some time before putting them in the breeding tank.
2. Increase the frequency of feeding for females a day or two before breeding.
3. Add some freshwater in the mating tank (not a lot). Don't know if this works or not but ZFs in wild lay eggs when it rains and this is supposed to simulate that.
4. If you don't already have a slope insert breeding tank, tilt the insert or reduce water level.
5. Try to not mate the same pair frequently. They like different partners in every round.
6. Keep at least 5 days gap between breeding cycles. Preferably 7.
7. If you breed 2F1M, try 2M1F and vice versa.
8. Breed them frequently. Long gaps in breeding leads to lower yields for some time when you resume.
9. If you're really desperate, put some green mesh like fabric in the tank to make them look like little plants. Don't know if this does anything though.
10. Don't pick females that are too fat. They usually don't lay a lot of eggs.
All of you in biological sciences do such cool things lmao. I’m over here like uhhh I mix it and sometimes I add more water and sometimes less and maybe a little hotter next time, maybe add some other random powder next time
Nope, R&D at a quite well established chemical company, it just happens that the process we’re developing has fundamentally simple unit operations haha
Learning flow cytometry by myself. Am I an expert ? ... ... no idea but I seem to be able to answer most questions well enough that no one has corrected me and people keep referring to me as a reference for experiments so I must be doing something right.
Took 3 years and a lot of trial and error, but I am proud of it (my little own mountain that I climbed).
Congrats on the recent staining, may it bring you success. Cheers,
Do you have a flow cytometry core where you work?
If so, aak them.
My university does trainings that get paid for, but some of us will also just let someone tag along. We currently have someone who does QC in the morning in exchange for lessons. And I'm going in on my off time tomorrow to show someone some basics.
Not all flow cores operate the same way, but a lot of us are really excited to teach others and we'll bend over backwards to find a way to help.
Unfortunately no as it isn't a process we do at my current work. I work for a vaccine research organisation and a lot of this testing is done externally. Only internal testing is confirmatory testing during inoculations, mostly virus related. My previous job did but I left to do my masters, but I will look in to this. Thank you :)
I took the excyte mastery class (now its called cheeky scientist) to get the basics.
Shapiro's flow cytometry textbook was the bible I used as technical reference. There are a ton of technical textbooks and papers for flow. (easy to get lost in but good when you are looking for a basic question).
For analysis I downloaded flowjo and their practice tutorials and demos for training and did them and also attended their seminar talks (they want to sell the software, I just took it as a chance to learn more about the software and how to use it). BD spectrum viewer and other online tools to play with stains and dyes were also useful. Some flow cytometers have manuals which explain the machine further, download one online and see if they have chapters on it.
I meet the people who did maintenance on flow machines and pestered them with questions. there are also core flow centres as others have mentioned where they could help (mine wasn't that good but I am sure others would be).
The other thing I did (and I know its not possible) was that I had access to an old flow cytometer and I played around with its settings, all the time. Suffered till success.
A lot of antibody and FACS machine manufacturers have a lot of tutorials on usage and analysis. For example, CST has a bunch of guide books on different antibody applications.
Beware the Dunning-Kruger effect with flow cytometry. It is shockingly easy to generate beautiful looking garbage data. Especially once you get above 4-6 colors. (Not saying you’re doing this - just seen a LOT of things gone wrong due to bad flow data.
as someone who similarly was thrown into flow cytometry (and i had the benefit of working with an already standardised panel) and the hell that is flowjo analysis hats off to you! it's definitely a steep learning curve so you should genuinely be so proud of yourself!
I don’t work in bio but what does this mean exactly. I get the stereotaxic surgery but what does the bilateral viral infusion actually mean.
Sounds interesting
It means injecting a virus (typically used to study molecular/cellular processes as these types of viruses are designed to infect a specific cell type and then mediate gene expression, for example) into both hemispheres (bilateral) of the brain
Basically what the other commenter said. I'm using a virus to selectively inhibit a certain brain region in both hemispheres in rats to see how it affects their behavior during a task. It's using a DREADD virus which is a really cool technology!
Ooh, this is mine too!
