zeiss_apotome
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zeiss_apotome [2025/01/14 11:07] bioimaging [Objectives] |
zeiss_apotome [2025/12/12 11:36] (current) bioimaging |
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| <a href="/facility/bioimaging/lib/exe/fetch.php?media=imager.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=imager.jpg" width=300></a> | <a href="/facility/bioimaging/lib/exe/fetch.php?media=imager.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=imager.jpg" width=300></a> | ||
| <td style="border:0px solid white;"> <p style="line-height:1.8"> | <td style="border:0px solid white;"> <p style="line-height:1.8"> | ||
| - | <b>Location</b>: [Oeiras] Room 0B02<br> | + | <b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=bartolomeu_dias_wing_2025_-_apotome.png">[Oeiras] Room 0B02</a><br> |
| <b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">ZEISS Microscopy</a><br> | <b>Manufacturer</b>: <a href="http://www.zeiss.de/micro" target="_blank">ZEISS Microscopy</a><br> | ||
| <b>Model</b>: <a href="https://www.zeiss.com/microscopy/en/products/light-microscopes/widefield-microscopes/axio-imager-2-for-life-science-research.html">Zeiss Axio Imager 2 with Apotome 2</a><br> | <b>Model</b>: <a href="https://www.zeiss.com/microscopy/en/products/light-microscopes/widefield-microscopes/axio-imager-2-for-life-science-research.html">Zeiss Axio Imager 2 with Apotome 2</a><br> | ||
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| <b>Software</b>: Zen 3.1<br> | <b>Software</b>: Zen 3.1<br> | ||
| <b>Year</b>: 2019<br> | <b>Year</b>: 2019<br> | ||
| - | <b>SN</b>: <br> | + | <b>SN</b>: 3523001603<br> |
|   <br> |   <br> | ||
| Data will be deleted after: <b>1 month</b> | Data will be deleted after: <b>1 month</b> | ||
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| ===== Microscope overview ===== | ===== Microscope overview ===== | ||
| - | The “Zeiss Imager2 + Apotome2'' is a fully automated upright microscope capable of | + | The ZEISS Axio Imager with Apotome 2 is a fully automated upright microscope capable of magnifications from 2.5x to 160x and equipped with two cameras: On the left port a 5MPx CMOS color camera ([[https://www.zeiss.com/microscopy/en/products/cameras/axiocam-105-color.html|Axiocam 105 RGB]], for brightfield/DIC) and on the top port a high sensitivity 4 Mpx monochrome sCMOS camera ([[https://www.biomedical-instruments-trading.com/products/video-devices/568-hamamatsu-orca-flash-4-0-v2|Hamamatsu Flash 4.0v2]], for fluorescence). The system has a Colibri 7 excitation light source with 5 LEDs (see table below). The system’s stage, which can accommodate 8 slides simultaneously, is motorized in XYZ, capable of z-stacks, multiple positions or tile and stitching acquisition (thereby allowing acquisition of large fields of view with several hundred megapixels). |
| - | magnifications from 2.5x to 160x and equipped with two cameras: On the left port a 5MPx CMOS | + | The Imager is also equipped with ApoTome.2, which allows optical sectioning of fluorescent samples using structured illumination. A grid pattern is placed on the illumination path and is projected and shifted sideways; 3 (or more) images are acquired and between each image the grid is moved 1/3 of its cycle. This movement generates images where only the in-focus light is modulated by the grid pattern while out-of-focus light is largely unaffected by the grid. By subtracting pairs of these images, the system removes out-of-focus light and the grid pattern, thus creating an optical section of the sample. |
| - | color camera ([[https://www.zeiss.com/microscopy/en/products/cameras/axiocam-105-color.html|Axiocam 105 RGB]], for brightfield/DIC) and on the top port a high sensitivity 4Mpx | + | |
