Cell biologist interested in organelles and how they move. Husband, dad, intermediate filament apologist, and postdoc in the
@JLS_Lab
at
@HHMIJanelia
.
#Mitosis
. Here are a few angles of a dividing HeLa cell. Microtubules are white and the chromosomes are orange.
Imaging modality = Spinning disk + deconvolution.
In a recent overnight imaging session, I made a startling discovery... COS7 throw midnight dance parties. Who knew? These party animals are dressed in Vimentin-TagGFP (orange) and the ER marker SnapTag-Sec61b (blue)
#FluorescenceFriday
Mitosis montage. Orange = chromosomes 🧬, white = microtubules. Not all stages are represented, but left to right shows the general progression from prometaphase to late anaphase. These are max projections of HeLa cells (from deconvolved spinning disk stacks).
It's like a real-life game of snake 🐍. Check out this
#mitochondrion
chasing its own tail.
Mito label = TMRM.
Cell type = COS-7
Scale = 10 µm.
Time = mm:ss.
Bonus content! I don't think this movie made the final paper, but I still think it's one of my favorite examples of a metaphase mitochondrion pushed around by an actin comet tail.
cyan hot = mito
orange glow = actin (lifeact)
Loop ~40sec.
More here:
Thank you to
@NikonSmallWorld
and this years judges for selecting my neuronal actin movie as a winning video! I'm incredibly honored. Check it out and all the other winning videos in the link 👇
My lattice light-sheet movie of actin in a dividing cell (acquired at the
@AICjanelia
) was named an honorable mention in the 2019
@NikonSmallWorld
in motion contest! Hela + Lifact-EGFP. Rendered in
@ImarisSoftware
Overjoyed/honored/relieved to share our new paper describing multiple actin networks that position and shuffle mitochondria before cell division.
@ErikaHolzbaur
@CorySimpsonMD
@AICjanelia
@JLS_Lab
Full-text link:
Cables at work:
Changing up the spatial and temporal scales for this
#FluorescenceFriday
. Here is an overnight acquisition of COS-7 cells stably expressing EGFP-Sec61b to label the endoplasmic reticulum. Time = hh:mm:ss.
Intermediate filaments dressed up in my favorite colormap (🔥glow). This is a Cos7 transfected with mApple-tagged vimentin. I gamma adjusted the movie to visualize the lower density filaments moving out towards the periphery.
More bonus content that didn't make it into
An actin wave bossing around mitochondria. This time without those pesky microtubules (10 uM nocodazole). Maybe I can do a whole "director's cut" on twitter...
Spent the day playing with the AGAVE image viewer from the
@AllenInstitute
. I'm totally blown away - it's now my go-to for volumes. Here are some fun images of the ER in a Cos7 cell.
#FluorescenceFriday
Our perspective on endoplasmic reticulum structure has now been published! In this review, we (
@cjobara
@JLS_Lab
) examine the diverse nanoscale ER structures that have been described by light (see below) and electron microscopy.
GPT4 fixed all of my mediocre imageJ macros. I literally just copy/pasted my code and prompted something like "please make this better" or "please annotate this I don't remember what it does" or "make this work on multichannel images" and it just does it 🤯.
Here's a curious-looking tissue culture cell imaged every minute for an hour and a half. The cell is labeled with a fluorescent marker for the endoplasmic reticulum. In my professional opinion, it looks like a spooky face.
One silver lining of working from home is having the time to browse old data that I had forgotten about. Here are some dynamic actin waves in a zebrafish that I acquired with
@DvirGur
OK, here's my last pitch. Mark your calendars for the Cytoskeletal Ensembles Across Scales subgroup on Monday at
#cellbio2020
. At 3:04 pm (SG15) I'll be talking about mito-actin interactions as well as these actin comet tails driving bursts of mitochondrial movement.
Well that's a nice way to kick off ASCB... my movie of actin dynamics took the top prize in this year's video contest! Thank you judges! If you want to see more actin movies check out the
#cellbio2020
subgroup "Cytoskeletal Ensembles Across Scales" Monday, 12/7 from 1:45-4:45 p.
Congrats to our Image and Video Contest winners! Check out this video of all the top prize winners!
🔗
And also check out this gallery of ALL entries. So hard to choose!
🔗
Read more here:
🔗
Vimentin does some pretty interesting things during mitosis (in some cell types). Lattice light-sheet movie. Vim = orange, MTs = blue.
#notjustcortical
Detecting cell boundaries seemed straightforward, until I applied a square root transformation. You'd be amazed at all the stuff lurking in the depths of your histograms.
For
#FluorescenceFriday
, here is the ER moving at normal speed. The movie was acquired at 31 fps and played back at 31 fps. It really is that dynamic!
Vimentin in a metaphase RPE1 cell. It seems like mitotic intermediate filaments look different in every cell line that I look at. Check out the cool twisting filaments.
I post a lot of light microscopy movies. I love photons, but I guess electrons are pretty great too. Here's a FIB-SEM stack of a U2OS cell with a 3D mito segmentation.
I'm excited to present some of my recent work on ER-vimentin coupling today at
#cellbio2023
. I'll be speaking in the organelle dynamics/quality control joint session in room 254A at 10:05. I can promise lots of live movies (and a bit more info on this 3d rendering)
For reference - in images with high dynamic range (such as this one) it's hard to see very dim signal without saturating brighter pixels. The original image is on the bottom and the square root is on the top.
I call this LUT "BarberPole". It's essentially useless unless you are trying to color code an image based on radial position...and even then it's kind of ambiguous. But I made it and I'm proud.
Made this stable RPE1 line with F-Tractin-mNG a while back and saw all these strange vesicular structures labeled with the probe. Anybody familiar with this artifact or is this an RPE actin thing? TMRM = orange.
For last
#FluoresenceFriday
I shared 1 hour of happy endoplasmic reticulum. For this
#FluorescenceFriday
I present 1 minute of very angry endoplasmic reticulum. Though I guess it really isn't a reticulum anymore 🤔
Should you have 45 minutes to spare tomorrow morning, why not come visit my eposter? I'll be in the Mitochondrial Architecture session talking about actin (and mitochondria).
#cellbio2020
There's something strangely hypnotic about this actin😵💫
GI-TIRF-SIM movie of lifeact-GFP at the very bottom of a dividing cell. Acquired at 1fps, played back at 24fps. Perhaps *slightly* overprocessed.
I finally had the chance to try StayGold (). I was very impressed. 11 slice per frame, 300 total frames, almost no visible bleaching. This is an airyscan movie of TdStayGold-Giantin in a Cos7.