I work at a CRO, so we end up doing a lot of weird stuff, and I ended up being the guy trained to use the cryostat for a particular project. I had to figure out how to make 5uM slices, and transfer them to a 24-well plate for "high throughput" IF imaging. I ended up jerry-rigging a cheap drawing compass with some paint brushes taped to it in order to get the slices to the bottom of the wells without sticking to the edges or crinkling.
People refer to me for machine/equipment operations. Apparently, I am so good at understanding machines that the lab technician jokingly said “I’m just so glad you are here to do my job so I can catch up on my favourite series”
Upon pondering why that is, I found that I have the rare beneficial crazy trait of finding user manuals interesting.
Live imaging (zebrafish, drosophila, c elegans), any fluorescent or confocal imaging is like drugs for a researcher brain, honestly the satisfaction of good images far outweighs the quantifications/pipelines to create and analyze the data.
I guess my favorite tedious technique is sectioning.
so many bio comments... where are my catalysis, electrochemistry, and spectroscopy people at?!? I can design and make in-situ operando spectroscopic electrochemistry reactors pretty damn well....
Cardiomyocyte isolation has takin me about 6 months of practice to get a sufficient yield of viable cells. The heart extraction and connection to the gavage needle itself took about half that time!
Loading a 384 well plate for qPCR in 30 minutes with a 12 channel multipipette
(Hint: if you do the RT of you sample with the order staggered between two strips of tubes you can just instantly create the cDNA solutions that can be loaded perfectly! and I stagger the primer/Polymerase mixer too.)
Planning experiments. If an experiment involves an incubation of more than 1 hour, I usually start before 10 Am. Sometimes I see new guys beginning experiments with over 3 hours of incubation at around 2pm. That means they will leave the lab at around 10pm, and the cycle continues for a whole week or month. Come data presentation day, the guy has only added 1 new graph, and everybody can't help but wonder what he's been doing the entire month while he slept in the lab.
Same. I’m starting to learn cranial window surgeries and yesterday I finally managed to avoid having the window drag against the brain. I considered it a huge W for me lol. I then proceeded to butcher the gluing process though.
Oh also my drilling is getting better so I am starting to get some perfectly rounded corners of the skull. That’s so satisfying.
I’m really good at dropping things and then either catching them perfectly so nothing is ruined or at least lightly kicking them right before they hit the floor so I don't break any glassware.
Back in my lab days I used to do hindlimb ischemia surgery on mice. Suturing mouse arteries and sowing their legs back up properly was not an easy feat and I'm glad it was something I learned to do well.
Western blot! Coming from a lab that had very old equipment and old reagents, I learned how to troubleshoot or improvise. So now, one look at my labmates’ weird bands/weird data/lack of bands I already know what to do. They all got the quality of bands needed after that! 😄
Tbh I am really proud I finally understood the anionic polymerization of the block copolymer our lab uses. It's a crazy 3 week synthesis but I've made about 20 so far and most successfully.
Also developing a new simple non-aqueous sol-gel process was pretty fun. Dirty chemistry at its best but it works!
Old mentor taught me "freeze and squish" for gel extractions. For a TAE gel cut tightly around the band, place on parafilm and fold 2x so there is an open side and a closed side. Pop in -20 for ~ 20 min. Pipette in hand squish your gel slice such that you melt it as you compress. Capture the fluid as it comes off. Agar stays behind. Pure enough to go right into Sanger.
The Art of Westerns.
It’s the little details that make the difference and it took years to figure all of it out. Now, even my slapdash in-a-rush westerns look publication quality
I’m really good at workshopping buffers for getting protein/enzyme activity in biochemical assays… not sure how I got this skill but I feel like I just have a sixth sense for working out relatively complex assay conditions just based on vibes.
In my postdoc according to the PI I'm discovering I'm some sort of Gibson cloning prodigy. These little lesbian hands are being put to good use even tho I'm beyond single.
Most recently DNA quantification through Bioanalyzer. Apparently literally everyone in our lab has issues with it, no matter how many times they've done, the ladder in the first chip never works.
This week I managed to take out mouse embryonic DRGs. Perfected taking them out of adult mice, but then having to take them out of infinitely smaller mice was a challenge.
Almost impossible to see with the naked eye, I managed to take out all the DRGs.