| - | monochrome sCMOS camera (Hamamatsu Flash 4.0v2, for fluorescence). The system has a | + | |
| - | Colibri7 excitation light source with 5 LEDs (see table below). The system’s stage, which can | + | |
| - | accommodate 8 slides simultaneously, is motorized in XYZ, capable of z-stacks, multiple | + | |
| - | positions or tile & stitching acquisition (thereby allowing acquisition of large fields of view with | + | |
| - | several hundred megapixels!). | + | |
| - | The Imager is also equipped with ApoTome.2, which allows optical sectioning of fluorescent | + | |
| - | samples using structured illumination. A grid pattern is placed on the illumination path and is | + | |
| - | projected and shifted sideways; 3 (or more) images are acquired and between each image the grid | + | |
| - | is moved 1/3 of its cycle. This movement generates images where only the in-focus light is | + | |
| - | modulated by the grid pattern while out-of-focus light is largely unaffected by the grid. By | + | |
| - | subtracting pairs of these images, the system removes out-of-focus light and the grid pattern, thus | + | |
| - | creating an optical section of the sample. | + | |
| ===== System components ===== | ===== System components ===== | ||
| - | ===== Colibri7 excitation lines (LED) ==== | + | ==== LEDs ==== |
| - | ^ Excitation Lines (nm) ^ Abs (W) ^ | + | ^ Line ^ Wavelength ^ Power (with 10x) ^ |
| - | | 385 | 20 | | + | | UV | <fc #be00cc>385 nm</fc> | ~ 160* mW | |
| - | | 475 | 15 | | + | | B | <fc #00c0ff>475 nm</fc> | ~ 120 mW | |
| - | | 555 | 5 | | + | | G | <fc #b3ff00>555 nm</fc> | ~ 20 mW | |
| - | | 590 | 5 | | + | | Y | <fc #ffdf00>590 nm</fc> | ~ 14 mW | |
| - | | 630 | 10 | | + | | R | <fc #ff4f00>630 nm</fc> | ~ 44 mW | |
| + | * don't exceed 5% power as this line bleaches samples quickly. | ||
| - | ==== Emission Filters ==== | + | ==== Filtersets ==== |
| - | ^ Filter name ^ Transmission(nm) ^ Reference ^ | + | ^ Filterset ^ Excitation ^ Dichroic ^ Emission ^ Reference ^ |
| - | | BFP | 450/40 | [[https://www.fpbase.org/spectra/?s=2666,2668,2667,2055&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|96 HE]] | | + | | Blue | 370-410 nm | 420 nm | <fc #0046ff>430-470 nm</fc> | [[https://www.micro-shop.zeiss.com/en/us/shop/filterAssistant/filtersets/489096-9100-000|FS96HE]] | |
| - | | GFP | 425/50 | [[https://www.fpbase.org/spectra/?s=1484,1495,1485,2057&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|38 HE]] | | + | | Green | 450-490 nm | 493 nm | <fc #4aff00>500-550 nm</fc> | [[https://www.micro-shop.zeiss.com/?s=7139069fdc79c&l=en&p=us&f=f&a=v&b=f&id=489038-9901-000&o=|FS38HE]] | |
| - | | Alexa 546 | LP590 | [[https://www.fpbase.org/spectra/?s=1530,1533,1536,2059&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|15]] | | + | | Orange-Red | 540-552 nm | 580 nm | <fc #ffdf00>> 590 nm</fc> | [[https://www.micro-shop.zeiss.com/en/us/shop/filterAssistant/filtersets/488015-0000-000|FS15]] | |
| - | | Cy5 | 690/50 | [[https://www.fpbase.org/spectra/?s=1547,1550,1543,2068&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|50]] | | + | | Red | 625-655 nm | 660 nm | <fc #ff0000>665-715 nm</fc> | [[https://www.micro-shop.zeiss.com/en/us/shop/filterAssistant/filtersets/488050-9901-000|FS50]] | |
| - | | DIC | <800 | - | | + | | DIC | - | - | - | - | |
| - | | mPlum | 629/62 | [[https://www.fpbase.org/spectra/?s=2620,2618,2619,2069&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|64 HE]] | | + | | Orange | 575-600 nm | 605 nm | <fc #ff5300>612-682 nm</fc> | [[https://www.micro-shop.zeiss.com/en/us/shop/filterAssistant/filtersets/489064-0000-000|FS64HE]] | |
