Luis AJ Alvarez

Betty Yuen-Torres and Luis Alvarez holding the AACR SpectraPlex poster

Habemus Poster 🙃 #AACR2025
Just got the printout for our SpectraPlex Poster - it is proper size for the data we wanted to fit 😇. Come by tomorrow - section 7 board 24 poster 173 | happy to chat 3D high multiplexing imaging in cancer. Session starts at 14:00.

#AACR25 sign in Chicago for the annual American Association Of Cancer Reseaech conference

@ Chicago for #AACR2025 - if you are around and want to talk microscopy just drop me a message to meet up. Also we got a running STELLARIS with SpectraPlex at the @leicamicrosystems.bsky.social booth for 15+ channels on one go at the booth

📣 The application deadline for #EMBLFluorescenceLifetime has been extended until 28 Apr! ✍🏼

Are you a PhD student or a postdoc with experience in fluorescence microscopy and want to enhance your skills in confocal fluorescence microscopy and lifetime-based readouts? 🔬

➡️s.embl.org/lef25-01-bl

Cervical cancer tissue stained with PanCK-Opal 780 - Imaged on a Leica STELLARIS FALCON

Cervical Cancer tissue - PanCK Opal780 stained - left intensity only | right Fluorescence lifetime (rainbow 0,5-1,4 ns) showing the PanCK on cyan, non specific signal blue endogenous tissue signals in red #FluorescenceFriday #FluorescenceLifetime - interested ? FLIM course in July @ EMBL 😜

We have an amazing group of speakers and hands on sessions around Fluorescence Lifetime Imaging (multiphoton, metabolism, STED-FLIM, biosensing, Cryo, Plant imaging, multiplexing, FRET, functional dynamics and FLIM analysis.
Follow the link for program & application !

📣Application deadline for #EMBLFluorescenceLifetime is approaching ⏰

Do you have experience in fluorescence microscopy and want to enhance your skills in confocal fluorescence microscopy and in lifetime-based readouts?

Then apply for this hands-on course by 21 April: ➡️s.embl.org/lef25-01-bl

3D rendering of a Zebrafish embryo showing vasculature (GFP - magenta) and endogenous fluorescence (cyan) separated through Fluorescence Lifetime Imaging

3D rendering of a Zebrafish embryo showing vasculature (GFP) and endogenous fluorescence

#fluorescencefriday
#fluorescencelifetime
3D rendering Zebrafish embryo vasculature + endogenous fluorescence - intensity only (greyscale) corresponding lifetime separated (GFP - magenta | endogenous - cyan)
Interested? apply for our summer course @ EMBL in Heidelberg: www.embl.org/about/info/c...

2/2 hence encoding data is a 2 channel process in tiff. A workaround is to set it in HSV colour space with the brightness value set for intensity and playing on hue and saturation for the colour. I can provide raw images but it might require additional work to sort out LUTs.

1/2 The lifetime bit is represented on a multicolour gradient. With the standard LUT being blue|cyan|green|yellow|red. It codes for intensity + lifetime or average arrival time (AAT) on a single channel. Values from 0 to 3/4 with 2 decimals (ns) or 3 digits (ps)

This has been my experience as well and for traditional 3-4 colour images it is always possible to find something fine when you hit 6+ and are integrating them I have not seen options that make sense - still keeping an eye out just in case 🙃.

Application Note: TauSense: the potential of STELLARIS TauSense from Leica Microsystems is a new way to use lifetime-based information for enhanced multiplexing and functional imaging

For the classical fluorescence lifetime rainbow LUT please have a look at fig 1b here: www.nature.com/articles/d42...

Application Note: 3D high-multiplex imaging in cancer immunology 3D multiplexed imaging in cancer immunology enables a better characterization of tissue changes and cell interactions. Here, we introduce a one-shot, high-multiplexing workflow on the STELLARIS confoc...

For the 15 colours please look at fig 3 a here : www.nature.com/articles/d42...

We tend to show subsets of markers at a time as well. Fig 3 b-g. But also fig 2 here www.nature.com/articles/d42...

Very nice ! We had some requests about colour blind alternatives for the “rainbow” LUT used for FLIM or to get multiple FLIM channels. I could not find a solution. Now that we can get 15 markers at once I anticipate similar question for colour blind friendly set. Any ideas ? 🙈

Pithiviers pour galette des rois pour l’épiphanie | Pithiviers puffy pastry French cake for wise man for January sixth

Part de galette des rois - Chocolat + frangipane filling

À chocolate & Frangipane filled Pithiviers for Galette des rois for Heilige Drei Könige | l’épiphanie | best way to close the Christmas season 😋🤩. I hope your end of the year a blast.

Decorated and lighted up Christmas tree with gifts below

35 min past midnight and every thing is set for the kids to wake up early on Christmas Day 🤣. Happy holidays to everyone and I hope you have a great time with family and friends!

Calling all #SciComm people who specialise in scientific animations. I would like to talk to someone about making some animations to communicate our research in single-molecule biophysics. Please message me if you are interested and share with your networks! 🧪

15 colour beads - 0.7-0.9 μm | 405-850 nm | 50 X 75 μm image | xy view of a z stack

15 colour beads from 405 to 850 nm | side view of the z-stack with a line through the middle showing optimal chromatic correction across the complete spectra. All beads perfectly centered on the focal plane

15 colour beads 3D image | 2 μm z view showing all beads from 405 to 850 nm on the same z plane with close to isometric resolution

15 colours beads on a single slide 🙃. Fluorescence emissions from 410 to 850 nm. 0.7-0.9 μm beads | z-stack showing the side view with proper chromatic correction across all wavelengths! Imaged on a STELLARIS with an extended white light laser (WLL) and SpectraPlex #FluorescenceFriday

How does it look like when calibrating it expresses in bacteria ? W/o purification ? & a calibration expresses in mammalian cells ?