u/Immediate_Editor9036

Hi, i was wondering if anyone here is familiar with mast cell biology. i have been doing immunophenotyping in muscle and lungs of balb/c mice. For some reason, the CD117+ Siglec F- cells that i have assumed are mast cells are not expressing Fcer1. I have done this with both control steady state mice and virus infected mice. However, my basophils are positive for the FceR1 receptor so i know the antibody works. i have seen this every time i've used this panel. i'm confused because im under the impression that mature mast cells in tissues should express FceR1.

last week i attempted having sex with my bf of about 1 year. i left highly conservative religion last year so i definitely needed time to get comfortable with the idea of having sex outside of marriage. we both had never had sex before. we have been taking things very slow until we both felt we wanted to try having sex and i was really excited about it. we had been intimate in other ways but this was our first time trying penetration.

we used lots of lube and i'd say i felt pretty relaxed, but when he entered me all we could fit was his tip and maybe slightly more. it was painful for me. its hard to even describe the pain. i first felt it at the opening of my vagina and then when he tried to go deeper i felt like he was pushing against something solid in my, it was so weird and that caused more pain.

i also tried inserting fingers inside me and i could do it with no issue. i could insert 2-3 fingers but then when he tried to insert his penis it just didn't work... we tried for a bit before we decided to stop. i know i should see a gyno now, but wait times to see one can be like 6 months. is there anything i can do in the mean time thats safe and effective? i really want to be able to have pain free sex. right now it feels impossible for me to have sex that actually feels good.

hi, I am just about to wrap up my first year of an immunology/virology masters degree. I need some advice on how to stay motivated. over the past month or so, I’ve noticed that I’m starting to have hard time with school. on days where I don’t have big experiments or early meetings/commitments, its hard for me to get out of bed in the mornings so I end up being late. I feel really bad about this because I know that with grad school you get back what you put into it. I’ve had lots of busy days so far and I enjoy being productive in the lab but now I just feel like I’ve fallen flat. I feel like I’m dreading going to the lab but I can’t even really figure out why. I have 2 supervisors who have been good and both my lab environments have overall been good too. I’m just trying to force myself out of this slump I’ve been in. I feel exhausted everyday and like I’m in this fog. I do have a few weeks off coming up in like mid August (encouraged by my supervisor since he recognized how busy I’ve been and that I’ve been struggling a bit). So I’m not sure if I just need a break and then I can start fresh in the fall. But I’m worried that this will continue even when I get back. any advice is appreciated!

honestly, I am just heartbroken. I was diagnosed with AGA last year and have been treating it ever since. although I’ve seen progress with my hair, a narrower part and lots of baby hairs according to my derm, I am so sad about the hair that I lost. I’m only 23. even though my derm says I’m in the very early stages, it feels like I’ve lost a lot. I have curly hair and my curls just aren’t the same. they used to be so long and full and easy to style. now I just don’t even know what to do with it anymore because since thinning my hair has changed so much so I just wear it up pretty much everyday. I am so angry and this just doesn’t feel fair. my hair will never be how it used to be. 1 year in and it still hurts. I am so jealous of girls who don’t need to think about this. I am so anxious about whether or not treatment will work for me long term and what my hair will look like years from now.

I feel like before leaving faith I would be so scared at even just the thought of not believing in god anymore. it was so terrifying once I started to deconstruct. they villainize the “world” so much and I fully believed that I could not find true joy or happiness without god. I was also scared that something bad was going to happen to me so that god would bring me back to him (if that makes sense). now, 10 months later, I see now that they say those things to control people and keep them in. I can happily say that literally nothing happened when I left, and if anything my life has gotten better. I’m in a loving relationship with someone who isnt Christian (this is something I would never have even considered a few years ago) and I feel even more connected to the people I love because I no longer need to believe that my love comes from god. it’s mine and mine alone. I’m even more in awe of this planet that we’re on and the billions of years it took for us to get here. there is so much more beauty in the world once you remove religion.

