GIMM Bioimaging Wiki

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<b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=Bartolomeu_Dias_Wing_2025_-_STELLARIS.png">[Oeiras] Room 0B02</a><br> 
<b>Manufacturer</b>: <a href="https://www.leica-microsystems.com/" target="_blank">Leica Microsystems</a><br>
<b>Model</b>: <a href="https://www.leica-microsystems.com/products/confocal-microscopes/p/stellaris-sted/">Leica STELLARIS STED</a><br>
<b>Nickname</b>: "STELLARIS"<br>
<b>Software</b>: LAS X<br>
<b>Year</b>: 2026 <br>
<b>SN</b>: 840000544 <br>
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Data will be deleted after: <b>1 month</b>
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&rarr; <a href="/facility/bioimaging/doku.php?id=leica_stellaris_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Leica STELLARIS STED FLIM Quality Control</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=leica_stellaris_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  Leica STELLARIS STED FLIM Usage Statistics</a>
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===== Microscope overview =====

{{  :sted_principle.png?240}} The Leica STELLARIS STED FLIM is a [[https://www.youtube.com/watch?v=rTkjQ6Q0p5I|point scanning confocal microscope]] designed for advanced fluorescence imaging, [[https://www.leica-microsystems.com/science-lab/life-science/a-guide-to-fluorescence-lifetime-imaging-microscopy-flim/|fluorescence lifetime imaging (FLIM)]] and super-resolution microscopy. Built on the [[https://www.leica-microsystems.com/products/confocal-microscopes/p/leica-stellaris/|Leica STELLARIS platform]], it features a [[https://www.leica-microsystems.com/science-lab/life-science/stellaris-white-light-lasers/|White Light LASER]] (440-790 nm) and 405 nm laser as excitation sources, AOBS beam splitting, highly sensitive [[https://www.leica-microsystems.com/science-lab/life-science/power-hyd-detectors/|Power HyD detectors]] and spectral detection for flexible multicolor imaging, including near-infrared fluorophores up to 850 nm, such as Cy7. The system is optimized for live-cell imaging, featuring a motorized stage, SuperZ Galvo Stage for rapid Z acquisition, hardware autofocus (AFC) and a temperature- and CO₂-controlled Okolab dark enclosure for live-cell imaging in physiological conditions. FLIM capabilities are provided by the Power HyD X detector and [[https://www.leica-microsystems.com/products/confocal-microscopes/p/stellaris-8-falcon/|STELLARIS FALCON (FAst Lifetime CONtrast)]], enabling fast lifetime imaging, [[https://www.leica-microsystems.com/science-lab/life-science/what-is-fret-with-flim/|FLIM-FRET experiments]], [[https://www.leica-microsystems.com/science-lab/life-science/phasor-analysis-for-flim-fluorescence-lifetime-imaging-microscopy/|phasor analysis]] and [[https://www.leica-microsystems.com/science-lab/life-science/tausense-technology-imaging-tools/|TauSense]] lifetime-based contrast and multiplexing tools.
For super-resolution imaging, the system is equipped with [[https://www.leica-microsystems.com/science-lab/life-science/a-guide-to-super-resolution/|2D and 3D STED technology]] using pulsed 589 nm and 775 nm depletion lasers, providing resolution well below the diffraction limit. [[https://www.leica-microsystems.com/science-lab/life-science/extended-live-cell-imaging-at-nanoscale-resolution/|TauSTED Xtend]] combines STED and lifetime information to improve image quality and resolution while reducing light exposure, making the system particularly suitable for live-cell nanoscale imaging. 

