Location: [Oeiras] Room 0B06
Manufacturer: ZEISS Microscopy
Model: LSM 980 with Airyscan 2
Nickname: "980"
Software: ZEN 3.12
Year: 2019
SN: 2871000107
 
Data will be deleted after: 1 month  
 
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===== Microscope overview ===== {{ :confocal_pinhole.png?340}} The Zeiss LSM 980 with Airyscan 2 is a [[https://www.youtube.com/watch?v=rTkjQ6Q0p5I|point scanning confocal microscope]] is an inverted microscope equipped with a laser scanning head with a total of 6 PMT/GaAsp + 1 Airyscan detectors. The Airyscan can achieve non-diffraction limited optical sectioning with resolutions down to 140nm laterally and ~400nm axially. It also allows acquisition in “Multiplexing” mode(s), which accelerate the acquisition speed (4/8x faster) or in sensitivity mode that enhances the signal-to-noise acquisition. The microscope is also equipped with a dark heating and atmospheric control chamber to allow live imaging of mammalian cells (O2 control is also possible, upon request). We have available an objective inverter for some intra-vital or open-dish applications (upon request only, requires preparation). * **Microscope**: [[https://www.zeiss.com/microscopy/en/products/light-microscopes/widefield-microscopes/axio-observer-for-life-science-research.html|Zeiss Axio Observer]] * **LED Illumination**: [[https://www.zeiss.com/microscopy/en/products/accessories/colibri.html|Zeiss Colibri 7]] * **Features** * | [[https://www.imdik.pan.pl/images/Us%C5%82ugi_dokumenty/Przydatne_informacje/2d_Definite_Focus_information.pdf|Definite Focus.3]] * | [[https://www.youtube.com/watch?v=tUiEa7vcggQ|Airyscan 2 Multiplex]] * | [[https://www.zeiss.com/microscopy/en/about-us/newsroom/press-releases/2021/zeiss-lsm-plus-and-zeiss-airyscan-joint-deconvolution.html|LSM Plus]] * | [[https://www.zeiss.com/microscopy/en/about-us/newsroom/press-releases/2021/zeiss-lsm-plus-and-zeiss-airyscan-joint-deconvolution.html|Joint Deconvolution]]