hi, I’m in my first year working with mice. i started in september. early on i got bit by a female B6 mouse when i was trying to practice restraining. I hadn’t scruffed her yet, I was holding her tail and she turned around and bit my finger. she bit hard and for a while so it broke through my glove and skin. after that I was really scared of working with mice. it’s gotten better as I’m now able to pick up mice and handle them normally without too much issue. but the problem is scruffing. so far for my experiments I’ve only had to work with sweet and nice balb/c mice. I’m comfortable restraining them since they literally never bite, but I’m still terrified of restraining B6 mice and I don’t know how to get over this. the good thing is I probably won’t be working with B6 for a little while still, but there are certain techniques that I can’t get trained on (like IP injections) until I’m confident with scruffing more aggressive mice. I feel like maybe my technique needs improvement but whenever I practice with balbc mice, I feel like my fingers always end up in a way that I know a B6 be able to bite me. or they wriggle away and I need to scruff multiple times to get it done properly. does anyone have any advice? I feel bad about not being able to do this yet because I think I’m at the point now where most others would be confident with scruffing …

hi, I have a big experiment tomorrow. 16 mice and multiple tissues and I will be doing peptide stimulation and then staining for high parameter flow. for the high parameter flow, it will be a panel with 30 antibodies. to save time tomorrow I’m wanting to prepare my antibodies today. that way I don’t feel rushed and there’s less chance I’ll make a mistake. my panel has tandem dyes and brilliant dyes, but I will be using a tandem stabilizer and BSB. will this be okay?

hi, i am 23 and i have aga. i have been on topical minoxidil and 150mg spiro since august 2025. I am still having issues with irregular menstrual cycles and spotting, as well as prolonged periods lasting a few weeks.

the last time i saw my dermatologist, she said if my cycles get to be too much for me then i can take birth control to regulate my cycle. i have never wanted to take birth control because i'm really worried about side effects, but i feel like my irregular menstrual cycle is just so frustrating for me that i'm debating taking it. my cycle used to be perfectly regular and predictable and it always lasted 5-6 days, and i miss that predictability. i'm wondering if any of you have ever had this experience and what BC has been like for you

i have a wedding coming up in a few weeks that i was invited to a while ago. since sending in my rsvp, a lot has changed. ive left the church and im dating a non christian. everyone has tried to reach out many times to get me to come back but im just not interested. i feel really nervous about going. my boyfriend will be my plus one so he will meet everyone there. i also had a really bad friendship breakup with someone who was literally driving me crazy with her self righteousness. she would text me constantly about wanting a biblical friendship or whatever and then would never ask to do anything with me and always cancel plans. there was a lot more that happened but now we are no longer friends because she ended the friendship with me. this happened a few months ago and im honestly still angry. she will be there at the wedding and i'm not looking forward to seeing her. Also, im not looking forward to the judgement and prying questions about my dating life or why i haven't been at church. this is definitely the last time i will ever set foot in that church and im just worried about the unpleasant emotions it's going to bring up.

It’s like 1am right now and I’m panicking about an experiment I have next week. I’m a 1st year masters student and this is my first really big experiment so I really want it to go well. this experiment was moved up so I feel like I am scrambling to prepare

my concern is how big this experiment has gotten. In short, it’s an animal experiment with 15 mice. at first I thought this would be easy peasy. I will be taking spleen, popliteal LN, and inguinal LN from each mouse. I will be performing peptide stimulation of all these samples with 2 different peptides. this all adds up to 90 samples. Then with all my controls for the peptide stim and for flow, I’ll probably be at about 100 samples. i thought that this would be manageable, but for my lymph nodes I can’t use 96 well plates because the pellet will be so tiny, especially because I will be splitting up the cells to stim with both peptides. So my supervisors told me to use eppendorf tubes instead.

I simply don’t understand how I can manage this. I’m used to long days in the lab, but this seems kind of crazy. Our micro centrifuge also obviously cant hold that many samples so every wash step I do will require multiple spins (probably like 5).