  * **Microscope**: [[https://www.leica-microsystems.com/products/light-microscopes/p/dmi8/|Leica DMi8 CS inverted]]
  * **LED Illumination**: [[https://www.coolled.com/products/pe-300white/|CoolLED pE-300white]]
  * **Detectors**: [[https://www.leica-microsystems.com/science-lab/life-science/power-hyd-detectors/|Power hybrid detectors]] 
    * Power HyD S
    * Power HyD X for FLIM imaging
    * Power HyD R for near-infrared imaging  
  * **Features**
    * |  [[https://www.leica-microsystems.com/science-lab/life-science/a-guide-to-super-resolution/|2D and 3D Super Resolution STED]]
    * | [[https://www.leica-microsystems.com/science-lab/life-science/extended-live-cell-imaging-at-nanoscale-resolution/|TauSTED Xtend]]
    * | [[https://www.leica-microsystems.com/science-lab/life-science/tausense-technology-imaging-tools/|TauSense]]
    * | [[https://www.leica-microsystems.com/products/confocal-microscopes/p/stellaris-8-falcon/|FALCON FLIM Imaging]]
    * | [[https://www.leica-microsystems.com/science-lab/life-science/obtain-maximum-information-from-your-specimen-with-lightning/|LIGHTNING adaptive deconvolution]]
    * | [[https://youtu.be/3uq42XefHhY?si=EWUFLyqk2z3Q1HG3|Adaptive Focus Control (AFC)]]

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{{:warning.png?25|}} Data files older than **1 month** will be automatically deleted on this system, please copy your data to the GIMM server using the desktop link.

===== Additional information for sample preparation =====

    * [[https://downloads.leica-microsystems.com/STELLARIS%208%20STED/Application%20Note/STELLARIS%20STED%20Sample%20Preparation%20Guide.pdf|The Guide to STED Sample Preparation (application report)]] 

===== System components =====

==== LASERs ====

^  STED LASER  ^  Wavelength  ^
|  STED 589  |  <fc #ffe200>589 nm</fc>  |
|  STED 775  |  <fc #be0000>775 nm</fc>  |

^  LASER  ^  Wavelength  ^
|  Violet  |  <fc #a600ff>405 nm</fc>  |
|  WLL  |  <fc #0071ff>440 nm</fc> - <fc #be0000>790 nm</fc>  |

==== Filterset (for eyepieces) ====

^  Filterset  ^  Excitation  ^  Dichroic  ^  Emission  ^  Reference  ^
|  Blue  |  375-435 nm  |  455 nm  |  <fc #00a9ff>450-490 nm</fc>  |  [[https://www.fpbase.org/spectra/?s=6094,6092,6093&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|LED 405 (15525338)]]  |
|  Green  |  450-490 nm  |  495 nm  |  <fc #4aff00>500-550 nm</fc>  |  GFP (15525314)  |
|  Orange-Red  |  518-562 nm  |  570 nm  |  <fc #ff7700>>590 nm</fc>  |  RHOD LP (15525303)  |

==== Objectives ====

^  Magnification   ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  Reference  ^
|  <fc #fef042>10x</fc>  |  HC PL APO CS2  |  Air  |  0.40  |  2.56  |  [[https://www.leica-microsystems.com/objectivefinder/detail/objective/506424/|11506424]]  |
|  <fc #0aa05c>20x</fc>  |  HC PL APO IMM CORR CS2  |  Water  |  0.75  |  0.66  |  [[https://www.leica-microsystems.com/objectivefinder/detail/objective/506343/|11506343]]  |
|  <fc #01b3f5>40x</fc>  |  HC PL APO Oil CS2  |  Oil  |  1.30  |  0.17  |  [[https://www.leica-microsystems.com/objectivefinder/detail/objective/506428/|11506428]]  |
|  <fc #2c80c0>63x</fc>  |  HC PL APO Oil CS2  |  Oil  |  1.40  |  0.14  |  [[https://www.leica-microsystems.com/objectivefinder/detail/objective/506350/|11506350]]  |
|  100x  |  HC PL APO OIL STED WHITE  |  Oil  |  1.40  |  0.13  |  [[https://www.leica-microsystems.com/objectivefinder/detail/objective/506378/|11506378]]  |

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