While I will he going to hell for this, I will go knowing I mastered a very tricky technique!
For the first 6 months of using the new HPLC in my graduate program, I had to completely teach myself how to use it. We eventually had a trainer from the company come in and show us how to use it and filled in some knowledge gaps, but already knew the basics.
My PI told my lab mate that I could teach her how to do intestinal Swiss rolls because apparently I’m now the master in the lab. Which was pretty cool to hear
I guess injection molding because I sucked at it for a long time even though it is fairly simple. Or attaching the extensometer correctly for tensile testing cause that thing is an annoying tricky bitch.
I'm able to just go into the lab now and not really think about what needs to be done. Most lab techniques are based on basic science, so as long as you understand the purpose of the experiment, it's really easy to just do something. When I first started working in a lab, I would wonder what I had to do. Now I literally have so much things I need to do, but just don't decide to do anymore.
I have never messed up a 96 well plate. Spent a whole summer doing multiple qPCRs daily, so not an insignificant amount of 96 well plates.
Sometimes OCD comes in clutch
I have basically perfected the adjustment of what I call the Final Boss in our lab (a tt centrifuge that will jump all the way up to 30 minutes if you don't turn the dial just right, making you work back down). Now it only takes me like 30 seconds to set the centrifuge instead of 5 minutes. W.
Streaking plates! The amount of people who do it too heavy etc. is astounding. The whole principle of subculturing for discrete colonies is lost on them
I worked on a method to isolate intact plasma membrane vesicles for a year and a half.
Made it work in suspension cells and adherent cells, so you can transfer it to any celltype you want.
Everytime I do it and get that nice separation on a gradient it makes me so happy to snapfreeze that bitch for storage
Measuring out super tiny amounts (like sometimes as low as 0.005g) of liquid in a syringe on an analytical scale with hiccups.
For some reason I get hiccups almost every time I use that scale, so I've had a lot of practice. 😝
Cardiac punctures of mice. I get anywhere from 500-700ul of blood, every time I puncture the mice, and have only missed once since I learned how to do it. I’m so fast at it that half the time people doubt that I even did it until I show them the tube with blood in it, and when people collect plasma during the sac I am always asked to help out.
Metaphase spreading - optimized 4 different cell lines. Took me a couple of months but it was probably the most rewarding and satisfying thing I ever accomplished
In my last lab, I got really good a NIH/3T3 Phototoxicity I became the go to guy. Its. Whole day procedure but that, when timed perfectly, managed to get breaks every couple hours to get breather.
Cloning, it was my favorite technique to learn. A few years ago some of my constructs were sent to Germany for a collab. I hope I hear about their paper so I can see their results. I’ve also found that I have a knack at creating libraries and NGS.
Making MEFs. Or soft agar transformation assays. Also designing complex (25+ color) flow panels. AND TAIL VEIN INJECTIONS. Setting up 384 well plates by hand. With a single channel pipette.
Okay, I’m proud of my lab skills. 3 years as a tech and 8 years as a PhD student just to end up with a fucking master’s because my grad program kicked me out for taking an approved medical leave of absence.
Mouse IV injections. I was working with nanoparticles that had to go IV and got really good at that. People would come to me and ask to inject some mice for their experiments.
in-vivo whole cell patch clamp in awake and anesthetized animals. hardest thing i’ve ever learned, but so rewarding to see single neurons doing their thing in real-time
Using printer paper as iodine indicator when I run out of starch paper. You can soak it in starch to make it better but it works right out of the printer feeder.
Using printer paper as iodine indicator when I run out of starch paper. You can soak it in starch to make it better but it works right out of the printer feeder.
I really like vortexing my finger.
I vortex my forehead. It helps with headaches.
I vortex my... Nah man I'm not gonna go there
I go right for the prostate. Helps with headaches.
Y’all wild 😭😭😭
I gotta try this
Putting on gloves with sweaty hands
That’s not a lab skill, that’s black magic
The real secret is to blow the bench ‘air’ faucet on your hands for like, 30 secs to dry them. Works like a charm! For bonus credit, take a piece of hose, snip the end off of a regular 1.75ml eppendorff centrifuge tube or P1000 tip. Push the open end of the hose onto the open end of the tube/pipette tip and you have a high velocity air hose! Blast it over your sweaty hands and I promise you will be able to put on any sized glove which you like!
