oeiras_3i_marianas_sdc
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oeiras_3i_marianas_sdc [2025/01/07 16:09] bioimaging [Lasers] |
oeiras_3i_marianas_sdc [2025/01/08 17:36] (current) bioimaging |
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| ===== Microscope overview ===== | ===== Microscope overview ===== | ||
| - | {{ :yokogawa_sd.png?350|}} The 3i Marianas SDC //spinning disk confocal microscope// is a fast imaging system which provides a trade-off between confocality, resolution and speed, well suited for the observation of dynamics in living cells and small organisms. The Marianas SDC provides fast acquisition (up to ~100fps, with limited ROI, ~30FPS full frame), and with lower phototoxicity and bleaching. A spinning disk with multiple small pinholes is installed between the light source and specimen to generate point-like illumination covering the whole area “simultaneously”. Each small aperture serves as a detecting pinhole to remove out-of-focus light. | + | {{ :yokogawa_sd.png?350|}} The 3i Marianas SDC //spinning disk confocal microscope// is a fast imaging system which provides a combination of confocality, resolution and speed, well suited for the observation of dynamics in living cells and small organisms. The Marianas SDC provides fast acquisition (up to ~100fps, with limited ROI, ~30FPS full frame), and with lower phototoxicity and bleaching. A spinning disk with multiple small pinholes is installed between the light source and specimen to generate point-like illumination covering the whole area “simultaneously”. Each small aperture serves as a detecting pinhole to remove out-of-focus light. |
| This system was built to be versatile, and allow imaging of different sample sizes. It includes both high and low magnification objectives (which require different pinhole sizes, 25 or 50um), DIC, a removable heating stage with CO2 delivery, and photomanipulation modules (Phasor and Ablate!). The scanhead allows 1x or 2x magnification (for extra magnification and to allow Nyquist sampling and deconvolution). | This system was built to be versatile, and allow imaging of different sample sizes. It includes both high and low magnification objectives (which require different pinhole sizes, 25 or 50um), DIC, a removable heating stage with CO2 delivery, and photomanipulation modules (Phasor and Ablate!). The scanhead allows 1x or 2x magnification (for extra magnification and to allow Nyquist sampling and deconvolution). | ||
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| | 594 | 594 nm | 100 mW | | | 594 | 594 nm | 100 mW | | ||
| | 637 | 637 nm | 140 mW | | | 637 | 637 nm | 140 mW | | ||
| + | | Ablate! | 355 nm | 60 μJ/200 Hz | | ||
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| ^ Position ^ Filter name ^ Transmission (nm) ^ Dichroic ^ Reference ^ | ^ Position ^ Filter name ^ Transmission (nm) ^ Dichroic ^ Reference ^ | ||
| - | | | | | | | | | + | | 1 | Quad (c###q) | 440/40 521/21 607/34 700/45 | 405/488/561/640 | [[https://www.fpbase.org/spectra/?s=1201&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|Semrock FF01-440/521/607/700]] | |
| + | | 2 | c405s | 445/45 | 405/488/561/640 | [[https://www.fpbase.org/spectra/?s=814&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|Semrock FF01-445/45]] | | ||
| + | | 3 | c488s | 525/30 | 405/488/561/640 | [[https://www.fpbase.org/spectra/?s=902&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|Semrock FF01-525/30]] | | ||
| + | | 4 | c561s | 617/73 | 405/488/561/640 | [[https://www.fpbase.org/spectra/?s=2874&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|Semrock FF02-617/73]] | | ||
