Location: Room P2-A-24 ( 47219)
Manufacturer: ZEISS Microscopy
Model: Axio Observer
Nickname: "Cell Observer"
Software: ZEN 3.0 (blue edition)
Year: 2015
SN: 3851002396
 
Data will be deleted after: 15 days  
 
→ Zeiss Cell Observer Quality Control
→ Zeiss Cell Observer Usage Statistics

===== Microscope overview ===== {{ :widefield_lightpath.png?0x300}}The Zeiss Cell Observer is a fully motorized inverted //widefield fluorescence microscope// ideal for live-cell imaging with fast acquisition speeds. It is equipped with a large cage incubator and a stage top incubator for temperature control and CO2 supply and with Definite Focus for hardware focus control during long time-lapse acquisitions. Using the ZEN software you can control the motorized filters, shutters and stage to simultaneously set up multi-color time-lapse imaging experiments in multiple stage positions. The stage is furthermore equipped with a piezo for fast Z displacement. You can choose between a HXP metal halide lamp or fast-switching LEDs for illumination. Its cooled [[https://asset-downloads.zeiss.com/catalogs/download/mic/a8e5b4d7-25f6-4069-8f03-c6713d044c87/EN_product-info_Axiocam-506_mono.pdf|Axiocam 506m]] camera allows you to capture weak fluorescent signals and minimize photobleaching/photodamage in light sensitive samples. It is also equipped with a high sensitivity [[https://www.photometrics.com/products/prime-family/primebsiexpress|Photometrics Prime BSI Express]] sCMOS camera for high resolution imaging with maximized signal detection. * **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]] * **Camera** (left port): [[https://asset-downloads.zeiss.com/catalogs/download/mic/a8e5b4d7-25f6-4069-8f03-c6713d044c87/EN_product-info_Axiocam-506_mono.pdf|Axiocam 506 mono CCD]] * **Camera** (right port): [[https://www.photometrics.com/products/prime-family/primebsiexpress|Photometrics Prime BSI Express sCMOS]] {{::phenocycler.png?260 |}}This system is also set up to work together with the [[https://www.youtube.com/watch?v=OQGJX45Wsgw|AKOYA PhenoCycler]] fluidics instrument for automated iterative imaging cycles of whole slide tissues for [[https://www.akoyabio.com/why-spatial-biology/|Spatial Biology]] applications, such as image-based quantification of biomarkers to predict immunotherapy response, or quantification of cells by lineage and variant with single-cell resolution in the context of an intact tissue sample. * [[https://www.youtube.com/watch?v=OQGJX45Wsgw|Understanding the PhenoCycler Technology]] * [[https://www.akoyabio.com/webinar/a-beginners-guide-to-codex/|A Beginner’s Guide to CODEX]] * [[https://www.akoyabio.com/webinar/spatial-phenotyping-a-revolutionary-approach-to-biomarker-discovery-for-cancer-immunotherapy/|Spatial Phenotyping: A Revolutionary Approach to Biomarker Discovery for Cancer Immunotherapy]] {{:warning.png?25|}} Data files older than **15 days** will be automatically deleted on this system, please copy your data to the iMM server using the desktop link. ===== Booking Rules ===== - **Users** can book at maximum **8 hours per week** - This usage restriction does not apply for **weekends** and for working days **before 9:00** and **after 21:00** - Exceptions to these rules require **approval from [[joserino@medicina.ulisboa.pt|José Rino]]**. ===== System components ===== ==== LEDs ==== ^ Line ^ Wavelength ^ | UV | 385 nm | | B | 475 nm | | G | 555 nm | | Y | 590 nm | | R | 630 nm | ==== Filtersets ==== ^ Position ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ | 1 | HE GFP | [[https://www.micro-shop.zeiss.com/?s=23108437692794b&l=en&p=us&f=f&a=v&b=f&id=489038-9901-000&o=|FS38HE]] | 450-490 nm | 495 nm | 500-550 nm | | 2 | HE DsRed | [[https://www.micro-shop.zeiss.com/?s=23108437692794b&l=en&p=us&f=f&a=v&b=f&id=489043-9901-000&o=|FS43HE]] | 538-562 nm | 570 nm | 570-640 nm | | 3 | Cy5 | [[https://www.micro-shop.zeiss.com/?s=23108437692794b&l=en&p=us&f=f&a=v&b=f&id=488050-9901-000&o=|FS50]] | 625-655 nm | 660 nm | 665-715 nm | | 4 | Analyzer DIC ||||| | 5 | QUAD | [[https://www.micro-shop.zeiss.com/en/us/shop/filterAssistant/filtersets/489090-9100-000|FS90HE]] | 370-400 nm | 405 nm | 410-440 nm | | ::: | ::: | ::: | 450-488 nm | 493 nm | 499-529 nm | | ::: | ::: | ::: | 540-570 nm | 575 nm | 580-605 nm | | ::: | ::: | ::: | 615-648 nm | 653 nm | 659-759 nm | | 6 | DAPI | [[https://www.micro-shop.zeiss.com/?s=23108437692794b&l=en&p=us&f=f&a=v&b=f&id=488049-9901-000&o=|FS49]] | 335-383 nm | 395 nm | 420-470 nm | ==== Objectives ==== ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | 10x | EC Plan-Neofluar Ph1 | Air | 0.30 | 5.20 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420341-9911-000/Objective-EC-Plan-Neofluar-10x-0.30-Ph1-M27|420341-9911-000]] | | 20x | Plan-Apochromat | Air | 0.80 | 0.55 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420650-9901-000/Objective-Plan-Apochromat-20x-0.8-M27|420650-9901-000]] | | 20x | EC Plan-Neofluar Ph 2 | Air | 0.50 | 2.00 | [[https://www.micro shop.zeiss.com/en/de/shop/objectives/440341-9904-000/Objective-EC-Plan-Neofluar-20x-0.5-Ph2|440341-9904-000]] | | 40x | EC Plan-NeoFluar | Air | 0.75 | 0.71 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420360-9900-000/Objective-EC-Plan-Neofluar-40x-0.75-M27|420360-9900-000]] | | 63x | Plan-Apochromat DIC | Oil | 1.40 | 0.19 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420782-9900-000/Objective-Plan-Apochromat-63x-1.4-Oil-DIC-M27|420782-9900-000]] | | 63x | Plan-Apochromat Ph3 | Oil | 1.40 | 0.19 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420781-9910-000/Objective-Plan-Apochromat-63-1.4-Oil-Ph3-M27|420781-9910-000]] | **Upon request:** ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | 1.25x | EC Epiplan-Neofluar | Air | 0.035 | 4.00 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/422310-9900-000/Objective-EC-Epiplan-Neofluar-1.25x-0.03-M27|422310-9900-000]] | | 5x | EC Plan-NeoFluar | Air | 0.16 | 18.50 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420330-9901-000/Objective-EC-Plan-Neofluar-5x-0.16-M27|420330-9901-000]] | | 10x | EC Plan-Neofluar | Air | 0.30 | 5.20 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420340-9901-000/Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9901-000]] | | 25x | LCI Plan-Neofluar Corr DIC | Oil/Glyc/W | 0.80 | 0.21 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420852-9973-000/Objective-LCI-Plan-Neofluar-25x-0.8-Imm-Corr-DIC-M27|420852-9972-000]] | | 40x | Plan-Apochromat Corr | Air | 0.95 | 0.25 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/420660-9970-000/Objective-Plan-Apochromat-40x-0.95-Corr-M27|420660-9970-000]] | | 40x | Plan-Neofluar Ph3 | Oil | 1.30 | 0.20 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440451-9903-000/Objective-EC-Plan-Neofluar-40x-1.30-Oil-Ph3|440451-9903-000]] | | 100x | Plan-Apochromat DIC | Oil | 1.40 | 0.17 | [[https://www.micro-shop.zeiss.com/en/de/shop/objectives/440782-9902-000/Objective-Plan-Apochromat-100x-1.40-Oil-DIC|440782-9902-000]] | ==== Camera ==== ^ Model ^ Type ^ Frame Size ^ Pixel Size (µm) ^ | [[https://www.zeiss.com/microscopy/int/products/microscope-cameras/axiocam-506-mono.html|Axiocam 506 mono]] | CCD | 2752 x 2208 | 4.54 x 4.54 | ===== System Turn On Procedures ===== * Turn on the HXP 120 fluorescent light source (if used) {{zeiss_axio_observer_hxp.jpg?120|}} * Turn on the temperature and CO2 controller - white power strip (if you need them). * If you need CO2, open the Cell Observer CO2 valve * Turn on the motorized stage controller (**1** on remote control) {{zeiss_axio_observer_a.jpg?height=150|}} * Turn on the microscope power supply (**2** on remote control) {{zeiss_axio_observer_b.jpg?height=150|}} * Turn on the computer * Login in to Windows with your Agendo credentials * Start the **ZEN** software * Select HXP or LED Workspace in ZEN (top left) {{::workspace_selection.png?250|}} ===== System Turn Off procedures ===== If there is another user for this microscope is the __next hour__: * Leave the fluorescent light source on * Clean up immersion objectives Else: * Exit the ZEN software * Turn off the computer (not the UPS!) * Clean up immersion objectives * Turn off the microscope power supply (**2** on remote control) * Turn off the motorized stage controller (**1** on remote control) * Turn off the Temperature and CO2 controller - white power strip (if used) * Close the Cell Observer CO2 valve (if used) * Turn off the HXP 120 fluorescent light source (if used) ---- [[resources|Back to the Equipment page]]