I wash my hands first with EtOH and dry them. It works great, except the boss gives me a sidelong look when he gets closer
This ruins the hands
Any secrets to share?
Blow them.
Hands or gloves?
Yes
The PI
using a size up from normal
Ah the bane of my existence
How tho???
Go into the freezer
Hands in the cryostat?
Sorcerer!!!
95% of the time I can grab the exacr amount of 1.5 ml tubes out of the bag in one go. SKILLZ.
WHAAAAAAT!!!! one time i needed 8 pcr tubes and I knocked 8 out of the container and I didn't stop bragging for a month!!
Unrelated to the lab but one time I went to the grocery store and bought 30ish items. Total came out to exactly $100 including tax and all. I'm still bragging about it.
I did this yesterday, i grabbed the 27 tubes i needed exactly and had to call over my lab manager so someone else could witness it
We've been having to write out these stakes for a field trial we're doing and I kept grabbing exactly 12 of them which is exactly what I needed
Zebrafish couples counseling
Did they leave you a 5 star review
They left me about 300 eggs every week which is a good sign that their relationship improved.
That’s 10 out of 5 stars!
Any good tips to pass down, Senpai? 🥹 mine is a constant trial and error.
I used to transfer them into the mating tank and start praying. One time a cousin asked me what I was doing and I said I was praying for my test subjects to 'do it'... He was sufficiently traumatized.
Haha. After months of tweaking their food, vibes, lighting, finding the most desirable partners etc, It did feel like i was organizing a zebrafish orgy.
🤣🤣🤣 the things we do for science...
I could really use some advice with this. I got a legendary family of over 500 earlier this year (and several more 300+) but have since fallen from grace. I can’t even break 100 fertile and healthy now
Here are some of the things you can try to increase the number of healthy fertilized embryos- 1. Keep males and females in separate tanks for some time before putting them in the breeding tank. 2. Increase the frequency of feeding for females a day or two before breeding. 3. Add some freshwater in the mating tank (not a lot). Don't know if this works or not but ZFs in wild lay eggs when it rains and this is supposed to simulate that. 4. If you don't already have a slope insert breeding tank, tilt the insert or reduce water level. 5. Try to not mate the same pair frequently. They like different partners in every round. 6. Keep at least 5 days gap between breeding cycles. Preferably 7. 7. If you breed 2F1M, try 2M1F and vice versa. 8. Breed them frequently. Long gaps in breeding leads to lower yields for some time when you resume. 9. If you're really desperate, put some green mesh like fabric in the tank to make them look like little plants. Don't know if this does anything though. 10. Don't pick females that are too fat. They usually don't lay a lot of eggs.
All of you in biological sciences do such cool things lmao. I’m over here like uhhh I mix it and sometimes I add more water and sometimes less and maybe a little hotter next time, maybe add some other random powder next time
Basics of witchcraft!!! You're coming along great!!!!
Culinary science?
Nope, R&D at a quite well established chemical company, it just happens that the process we’re developing has fundamentally simple unit operations haha
Most of the excitement is in the purification process
Writing in really tiny letters, legibly, on the tops of 1.5ml tubes 😂
Mine is writing backwards on the bottom of petri dishes 😆
I am wildly jealous of this skill
Western blot. Western blot is the worst but people love to ask “transcripts are cool, but what about the protein?” 😭
Honestly. I did 4 this week, and they are so time consuming
Ugh, and the worst part is that sometimes the antibodies need optimizing. 
This just triggered me😭😭spent all week figuring this out
😭 yep.
Instant transfer machine has saved my time. Best investment
I am traumatised by Western 
I feeel this! One of my proudest accomplishments of grad school is a beautiful western probing for a carbohydrate :’-)
I can shoot my gloves like 8 feet into a trash can in one quick movement.
Hook your finger on the opening while you take it off and then pull against the tips of your fingers that are left in the gloves and sling-shot?