| - | | Quad | 425/514/592/709 | [[https://www.fpbase.org/spectra/?s=2657,2601,2602,2658,2577&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|90 HE]] | | + | | QUAD | 370-400 nm | 405 nm | <fc #5400ff>410-440 nm</fc> | [[https://www.micro-shop.zeiss.com/en/us/shop/filterAssistant/filtersets/489090-9100-000| FS90HE]] | |
| ==== Objectives ==== | ==== Objectives ==== | ||
| ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ||
| - | | 2.5x | Plan-Neofluar | air | 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]] | | + | | <fc #c2a446>2.5x</fc> | Plan-Neofluar | air | 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 | Plan-Neofluar | air | 0.3 | 5.5 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420340-9900-000/Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9900-000]] | | + | | <fc #fef042>10x</fc> | Plan-Neofluar | air | 0.3 | 5.5 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420340-9900-000/Objective-EC-Plan-Neofluar-10x-0.30-M27|420340-9900-000]] | |
| - | | 20x | Plan-Apochromat | air | 0.8 | 0.55 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420650-9902-000/Objective-Plan-Apochromat-20x-0.8-M27|420650-9902-000]] | | + | | <fc #0aa05c>20x</fc> | Plan-Apochromat | air | 0.8 | 0.55 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420650-9902-000/Objective-Plan-Apochromat-20x-0.8-M27|420650-9902-000]] | |
| - | | 40x | Plan-Apochromat | air | 0.95 | 0.25 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420660-9970-000/Objective-Plan-Apochromat-40x-0.95-Corr-M27|420660-9970-000]] | | + | | <fc #01b3f5>40x</fc> | Plan-Apochromat | air | 0.95 | 0.25 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420660-9970-000/Objective-Plan-Apochromat-40x-0.95-Corr-M27|420660-9970-000]] | |
| - | | 40x | Plan-Neofluar | oil | 1.3 | 0.21 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420462-9900-000/Objective-EC-Plan-Neofluar-40x-1.30-Oil-DIC-M27|420462-9900-000]] | | + | | <fc #01b3f5>40x</fc> | Plan-Neofluar | oil | 1.3 | 0.21 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420462-9900-000/Objective-EC-Plan-Neofluar-40x-1.30-Oil-DIC-M27|420462-9900-000]] | |
| | 100x | Plan-Apochromat | oil | 1.4 | 0.17 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420792-9900-000/Objective-Plan-Apochromat-100x-1.40-Oil-DIC-M27|420792-9900-000]] | | | 100x | Plan-Apochromat | oil | 1.4 | 0.17 | [[https://www.micro-shop.zeiss.com/en/us/shop/objectives/420792-9900-000/Objective-Plan-Apochromat-100x-1.40-Oil-DIC-M27|420792-9900-000]] | | ||
| ==== Camera ==== | ==== Camera ==== | ||
| + | ^ Model ^ Frame Size ^ Pixel Size (µm) ^ Quantum Efficiency ^ | ||
| + | | [[https://www.biomedical-instruments-trading.com/products/video-devices/568-hamamatsu-orca-flash-4-0-v2|Hamamatsu ORCA-Flash4.0 V2]] | 2048 x 2048 | 13.3 x 13.3 | 70 % | | ||
| + | | [[https://www.zeiss.com/microscopy/en/products/cameras/axiocam-105-color.html|ZEISS Axiocam 105 color]] | 2048 x 2048 | 2.2 x 2.2 | - | | ||
| + | ===== Microscope Turn On Procedure ===== | ||
| + | |||
| + | * Turn on the Power Supply | ||
| + | {{::oeiras_apotome_turningon1.jpg?200|}} | ||
| + | * Turn on the Microscope Body | ||
| + | {{::oeiras_apotome_turningon2.jpg?200|}} | ||
| + | * Turn on the computer (if needed) | ||
| + | * Log In into Windows with your Agendo credentials | ||
| + | * Start the **ZEN** software | ||
| + | |||
| + | ===== Microscope Turn Off Procedure ===== | ||
| + | |||
| + | * Exit the **ZEN** software | ||
| + | * **Log off** the computer | ||
| + | * Turn off the Microscope Body | ||
| + | * Turn off the Power Supply | ||
| ---- | ---- | ||
| [[resources|Back to the Equipment page]] | [[resources|Back to the Equipment page]] | ||
zeiss_apotome.1736849254.txt.gz · Last modified: 2025/01/14 11:07 by bioimaging
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