Doug Shepherd

@dpshepherd.bsky.social

recovering alpinist and scientist.

If you've seen anyone from the lab talk about RNA-FISH over the past years, they've shown data somehow related to this preprint. After a wandering road writing it up, happy to put out some thoughts on scalable 3D imaging-based spatial transcriptomics data processing. www.biorxiv.org/content/10.1...

GPU-accelerated, self-optimizing processing for 3D multiplexed iterative RNA-FISH experiments

Imaging-based spatial transcriptomic approaches rely on iterative labeling and imaging of carefully prepared samples, followed by solving a computational inverse problem to determine the location and ...

biorxiv.org

We are really enjoying having Nick Galati visit us on sabbatical for some fun protist work, including our first forays into expansion microscopy! Max projection xy&xz views of expanded paramecium and tetrahymena cells. Scale bar is ~5.5 um accounting for expansion. Epifluorescence + deconvolution.

Pseudo-colored fluorescence microscopy image of expanded paramecium and tetrahymena cells.

Our special issue of Phil Trans Roy Soc A on "Biological Fluid Dynamics" Emerging Directions" is officially out now! Originating from discussions at one of our SIG meetings, this special issue has 16 research articles and reviews on all things bioactive! royalsocietypublishing.org/toc/rsta/202...

Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences: Vol 383, No 2304

royalsocietypublishing.org

Steven has our Snouty-OPM running again after our lab move. He added projection mode, sensorless AO, and a new control GUI based on @talley.codes + crew's pymmcore-plus/pymmcore-gui. Here are some IF-labeled microglia (orange) in the middle of an uncleared 60um mouse brain slice. No deconvolution!

Immunofluorescence image of a mouse brain slice, with nuclei (magenta) and microglia (orange) shown.Zoomed in Immunofluorescence image from the full mouse brain slice, with nuclei (magenta) and microglia (orange) shown.

Out of nowhere, our institute just froze all indirect accounts - no matter when or where the indirects were generated. With completely unclear direction on how this works going forward. No words…

In a brief respite from current events, I got to see a California Condor for the first time while running in the Grand Canyon this weekend! What an amazing animal. Unfortunately, public lands and those who protect them are also targets of this admin. Keeping fighting…

California condor perched on a rock with the Grand Canyon in the background.

Anyone successfully dealt with chronic, at times crippling, facilities issues? It has really impacted the team's productivity, even though it's not their fault. I'm open to any and all advice beyond moving spaces. We already did that twice in 5 years with new (broken) promises each time.

We haven't imaged big samples in a few years. Recently Steven brought our custom ASLM, with some fun/interesting detection optics, back to life. Tatz Murakami graciously sent us a beautifully cleared and vasculature labeled mouse brain for imaging. Fused max projections of raw data.

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Today's adventure in academia. How many layers of IT do I need to contact to reset the fancy new Zoom TV system they put in our conference room. I'm betting greater than ten, but there are a still a few hours left in the day!

Fluorescence Friday! 3D timelapse of MOLM-14 cells expressing GFP-LC3-RFP embedded in collagen. Orthoview rendering of GFP (cyan) and RFP (orange) imaged every 30 seconds using our Snouty-OPM.