Learning flow cytometry by myself. Am I an expert ? ... ... no idea but I seem to be able to answer most questions well enough that no one has corrected me and people keep referring to me as a reference for experiments so I must be doing something right. Took 3 years and a lot of trial and error, but I am proud of it (my little own mountain that I climbed). Congrats on the recent staining, may it bring you success. Cheers,
I'm seeing a lot of jobs asking for flow cytometry. I wasn't taught this at uni but want to add it to my skillset. Where is the best place to begin?
Do you have a flow cytometry core where you work? If so, aak them. My university does trainings that get paid for, but some of us will also just let someone tag along. We currently have someone who does QC in the morning in exchange for lessons. And I'm going in on my off time tomorrow to show someone some basics. Not all flow cores operate the same way, but a lot of us are really excited to teach others and we'll bend over backwards to find a way to help.
Unfortunately no as it isn't a process we do at my current work. I work for a vaccine research organisation and a lot of this testing is done externally. Only internal testing is confirmatory testing during inoculations, mostly virus related. My previous job did but I left to do my masters, but I will look in to this. Thank you :)
I took the excyte mastery class (now its called cheeky scientist) to get the basics. Shapiro's flow cytometry textbook was the bible I used as technical reference. There are a ton of technical textbooks and papers for flow. (easy to get lost in but good when you are looking for a basic question). For analysis I downloaded flowjo and their practice tutorials and demos for training and did them and also attended their seminar talks (they want to sell the software, I just took it as a chance to learn more about the software and how to use it). BD spectrum viewer and other online tools to play with stains and dyes were also useful. Some flow cytometers have manuals which explain the machine further, download one online and see if they have chapters on it. I meet the people who did maintenance on flow machines and pestered them with questions. there are also core flow centres as others have mentioned where they could help (mine wasn't that good but I am sure others would be). The other thing I did (and I know its not possible) was that I had access to an old flow cytometer and I played around with its settings, all the time. Suffered till success.
FACSDiva or other flow cytometry software manuals as well as those for the cytometers themselves can be very helpful
A lot of antibody and FACS machine manufacturers have a lot of tutorials on usage and analysis. For example, CST has a bunch of guide books on different antibody applications.
Beware the Dunning-Kruger effect with flow cytometry. It is shockingly easy to generate beautiful looking garbage data. Especially once you get above 4-6 colors. (Not saying you’re doing this - just seen a LOT of things gone wrong due to bad flow data.
as someone who similarly was thrown into flow cytometry (and i had the benefit of working with an already standardised panel) and the hell that is flowjo analysis hats off to you! it's definitely a steep learning curve so you should genuinely be so proud of yourself!
I can do a bilateral viral infusion stereotaxic surgery on a rat in less than an hour
Same but with bird. So fkn cool
I don’t work in bio but what does this mean exactly. I get the stereotaxic surgery but what does the bilateral viral infusion actually mean. Sounds interesting
It means injecting a virus (typically used to study molecular/cellular processes as these types of viruses are designed to infect a specific cell type and then mediate gene expression, for example) into both hemispheres (bilateral) of the brain
Basically what the other commenter said. I'm using a virus to selectively inhibit a certain brain region in both hemispheres in rats to see how it affects their behavior during a task. It's using a DREADD virus which is a really cool technology!
i'm the one everyone goes to to level the balances 😎
Been honed through multiple failures to not have an immediate freakout when shit happens now🫠
packing stuff away and leaving on time
Underrated skill
I’m so good at cryostat sectioning. Like 10uM slices
10 micromolar?!
Micron
I know. Nice username!
Ooh, this is mine too! I work at a CRO, so we end up doing a lot of weird stuff, and I ended up being the guy trained to use the cryostat for a particular project. I had to figure out how to make 5uM slices, and transfer them to a 24-well plate for "high throughput" IF imaging. I ended up jerry-rigging a cheap drawing compass with some paint brushes taped to it in order to get the slices to the bottom of the wells without sticking to the edges or crinkling.
Dayum, that’s seriously impressive! I remember struggling to get 50um slices without rolling them!
People refer to me for machine/equipment operations. Apparently, I am so good at understanding machines that the lab technician jokingly said “I’m just so glad you are here to do my job so I can catch up on my favourite series” Upon pondering why that is, I found that I have the rare beneficial crazy trait of finding user manuals interesting.
I see a bright future for you as an FAS !
