What if you got to work with some of your best friends on a science project? I can't publish how fun this was, but I can show you the data (🧵)! Last week, we posted our second neutrophil swarming paper to bioRxiv and I wanted to post my favorite videos here~
www.biorxiv.org/content/10.6...
Snouty objectives enable high resolution light-sheet microscopy with standard microscope platforms and sample formats!
-> Here are citable repositories with CAD, Zemax and summarized optomechanical specifications:
AMS_AGY_v3: a.k.a. Empress Snout, now the best!
github.com/amsikking/AM...
👍
@patrickzager.bsky.social acquired this lovely 3D timelapse of live neutrophil membrane protrusions on the snoutscope at the UCSF Center for Advanced Light Microscopy. Shown are three orthogonal maximum intensity projections.
Hi microscopy folks! I'm looking for your advice 💡
We're shopping for cyclic immunofluorescence instruments to increase throughput and multiplex-ability of tissue IF. Have you worked with cyclic IF platforms (Comet, CellScape, Cell DIVE)? Thoughts? Pain points? Alternatives to consider? Thanks!
The power of CAD! The ability to precisely draw 3D objects and have them made is still a miracle to me!
-> Here's an old design for an alloy block to hold microcentrifuge tubes near freezing when moving them around. It's simple, but I wouldn't want to draw it or make it by hand...
Celebrating science with art! Every 2 weeks for 1 year, the full alphabet. Like, reply or repost with your favorite "Z" scientist!
(26/26) -> "Z" is for Zebra, Zucchini, and...Zernike?
en.wikipedia.org/wiki/Frits_Z...
Art by @annesikking.bsky.social (my Mum).
1: with no tilt on Snouty the beads will come in and out of focus as you expect. 2: with a 90deg tilt you'll traverse the beads along the coverslip (low transmission but conceptually useful). 3: With a typical tilt of 30-45deg you'll get some of 1 and 2 (which looks like beads moving vertically).
Proof of principle. A critical step in many projects...
-> Here's the first data from an AMS-AGY objective. Without building the whole instrument, it showed that a tilted Snouty (Frustum) objective can avoid the resolution lost in an oblique plane microscope!
amsikking.github.io/high_na_sing...
Celebrating science with art! Every 2 weeks for 1 year, the full alphabet. Like, reply or repost with your favorite "Y" scientist!
(25/26) -> "Y" is for Yak, Yam, and...Young?
en.wikipedia.org/wiki/Thomas_...
Art by @annesikking.bsky.social (my Mum).
Yes I wish the FOV on Nikon (and other) objectives was better specified. Some models are labelled OFN22, others OFN25. The new TIRF series is supposedly compatible with the new Ti2 platform (25mm FOV) but I can't find a definitive spec.
More thoughts here:
amsikking.github.io/microscope_o...
Our snouty v3 (AMS-AGY v3) objective arrived today! Looking forward to getting it installed on our single objective light sheet system very soon...
How has AI changed CAD? Can AI now reliably generate 3D parts, assemblies and drawings? I'm interested to hear about new tools, workflows and experiences ;)
-> Here's a series of enclosures I designed years ago for housing custom lab equipment. Can AI CAD just generate this automatically now?
Celebrating science with art! Every 2 weeks for 1 year, the full alphabet. Like, reply or repost with your favorite "X" scientist!
(24/26) -> "X" is for Xeme, Xigua, and...Xanthopoulos?
en.wikipedia.org/wiki/Basilis...
Art by @annesikking.bsky.social (my Mum).
I'd say it's as sterile with SALDA as it was without SALDA. The SALDA holding tube (stainless steel) and delivery tube (e.g. PTFE) can be sterilized. The delivery tube is readily replaced for an new one if contamination from a previous liquid is of concern.
Special thanks to co-authors @nybergg.bsky.social, @jlazzaridean.bsky.social and Elliot Mount for all their help and hard work!
This project was funded by Calico Life Sciences LLC and was made possible with the kind support of Magdalena Preciado López and Robert Keyser.
www.calicolabs.com
A SALDA fluidic system is readily cleaned or replaced and can dispense volumes as low as 10 µl with high trueness (RB = 0.32%) and precision (CV = 1.63%).
-> Here we show 100 random dispense events in the 10 µl to 100 µl range with cell culture media with < ~3% error.
A SALDA is cost effective and readily deployed on commercial or custom microscope platforms.
-> Here we provide a SALDA for a specific stage top incubator and inverted microscope, but the setup is easily modified and applied to any system that can accommodate a small ~2 mm diameter tube!
A SALDA enables accurate and repeatable timing of dispense events without user intervention.
-> Here's a SALDA on an inverted microscope with 488 nm excitation and a stage top incubator. The arrangement can readily access pre-programmed wells in a 96 well plate and dispense in < 1 s! (3x playback)
Adding a liquid to a sample during imaging and observing the response is challenging, and often done by a person awkwardly hovering over the microscope with a pipette.
-> Here we show the fluorescence response from a SALDA adding thapsigargin to U2-OS cells. User input and disturbances are avoided!
Sharing: A simple automatic liquid dispense arrangement (SALDA) for timelapse microscopy.
-> A SALDA enables accurate and repeatable liquid perturbations during timelapse microscopy and avoids environmental disturbance and vibrations from the user!
Details here:
amsikking.github.io/SALDA_for_ti...
Celebrating science with art! Every 2 weeks for 1 year, the full alphabet. Like, reply or repost with your favorite "W" scientist!
(23/26) -> "W" is for Worm, Watercress, and...Watt?
en.wikipedia.org/wiki/James_W...
Art by @annesikking.bsky.social (my Mum).
Done!
Pro tip! As much as I like the 55deg #Snouty microscope configuration (previous post) it's hard to align and keep aligned!
-> Consider using pre-designed O2-O3 optomechanics and an eyepiece to help avoid collisions during the set up!
github.com/amsikking/SO...
Celebrating science with art! Every 2 weeks for 1 year, the full alphabet. Like, reply or repost with your favorite "V" scientist!
(22/26) -> "V" is for Viper, Vanilla, and...Volta?
en.wikipedia.org/wiki/Alessan...
Art by @annesikking.bsky.social (my Mum).
I'm excited to be hiring an intern to develop new fluorescence-based assays for alpha synuclein aggregation at Calico during summer 2026 😃 🔬
If you're a current PhD student who loves microscopy, unnatural amino acids, and/or synuclein biophysics, please apply!
www.calicolabs.com/careers/?gh_...
AMS-AGY v3 objective replica
The ASI machine shop made a nice replica model of the AMS-AGY v3 objective for us to have in the #CellBio2025 booth. The first of the real ones will be ready by the end of the year.
Hands down my favorite #Snouty microscope configuration!
-> The air objective with a 55deg light-sheet. It's not the highest resolution version, but the convenience of the air objective, the large field of view and the steep sectioning of the light-sheet make it a pleasure to use!
#AIRR #miOPM