| + | | 5 | c594s | 642/80 | 445/515/594 | [[https://www.fpbase.org/spectra/?s=716&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|Chroma ET642/80]] | | ||
| + | | 6 | c637s | 692/40 | 405/488/561/640 | [[https://www.fpbase.org/spectra/?s=1053&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|Semrock FF01-692/40]] | | ||
| + | | 7 | Dual (c###d) | 525 642 | 405/488/561/640 | [[https://www.fpbase.org/spectra/?s=5890&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|Semrock EM01-R488/568]] | | ||
| + | | 8 | c445s | 482/25 | 445/515/594 | [[https://www.fpbase.org/spectra/?s=861&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|Semrock FF01-482/25]] | | ||
| + | | 9 | c488/YFP | 542/27 | 405/488/561/640 | [[https://www.fpbase.org/spectra/?s=716&showY=0&showX=1&showGrid=0&areaFill=1&logScale=0&scaleEC=0&scaleQY=0&shareTooltip=1&palette=wavelength|Semrock FF01-542/27]] | | ||
| + | | 10 | cBF-DIC | - | any | - | | ||
| ==== Nikon Ti2 filterset cubes ==== | ==== Nikon Ti2 filterset cubes ==== | ||
| - | ^ Position ^ Filterset ^ Excitation ^ Dichroic ^ Emission ^ Reference ^ | + | ^ Turret Position ^ Filter cube ^ Excitation ^ Dichroic ^ Emission ^ |
| - | | | | | | | | | + | | 4 (lower deck) | DAPI | 378/52 | 409 | BP 447/60 | |
| + | | 2 (lower deck) | GFP | 472/30 | 495 | BP 520/35 | | ||
| + | | 3 (lower deck) | Texas Red | 562/40 | 593 | BP 624/40 | | ||
| + | | 6 (lower deck) | CSU (Confocal) | - | empty | - | | ||
| + | | 3 (upper deck) | Phasor/Ablate | - | <380 | - | | ||
| ==== Objectives ==== | ==== Objectives ==== | ||
| ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ||
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| + | **Upon request:** | ||
| + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ||
| + | | 40x | Apochromat | Water | 1.25 | 0.2–0.16 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-1923|MRD77400]] | | ||
| ==== Camera ==== | ==== Camera ==== | ||
| + | |||
| + | ^ Model ^ Frame Size ^ Pixel Size (µm) ^ Quantum Efficiency ^ | ||
| + | | [[https://www.teledynevisionsolutions.com/products/prime-95b/?segment=tvs&vertical=tvs%20-%20photometrics|Photometrics Prime 95B]] | 1200 x 1200 | 11 x 11 | > 95% | | ||
| + | |||
| + | |||
| + | ===== Microscope Turn On Procedure ===== | ||
| + | |||
| + | * Turn on the master power bar (left side of the table) | ||
| + | {{Oeiras_Marianas_TurningOn_1.jpg?|}} | ||
| + | * Turn the laser key to the On position | ||
| + | {{Oeiras_Marianas_TurningOn_2.jpg?|}} | ||
| + | * If your using temperature and/or CO2, turn on OkoLab power bar (left ide of the table) | ||
| + | {{Oeiras_Marianas_TurningOn_3_b.jpg?|}} | ||
| + | * After 2 minutes, turn on PC (left side of the table) | ||
| + | {{Oeiras_Marianas_TurningOn_4.jpg?|}} | ||
| + | * Login in to Windows with your Agendo credentials | ||
| + | * Wait 2 min to allow for system components detection | ||
| + | * Start the **SlideBook** software | ||
| + | |||
| + | ===== Microscope Turn Off Procedure ===== | ||
| + | |||
| + | If there is another user for this microscope during the day: | ||
| + | * Exit the **SlideBook** software, leave the microscope on | ||
| + | * Log off the computer | ||
| + | * Clean up immersion objectives | ||
| + | |||
| + | Else: | ||
| + | * Exit the **SlideBook** software | ||
| + | * Shut down the computer | ||
| + | * Turn off incubation/CO2 on the powerbar (if used) | ||
| + | * Switch laser key off | ||
| + | * Turn off the master power bar | ||
| ---- | ---- | ||
| [[resources|Back to the Equipment page]] | [[resources|Back to the Equipment page]] | ||
oeiras_3i_marianas_sdc.1736262554.txt.gz · Last modified: 2025/01/07 16:09 by bioimaging
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