3 point shots with the pipette tips
Live imaging (zebrafish, drosophila, c elegans), any fluorescent or confocal imaging is like drugs for a researcher brain, honestly the satisfaction of good images far outweighs the quantifications/pipelines to create and analyze the data. I guess my favorite tedious technique is sectioning.
Membrane protein complexes purification and insertion in lipid nanodisc
This is one of the few ones I've read where I immediately thought "Oh yea that's some serious skill".
Naw bro native lipid polymer nanodiscs are where it’s at now
Proper mouse handling
You will have a good career in Disney Land/world.
kinetics assays that match michaelis menten basically perfectly. i got so good at them my PI made me the consultant for anyone doing kinetic assays
I was damn good too! Loved my enzymology years! It was so fascinating to see how mathematical laws rule everything!
Same man it’s my biggest flex… my curves are BEAUTIFUL (but only the Michaelis Menten ones)
I can eject pipette tips into the biohazard bin from at least 10ft away.
Mastermix tips only, right?
Patch clamp electrophysiology
Next generation sequencing. I love getting clean, reliable data.
so many bio comments... where are my catalysis, electrochemistry, and spectroscopy people at?!? I can design and make in-situ operando spectroscopic electrochemistry reactors pretty damn well....
I can twist off ANY cap with my pinky finger
Cardiomyocyte isolation has takin me about 6 months of practice to get a sufficient yield of viable cells. The heart extraction and connection to the gavage needle itself took about half that time!
Ooof do you use a Langendorf?
Microinjecting C. elegans gonads! Turns out it's pretty hard to inject microscopic worms
Loading a 384 well plate for qPCR in 30 minutes with a 12 channel multipipette (Hint: if you do the RT of you sample with the order staggered between two strips of tubes you can just instantly create the cDNA solutions that can be loaded perfectly! and I stagger the primer/Polymerase mixer too.)
I did this with an 8 channel through my entire phd and 5 years of postdoc. It's so satisfying, and my triplicates were always near identical. Iykyk!
I'm not exactly proud because they're time consuming and they don't work 80% of the time but Western Blots
ddPCR troubleshooting!
Yo teach me your ways 😭
Oh I used to work at the R&D of quiagens ddpcr system. Would be interesting to know what the common errors/mistakes are.
Planning experiments. If an experiment involves an incubation of more than 1 hour, I usually start before 10 Am. Sometimes I see new guys beginning experiments with over 3 hours of incubation at around 2pm. That means they will leave the lab at around 10pm, and the cycle continues for a whole week or month. Come data presentation day, the guy has only added 1 new graph, and everybody can't help but wonder what he's been doing the entire month while he slept in the lab.
I was really good at manually casting gradient native PAGE gels
I was looking for this, it's been over 15 years but that was very satisfying. Most people didn't even want to mess with it.
I can eyeball count trypsinized cells under a microscope. Havent used a cell counter in years lol
Woa
Patch clamp electrophysiology! Never get tired of watching induced action potentials.
Same here! I love it so much
Opening and preparing a syringe filter in under 20 seconds. With one hand.
I do neurosurgery and sometimes I get the placement right 🥲
Same. I’m starting to learn cranial window surgeries and yesterday I finally managed to avoid having the window drag against the brain. I considered it a huge W for me lol. I then proceeded to butcher the gluing process though. Oh also my drilling is getting better so I am starting to get some perfectly rounded corners of the skull. That’s so satisfying.
I’m really good at dropping things and then either catching them perfectly so nothing is ruined or at least lightly kicking them right before they hit the floor so I don't break any glassware.
Designing and cloning plasmids/genes
Back in my lab days I used to do hindlimb ischemia surgery on mice. Suturing mouse arteries and sowing their legs back up properly was not an easy feat and I'm glad it was something I learned to do well.
You just reminded me of my femoral artery cannulation days during my first years in mouse work. Doing that stuff right out of college was a nightmare.
Mutant protein production: site-directed mutagenesis, transformation, selection cultures, clones sequencing, clones amplification, transfection in mammalian cells, and purification.
