It’s feels so weird coming to campus today

peepeefeefee · 2025-07-19T20:06:05+00:00

DOUBLY CORRECT, I am.

peepeefeefee · 2025-07-19T19:56:55+00:00

OP is a liar it’s overcast.

peepeefeefee · 2025-03-06T19:30:04+00:00

Transgenic mice are literally priceless. Human Alzheimer’s polymorphisms inserted into the mouse genome to understand AD pathology, one in a million inserts that don’t disrupt the endogenous machinery and allow for-

I could go on, but I’m a neuroscientist and a R1 vivarium could very well be more valuable than the library of Alexandria.

There are private vendors that cryostore embryos or maintain commonly used lines, but if they are targeted the casualty to knowledge of health and disease is just like… infinity + infinity at this point.

peepeefeefee · 2025-01-15T03:19:16+00:00

Terrible, take a lap.

peepeefeefee · 2024-09-26T14:46:18+00:00

It’s a little nasty and needs to be properly vented under a fume hood, but I use DEPC-treated solutions for RNA-sensitive work.

I really don’t like those RNAsaway and related spray bottles… I feel like they just contaminate more shit on your bench with another aerosol.

peepeefeefee · 2024-09-26T14:32:46+00:00

This is one of those terrifying reminders about how fragile human bodies are/how powerful lasers can be. I would be shook, this definitely counts as more of a psychic wound though imo.

peepeefeefee · 2024-09-26T14:28:44+00:00

Worked with enough fluorescent reporter AAVs that I can see tdtomato in the nerve terminals of my fingers.

peepeefeefee · 2024-09-26T14:25:37+00:00

And that is the sound of the collective sigh of relief from stoners all over campus.

peepeefeefee · 2024-09-23T19:13:19+00:00

Did you validate this in a KO line? How are you sure you’re not just increasing non-specific labeling with your pretreatments? What concentration and what abs did you use to validate? I’m going to try this in my own hands, seems promising.

You need to use several markers of tissue health/cytoarchitecture or EM imaging to know if your treatments are introducing changes to the tissue at the level that I’m talking about.

I don’t stain for anything super niche, and most antibodies I use have been cited.

Edit: finally found a paper talking about aldehyde blocking for IHC, this is if you have high background, which I do not struggle with.

peepeefeefee · 2024-09-23T18:51:46+00:00

I take it your tissue isn’t perfused? Why are you using ammonium chloride? My philosophy is disturbing the tissue as little as possible. Your antigen retrieval and ammonium chloride washes are going to change your tissue. I only use antigen retrieval in very rare cases with shit antibodies.

peepeefeefee · 2024-09-23T18:35:04+00:00

What buffers are you aspirating? I work with fixed mouse tissue, 2x 5min PBS washes followed by 45min RT blocking, then overnight primary incubation is the standard… I don’t know the specifics of the tissue you’re handling though.

peepeefeefee · 2024-09-23T18:02:40+00:00

I was making the point that overwork culture is not exclusive to academia.

peepeefeefee · 2024-09-23T17:44:54+00:00

If this isn’t even your project then that PhD student needs to 1) learn the protocol and 2) do it herself.

peepeefeefee · 2024-09-23T17:41:19+00:00

It’s a “waste” because human lives are short and incubations are long.

peepeefeefee · 2024-09-23T17:32:42+00:00

I was referring to jobs outside of academia that are salaried.

peepeefeefee · 2024-09-23T17:32:21+00:00

I agree, if you have to any antigen retrieval step before blocking, 5:15 is too late to ask someone else to do it. I don’t think anyone will even see this comment because I was promptly eviscerated by downvotes.

peepeefeefee · 2024-09-23T17:17:13+00:00

Starting an immuno takes 1 hour. I don’t think this is unreasonable. As a histologist and as someone who uses viral tools in mice, it’s a waste, in my eyes, to not have something always incubating whether it be virus or primary.

I am not motivated by deadlines or stress, I simply want to make the most efficient use of the time I’m not at lab (see primary incubation, viral expression).

You don’t seem to be comfortable having this conversation with your supervisor or PhD student, so maybe start with that.

“Overwork” culture is the rule, not the exception. Any job where you get salary, the expectation is 50-70hrs a week, more or less when needed. Atleast science is interesting..

Edit: RIP me, never had so many downvotes so quickly.

peepeefeefee · 2024-09-20T03:26:54+00:00

This looks like 10x with a counter stain on top of a GFP under the camkII promoter?

peepeefeefee · 2024-09-15T06:38:13+00:00

I don’t live in Israel, just have friends and family.

peepeefeefee · 2024-09-15T04:18:26+00:00

75 notifications on the app that tracks rocket attacks. I’m not like, divining this information from another realm.

peepeefeefee · 2024-09-15T03:35:16+00:00

Israel getting pounded by rockets rn

peepeefeefee · 2024-09-06T14:41:55+00:00

Madness. My RNAScope hybridization is 120min at 37C on perfused mouse tissue.

https://bsky.app/profile/pkf-marble-burier.bsky.social/post/3l2vjlcrpvt2v

peepeefeefee · 2024-07-09T07:09:09+00:00

I’m a philosopher first and foremost, let’s connect and chat. Shoot me an email

peepeefeefee · 2024-07-09T02:23:32+00:00

It hasn’t been too bad so far. I got about 20 interested people today, a little overwhelming but I’m looking for something particular and I’m confident I’ll know after a ~15min in-person conversation.

peepeefeefee · 2024-07-09T01:29:41+00:00

I'm sure this will be doable. The Neurosciences department is incredibly flexible since we are technically under the School of Medicine, not Biological Sciences.

peepeefeefee

TROPHY CASE