🤔 #AskZebrafish: "Do any of you have a protocol or a kit recommendation to do subcellular fractionation of zebrafish embryos. Ideally I'd like to separate the mitochondria and the nucleus (and as a bonus the rest of the cell parts)"

🤔 #AskZebrafish: “Wondering if anyone has recommendations on a relatively inexpensive device to measure water flow rates in a swim tunnel?”

#AskZebrafish from Slack: What is the best promoter to use to specifically tag microglia in #zebrafish? Thinking of using mpeg1.1 but just wanted to double check if there are any better options?

Query from Slack: Looking for Tg(pax3:GAL4) and any driver line for ascl1:GAL4 or Cre? Preferably in Europe. Any help and/or leads would be appreciated! #AskZebrafish

🤔 #AskZebrafish: Looking for a zebrafish version of the Mito-RiboTag mouse line. Has anyone made one or might have a vector they could share? Any leads would be thoroughly appreciated!

Question from Slack: Looking for antibodies against mscarlet-i that people have good experience with? #AskZebrafish

Query from Slack: Looking for notch2 mutant zebrafish and someone willing to share them? Preferably in the US. 🔍 #AskZebrafish

#AskZebrafish query from Slack: Looking for contact that has recently shipped transgenic eggs from Europe to the US. Re: any hurdles you or your lab had to go through. If this describes you or someone you know, please reach out 🙏

Initial trawl came up empty 🤷‍♂️ Any leads or feedback appreciated!

Query from Slack: “I was wondering if anyone has a spi1b (pu.1) HCR probe that looks good in whole mount [zebrafish] tissues, the one I designed has a lot of pesky background in the embryo.” #AskZebrafish

Query from Slack: Has anyone successfully used a c-fos antibody for whole mount immuostaining in zebrafish? #AskZebrafish 🤔

Two queries from Slack: Anyone know of any good antibodies that work well in whole mount 5-7dpf for labeling mitochondria? Does anyone in the community work with mitochondria based assays for larval zebrafish? Such as Alamar blue or JC10? Or anything else? Any leads appreciated! #AskZebrafish

🤔 #AskZebrafish from Slack: "Does anyone have a favorite protocol for staining myelin in the brain, especially in fish >21 dpf? We would like a dye, not a line. We're looking into CMC or fluoromyelin but are unsure about penetration into the brain." Any feedback or leads would be appreciated!

🤔 #AskZebrafish

#AskZebrafish from Slack: For folks studying metabolism, have you done chemical screens in recent years? If so, which libraries will you recommend? 🤔

#AskZebrafish: Does anyone know of a good antibody recognizing mTagBFP2? Preferably available in North America. Any help would be appreciated 🙏

Query from Slack: Looking for any transgenic lines that have integrated and expressed Cas9. Preferably in the US. 🤔 #AskZebrafish

Query from Slack: Looking for a line that labels the nuclei of basal epithelial (and migratory) cells. Preferably in Europe. The best line seems to be one the Sagasti Lab made: p63[BAC]:Gal4 crossed to a line for expression in the nucleus (e.g., UAS:nls-GFP). Would consider others. 🙏 #AskZebrafish

Query from Slack: Does anyone know of good commercial centromere and/or kinetochore antibodies that work well in zebrafish? #AskZebrafish

Brainbuster from Slack: Has anyone done agilent seahorse mito stress test on #zebrafish embryos (2-3dpf) and be willing to share a protocol? #AskZebrafish

Query from Slack: Does anyone have an anti-S100b antibody that they are successfully using for IHC in whole-mount larval brains. The one most commonly used one (Z0311 by Dako) seems to have been discontinued. Any pointers to a good one to use in fish would very helpful! #AskZebrafish

Query from Slack: Looking for someone to share somitabun mutants in Europe? Any leads would be appreciated! #AskZebrafish

Query from Slack: Looking for people with experience in proximity labelling in zebrafish? Considering using miniTurbo instead of BioID or APEX, what do the community think of it? Also looking for anybody that has experience with targeting labeling enzymes to the ER/secretory pathway? #AskZebrafish

🤔 #AskZebrafish: Would anyone within the EU have a mutant ybx1 line swimming around and willing to share? Any leads would be appreciated.

🤔 #AskZebrafish: Looking for krt18a reporter fish in the UK or if anyone has a ready to inject construct for making one and are willing to share, that would be much appreciated!

🤔 #AskZebrafish: Looking for Prom1a and/or Prom1b mutant lines and/or reporter lines that someone would be willing to share? Preferably in the USA.

🤔 Query from Slack: Looking for 5' myl7/cmlc2 promoter vector and someone willing to share it? Preferably in the US 🐟🫀 #AskZebrafish

Another query from Slack: Looking for ccdc40 mutant #zebrafish (aka locke). Preferably in the US. Any leads would be much appreciated! #AskZebrafish

Query from Slack: Does anyone have good recommendations for antibodies that label mitochondrial DNA encoded proteins and work well in #zebrafish? #AskZebrafish

Anonymous query from Slack: Does anyone have the TgBAC(foxp3a:TagCFP-NTR)vcc5 line swimming around, preferably in the US? Thanks in advance! #AskZebrafish 🤔

Still on the hunt for fish lines 🙏

Still searching for leads! 🙏

Head-scratcher from Slack: Does anyone know if it is possible to tell (by eye/microscope) if the developing oocytes in a zebrafish ovary are at the pre-, during, or post-vitellogenesis stage? 🤔 #AskZebrafish

Looking for anyone w/ experience in smRNA analysis who can provide someone with information or guidance? They already have their sequences, but can't find a clear methodology/protocol to help them analyze it. The sequences were obtained from NOVOGENE's Small RNA Sequencing service. 🤔 #AskZebrafish

Any leads would be appreciated 🙏

Any leads would be appreciated! Reposts welcome 🙏

Still searching for leads 👀

Still looking for leads 🕵️

Query from our Slack Tankspace: Does anyone in Europe have the Tg(mpx: mcherry ) line from the Huttenlocher Lab or similar and would be willing to share? #AskZebrafish

Still looking for leads 🕵️

Re-upping

Another query from Slack: Does anyone (preferably in the US) have the Tg(GFAP:iGluSnFR) line and be willing to send them to another #zebrafish lab? 🤔 #AskZebrafish

🤔 #AskZebrafish

Still looking for guide sequences! Any leads and/or help appreciated!

🤔 #AskZebrafish

Query from Slack: Does anyone have scn8aa and scn8ab #zebrafish mutants and would be willing to share some embryos? (Preferably in Europe, if possible) 🔎 #AskZebrafish

🤔 #AskZebrafish

Re-upping 🙏

🤔 #AskZebrafish

Re-upping