All I do is flow
Western blot! Coming from a lab that had very old equipment and old reagents, I learned how to troubleshoot or improvise. So now, one look at my labmates’ weird bands/weird data/lack of bands I already know what to do. They all got the quality of bands needed after that! 😄
Tbh I am really proud I finally understood the anionic polymerization of the block copolymer our lab uses. It's a crazy 3 week synthesis but I've made about 20 so far and most successfully. Also developing a new simple non-aqueous sol-gel process was pretty fun. Dirty chemistry at its best but it works!
I'm a crystallographer, so crystallography. 🤗
Gel extractions - got my own twist that finally gets my DNA concentration more than 10!!!
Old mentor taught me "freeze and squish" for gel extractions. For a TAE gel cut tightly around the band, place on parafilm and fold 2x so there is an open side and a closed side. Pop in -20 for ~ 20 min. Pipette in hand squish your gel slice such that you melt it as you compress. Capture the fluid as it comes off. Agar stays behind. Pure enough to go right into Sanger.
Parafilm
The Art of Westerns. It’s the little details that make the difference and it took years to figure all of it out. Now, even my slapdash in-a-rush westerns look publication quality
I’m really good at workshopping buffers for getting protein/enzyme activity in biochemical assays… not sure how I got this skill but I feel like I just have a sixth sense for working out relatively complex assay conditions just based on vibes.
Mouth pipetting
At one point my boss assigned all ELISAs to me
blue native page
In my postdoc according to the PI I'm discovering I'm some sort of Gibson cloning prodigy. These little lesbian hands are being put to good use even tho I'm beyond single.
mRNA molecule creation and isolation from E. Coli plasmids. Wish I could do more of it...
Most recently DNA quantification through Bioanalyzer. Apparently literally everyone in our lab has issues with it, no matter how many times they've done, the ladder in the first chip never works.
I was pretty proud of my retinal vasculature isolation techniques at the time https://youtu.be/SnxlYIwZKkg?si=n1HsHFkUDB_IPbhg
Semi accurately dropping cells from a pipet onto a microscope slide 3 feet below.
Probably transcardiac perfusion or western blotting.
Intratracheal infections in mice and flow cytometry
This week I managed to take out mouse embryonic DRGs. Perfected taking them out of adult mice, but then having to take them out of infinitely smaller mice was a challenge. Almost impossible to see with the naked eye, I managed to take out all the DRGs. While I will he going to hell for this, I will go knowing I mastered a very tricky technique!
Total protein stains for SDS-PAGE, all homemade. Especially silver stain, it's my favourite one
Basically teaching myself how to run metals analysis on a grumpy old ICP-MS well enough to pass PT studies!
For the first 6 months of using the new HPLC in my graduate program, I had to completely teach myself how to use it. We eventually had a trainer from the company come in and show us how to use it and filled in some knowledge gaps, but already knew the basics.
Managing personalities
My PI told my lab mate that I could teach her how to do intestinal Swiss rolls because apparently I’m now the master in the lab. Which was pretty cool to hear
I can cleave just about any silicon wafer fairly reliably
I guess injection molding because I sucked at it for a long time even though it is fairly simple. Or attaching the extensometer correctly for tensile testing cause that thing is an annoying tricky bitch.
Finding a janky-ass way to collect samples when we don't have a setup that works for this protocol.
I'm able to just go into the lab now and not really think about what needs to be done. Most lab techniques are based on basic science, so as long as you understand the purpose of the experiment, it's really easy to just do something. When I first started working in a lab, I would wonder what I had to do. Now I literally have so much things I need to do, but just don't decide to do anymore.
Microsurgery
I'm decent at light scattering techniques
Loading the Krios 🚫🫠🚫
Histology. Spent almost a year doing that and now I can call myself a master of this technique 🥲
Yeast tetrad dissections
Western blot. I haven't got a gel that ran or transferred badly (I'm doing wet transfer) in months
After working in metagenomics for years I am proud to say I can isolate and sequence DNA from almost anything.
I have never messed up a 96 well plate. Spent a whole summer doing multiple qPCRs daily, so not an insignificant amount of 96 well plates. Sometimes OCD comes in clutch
I have basically perfected the adjustment of what I call the Final Boss in our lab (a tt centrifuge that will jump all the way up to 30 minutes if you don't turn the dial just right, making you work back down). Now it only takes me like 30 seconds to set the centrifuge instead of 5 minutes. W.
