Location: [Oeiras] Room 0B02
Manufacturer: ZEISS Microscopy
Model: Zeiss Axio Imager 2 with Apotome 2
Nickname: Apotome
Software: Zen 3.1
Year: 2019
SN:
 
Data will be deleted after: 1 month  
 
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===== Microscope overview ===== The “Zeiss Imager2 + Apotome2" 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 4Mpx 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 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 ===== ===== Colibri7 excitation lines (LED) ==== ^ Excitation Lines (nm) ^ Power (mW; @ 10x obj) ^ | 385 | ~160* | | 475 | ~120 | | 555 | ~20 | | 590 | ~14 | | 630 | ~44 | * don't exceed 5% power as this line bleaches samples quickly. ==== Emission Filters ==== ^ Filter name ^ Transmission(nm) ^ 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]] | | 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]] | | 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]] | | 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]] | | DIC | <800 | - | | 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]] | | 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]] | ==== Objectives ==== ^ 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]] | | 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]] | | 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]] | | 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]] | | 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]] | | 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 ==== ^ 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?300|}} * Turn on the ''Microscope Body'' {{::oeiras_apotome_turningon2.jpg?300|}} * Turn on the computer * Log In into Windows with your Agendo credentials * Start the **Zen** software ===== Microscope Turn Off Procedure ===== * Exit the **Zen** software * Log off from the computer * Turn off the ''Microscope Body'' * Turn off the ''Power Supply'' ---- [[resources|Back to the Equipment page]]