I need advice. is this kind of experiment normal and manageable? should I talk to my supervisors? should I switch to using 96 well plates? I would just be really worried about losing sample

I know not all churches are like this. but I went to a Pentecostal church in Canada for a long time (part of PAOC), and all the churches in this denomination as well as denominations that are similar, are SO showy and extravagant.

I’m questioning this more now that I’ve left. I served in many ministries, but worship was my main one. I played multiple instruments and I’d sing, I also would MD. the amount of time and money that was going into turning everything into basically a perfect concert was absolutely insane.

we all had fancy in ear monitors, we had a crazy $500,000 sound board, the tech staff EQ’d the entire auditorium, every service, song, prayer moment, sermon, was extremely timed. we had lots of expensive cameras for online streaming, and I was just watching a recent sermon just for fun and the pastor was saying that they are going to start expanding, no DOUBT by using tithe money. then they say ”the lord will provide“ whenever they talk about their financial debts, when really it’s the congregation that is going to provide.

the whole thing just bothers me. it’s just a way for people to get onto a stage, it has very little to do with god. and this is coming from someone who liked worship because I like playing and singing for an audience.

it makes me think, if god is really enough, why do you need all of this?

ok so I recently started therapy to help me work through the emotions and anxiety I have had since leaving the church. I’m 23 and started going to an evangelical, fundamentalist, Pentecostal church when I was 15. the therapy session was seriously so helpful and insightful for me and I felt really understood and I just wanted to talk about it.

I struggle a lot with the fear that I’m making the wrong choice or turning my back on god. I’m also in a relationship with a non Christian and this complicated things even more for me because I used to hear so many warnings about dating outside of faith, I can’t even describe the internal conflict and guilt I would feel when we first started dating. And not to mention the shame I felt when we first became intimate. I stopped going to church because I decided I didn’t want to feel guilt anymore, but then that caused the worry that I was avoiding accountability, because as the say, “iron sharpens iron”

My therapist really helped me understand that the guilt and shame that I feel about some of the decisions I’ve made isn’t even coming from my idea of “god”, it’s coming from the people at the church, and it’s their voices in my head that I hear whenever I feel like I’m doing something wrong, and it’s the fear that like I’m going to be rejected and ostracized by a community that sunk its claws into me and formed my beliefs when I was a teenager which is a really important time in life for creating your own worldview.

when I first left, I had a few friends tell me the best thing to do is end my relationship. I had another ex friend tell me that people were talking about me at church and her response was that I “never got to make mistakes” so I should be allowed to do this. this friend also would pray for me because ”the enemy is placing lies in my head”.

the more I thought about these things, the angrier I got. why do they get to tell me how I should live my life? whenever they reach out to me and ask to see me I can always sense that it won’t be just a normal hangout like it’ll be a push to try and get me to come back because in their minds, god is using them to speak to me and they are right and I’m wrong and I’m making a mistake. they think they are divinely anointed so they understand things better than I do and it infuriates me. They think that because they prayed and my name popped up in their mind, that must be god telling them that I’m lost and they need to save me. Like why would god talk to YOU about me but he would never talk to me when I would try to pray ?? Like leave me the fuck alone for real. They are so self righteous

I left in October. On sundays I still feel weird and guilty about not going. people and pastors from church check in with me out of concern or to meet with me. I just got invited to a grad party next week, my old youth pastor’s husband just got a doctoral degree. I felt really weird receiving this invitation, I don’t know why. I care about both of them and I think they are nice people but I’m not sure if I want to go. it would be hard to face everyone. ive only talked to 2 people from church about my change in faith. one of those people is no longer my friend, the other reacted kindly. for everyone else I just say I’m busy with my masters degree. Sometimes i feel like I’m making a mistake by choosing a life away from religion and church, even tho I know its for the best in the long run.

Hi, does anyone here have tips for perfusing a mouse lung through the heart? I've tried multiple times now and im never able to get the "white lung". My PI tells me it should be easy, but the one time he showed me he also wasn't able to do it and no one else in my lab is working with lung tissue.