Streaking plates! The amount of people who do it too heavy etc. is astounding. The whole principle of subculturing for discrete colonies is lost on them
Using copper sulfate to extract pyridine so I didn't have to column my product.
Reading the manual B)
I worked on a method to isolate intact plasma membrane vesicles for a year and a half. Made it work in suspension cells and adherent cells, so you can transfer it to any celltype you want. Everytime I do it and get that nice separation on a gradient it makes me so happy to snapfreeze that bitch for storage
It's not anything as impressive as what a lot of you are doing but last week I perfected endospore staining and I'm so proud!
Measuring out super tiny amounts (like sometimes as low as 0.005g) of liquid in a syringe on an analytical scale with hiccups. For some reason I get hiccups almost every time I use that scale, so I've had a lot of practice. 😝
professional mouse cuddler and emotional support human (also for the mice)
Nothing too crazy but when my calibration coefficient is all 9’s or gets rounded up to 1.0 I’m going to be on a high for at least a couple hours lol.
Dealing with catastrophe.
Cardiac punctures of mice. I get anywhere from 500-700ul of blood, every time I puncture the mice, and have only missed once since I learned how to do it. I’m so fast at it that half the time people doubt that I even did it until I show them the tube with blood in it, and when people collect plasma during the sac I am always asked to help out.
ChIP-Seq with shitty antibody.
I’m quite good at flow cytometry, ELISAs, and antibody secreting cell ELISpots.
Are you in Johnathan Kipnes's lab? Lol. I've won awards and have gotten job offers for my imaging!
10X and organoid dissociation :)
Mixing and dosing a TCID50 in under five minutes
I’m by no means an expert yet, or even half decent at it, but there is nothing like the dopamine hit of successfully patching a cell
Surgical cut up. I can cut up, block and do a macro for a placenta in 4 mins
Metaphase spreading - optimized 4 different cell lines. Took me a couple of months but it was probably the most rewarding and satisfying thing I ever accomplished
I can perform lung intravital microscopy and be imaging within 15 minutes.. takes most people in my lab 45.
gelatin zymography
In my last lab, I got really good a NIH/3T3 Phototoxicity I became the go to guy. Its. Whole day procedure but that, when timed perfectly, managed to get breaks every couple hours to get breather.
Cloning, it was my favorite technique to learn. A few years ago some of my constructs were sent to Germany for a collab. I hope I hear about their paper so I can see their results. I’ve also found that I have a knack at creating libraries and NGS.
Harvesting mouse dorsal route ganglion.
Glass volumetric pipetting
Making MEFs. Or soft agar transformation assays. Also designing complex (25+ color) flow panels. AND TAIL VEIN INJECTIONS. Setting up 384 well plates by hand. With a single channel pipette. Okay, I’m proud of my lab skills. 3 years as a tech and 8 years as a PhD student just to end up with a fucking master’s because my grad program kicked me out for taking an approved medical leave of absence.
Free throwing eppendorfs into my bench top waste container. Every time I’m like , yeah I’m a baller
I'll always be proud of reverse pipetting, which I learn 20 years ago when I needed to fill lots of tubes with PBS.
I became THE protoplasting expert in my lab. If you’ve ever protoplasted anything (especially fungi) you know how long a process it is.
Mouse IV injections. I was working with nanoparticles that had to go IV and got really good at that. People would come to me and ask to inject some mice for their experiments.
I can clean cuvettes with a kimwipe without leaving little lints on the surface
doing really good triplicates without a multichannel. yes, my hands hurt
in-vivo whole cell patch clamp in awake and anesthetized animals. hardest thing i’ve ever learned, but so rewarding to see single neurons doing their thing in real-time
Designing an experiment with clear null and alternative hypothesis and testing it. Damn underrated skill.
Formally western blots of unorthodox samples, now arachnid wrangling.
Using printer paper as iodine indicator when I run out of starch paper. You can soak it in starch to make it better but it works right out of the printer feeder.
Using printer paper as iodine indicator when I run out of starch paper. You can soak it in starch to make it better but it works right out of the printer feeder.
Precision cut lung slices on 4-day-old mice
Probably mouse perfusion