* [[https://www.youtube.com/watch?v=zA4xqq-EL9g|ZEISS Webinar: LSM 980 Airyscan 2 Multiplex mode]] {{: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. ===== System components ===== ==== Lasers ===== ^ Laser ^ Excitation line ^ Maximum Power ^ Description (use) ^ | 405 nm (violet) | 405 nm | 15 mW | DAPI, Hoechst | | 488 nm (blue) | 488 nm | 13 mW | GFP, FITC, Alexa488 | | 561 nm (green) | 561 nm | 13 mW | RFP, TRITC, Cy3, Dylight Orange | | 639 nm (red) | 639 nm | 10 mW | Cy5, APC, Alexa/ATTO633-647 | If you require more laser lines (e.g., 445nm - CFP/Cerulean, 514nm - YFP, or 594nm mCherry/AF594), we suggest as an alternative the [[https://imm.medicina.ulisboa.pt/facility/bioimaging/doku.php?id=zeiss_lsm_980-32|ZEISS LSM 980-32]] in Lisbon. ==== Objectives ==== ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | 2.5x | | - | 0.085 | 8.8 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420320-9902-000/Objective-EC-Plan-Neofluar-2.5x-0.085-M27|420320-9902-000]] | | 10x | EC Plan-Neofluar | Air | 0.45 | 2.0 | [[https://www.micro-shop.zeiss.com/en/ee/shop/objectives/420640-9900-000/Objective-Plan-Apochromat-10x-0.45-M27|420640-9900-000]] | | 20x | Plan-Apochromat | Air | 0.75 | 0.55 | [[https://www.micro-shop.zeiss.com/en/ee/shop/objectives/420150-9900-000/Objective-Fluar-20x-0.75-M27|420150-9900-000]] | | 25x | | W/Sil/Gly/Oil* | 0.80 | 0.57 | [[https://www.micro-shop.zeiss.com/en/ee/shop/objectives/420852-9870-790/Objective-LD-LCI-Plan-Apochromat-25x-0.8-Imm-autocorr-DIC-M27|420852-9870-790]] | | 40x | LD C-Apochromat Corr | Water | 1.10 | 0.62 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/421867-9970-000/Objective-LD-C-Apochromat-40x-1.1-W-Corr-M27|421867-9970-000]] | | 63x | Plan-Apochromat DIC | Oil | 1.40 | 0.19 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420782-9900-799/Objective-Plan-Apochromat-63x-1.4-Oil-DIC-M27|420782-9900-799]] | *ask staff for help setting up the proper immersion medium setting ==== Detector(s) ==== 1x blue-green multialkali PMT (15-20% QE) 4x GaAsP PMT spectral detectors (tunable between 450-750 nm) (~45% QE) 1x green-red multialkali PMT (10-25% QE) 1x T-PMT detector (brighfield) 1x Airyscan2 detector (~45% QE after processing Airyscan data) ==== Airyscan2 emission filters ===== ^ Position ^ Emission ^ | 1 | Plate | | 2 | BP 420 - 480 + BP 495 - 550 | | 3 | BP 420 - 470 + BP 570 - 630 | | 4 | BP 420 - 500 + LP 605 | | 5 | BP 465 - 505 + BP 525 - 585 | | 6 | BP 495 - 550 + BP 570 - 630 | | 7 | BP 495 - 555 + LP 660 | | 8 | BP 570 - 620 + LP 660 | ==== Eyepiece filterset(s) (COLIBRI LED illumination) ===== ^ Position ^ Transmission ^ Description (use) ^ Excitation ^ | DAPI | 445/50 nm | DAPI, CFP or Hoechst filter | 385nm | | GFP | 525/50 nm | RFP or TRITC (LongPass) | 493nm | | RFP | LP590 nm | GFP or FITC filter | 555nm | | Mirror | - | Mirror for LSM | - | ===== How to start your session ===== The system should be always on, you only need to: * Turn on HXP (if needed) {{::Oeiras_LSM980_hxp.png?100|}} * If incubation is needed (turn on 30 mins before acquiring): * Turn the microscope body off {{::Oeiras_LSM980_microscope_body.png?150|}} * Turn the incubation controller on {{::Oeiras_LSM980_incubation.png?100|}} * Turn the microscope body on * Log into Windows with your Agendo credentials * Start **ZEN** ===== How to end your session ===== If there is another user for this microscope in the __next hour__: * Exit ZEN, leave the lasers on * Log off the computer * Clean up immersion objectives (gently wipe with optic cleaning paper and EtOH embedded paper) Else: * Turn off the lasers in **ZEN** * Clean up immersion objectives (gently wipe with optic cleaning paper and EtOH embedded paper) * Turn off HXP (if used) * Turn off incubation controller (if used) * Exit the ZEN software * Log off the computer ===== Microscope Turn On Procedure (after complete shutdown) ===== If the system is completely off, please follow this procedure: * Turn on the ''Microscope power supply'' * Turn on the ''Laser key'' * Turn on ''HXP'' (if needed) * If ''incubation'' is needed (turn on 30 mins before acquiring): * Turn the microscope body off * Turn the incubation controller on * Turn the microscope body on * Turn on the ''computer'' * Log into Windows with your Agendo credentials * Start **ZEN** ===== Microscope Turn Off Procedure (for complete shutdown) ===== Please only shutdown the system if instructed to do so by the Bioimaging Staff. * Turn off the lasers in **ZEN** * Exit ZEN * Shutdown the computer * Turn off the ''Laser key'' * Turn off HXP (if used) * Turn off the ''Microscope power supply'' ---- [[resources|Back to the Equipment page]]