zeiss_apotome
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zeiss_apotome [2025/01/14 10:13] bioimaging |
zeiss_apotome [2025/01/17 13:09] (current) bioimaging [Camera] |
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| ===== Microscope overview ===== | ===== Microscope overview ===== | ||
| - | GABY WORKING ON IT! | + | 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 ===== | ===== System components ===== | ||
| - | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ | + | ===== 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 ==== | ==== 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]] | |
| + | | 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 ==== | ==== 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]] | [[resources|Back to the Equipment page]] | ||
zeiss_apotome.1736846032.txt.gz · Last modified: 2025/01/14 10:13 by bioimaging
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