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?250}} The Leica STELLARIS STED FLIM is a [[https://www.youtube.com/watch?v=rTkjQ6Q0p5I|point scanning confocal microscope]] designed for advanced fluorescence imaging, super-resolution microscopy and fluorescence lifetime measurements. Built on the [[https://www.leica-microsystems.com/products/confocal-microscopes/p/leica-stellaris/|Leica STELLARIS platform]], it combines a [[https://www.leica-microsystems.com/science-lab/life-science/stellaris-white-light-lasers/|White Light LASER]] (WLL) excitation source,  an Acousto-Optical Beam Splitter (AOBS), highly sensitive [[https://www.leica-microsystems.com/science-lab/life-science/power-hyd-detectors/|Power HyD detectors]] and prism-based spectral detection to provide flexible excitation of up to 8 simultaneous laser lines from 440 to 790 nm. When combined with the Power HyD R detector, you can detect fluorophores in the near infrared range (up to 850 nm) such as Cy7. It is also equipped with a 405 nm diode laser, a motorized XY scanning stage, a SuperZ Galvo Stage for rapid Z acquisition, and an Adaptive Focus Control (AFC) closed-loop hardware autofocus system for long-term stability during time-lapse experiments. A large environmental enclosure with temperature and CO2 control enables live-cell imaging under physiological conditions. The confocal scanhead incorporates a field-of-view scanner with galvanometric mirrors optimized for high resolution and sensitivity and includes a Power HyD X detector optimized for [[https://www.leica-microsystems.com/science-lab/life-science/a-guide-to-fluorescence-lifetime-imaging-microscopy-flim/|fluorescence lifetime applications]]. For super-resolution imaging, the system is equipped with STED technology, including pulsed 589 nm and 775 nm depletion lasers, enabling imaging with resolution well below the diffraction limit. The 3D STED module provides tunable super-resolution in X, Y and Z, while [[https://www.leica-microsystems.com/science-lab/life-science/extended-live-cell-imaging-at-nanoscale-resolution/|TauSTED Xtend]] combines STED and fluorescence lifetime information to improve image quality, contrast and resolution at lower light doses, making it particularly suitable for live-cell super-resolution imaging. The system supports multicolour STED imaging across much of the visible spectrum and facilitates the investigation of molecular organization, interactions and nanoscale cellular structures. The microscope also includes [[https://www.leica-microsystems.com/products/confocal-microscopes/p/stellaris-8-falcon/|STELLARIS FALCON (FAst Lifetime CONtrast)]], providing fluorescence lifetime imaging microscopy (FLIM) at confocal imaging speeds. Fully integrated within LAS X software, FALCON supports exponential fitting, [[https://www.leica-microsystems.com/science-lab/life-science/phasor-analysis-for-flim-fluorescence-lifetime-imaging-microscopy/|FLIM phasor analysis]], pattern fitting and [[https://www.leica-microsystems.com/science-lab/life-science/what-is-fret-with-flim/|FLIM-FRET analysis]], enabling quantitative studies of molecular interactions, environmental sensing and dynamic cellular processes.

  * **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, and Power HyD R) 
  * **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 #0000ff>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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