oeiras_3i_marianas_sdc_bsl2
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oeiras_3i_marianas_sdc_bsl2 [2025/01/08 15:35] bioimaging [Microscope overview] |
oeiras_3i_marianas_sdc_bsl2 [2025/01/08 16:08] (current) bioimaging [Objectives] |
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| ===== Microscope overview ===== | ===== Microscope overview ===== | ||
| - | {{ :yokogawa_sd.png?350|}} The 3i Marianas SDC II //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 II super-resolution //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. |
| - | 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, TIRF, a removable heating stage with CO2 delivery, and a photomanipulation module (Phasor). The scanhead allows 1x, 2.8x and 4x 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, TIRF, a removable heating stage with CO2 delivery, and a photomanipulation module (Phasor). The scanhead SoRa allows 1x, 2.8x and 4x magnification (for extra magnification and to allow Nyquist sampling and deconvolution). |
| * **Microscope**: [[https://www.microscope.healthcare.nikon.com/products/inverted-microscopes/eclipse-ti2-series/|Nikon Ti2 Eclipse]] | * **Microscope**: [[https://www.microscope.healthcare.nikon.com/products/inverted-microscopes/eclipse-ti2-series/|Nikon Ti2 Eclipse]] | ||
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| ===== System components ===== | ===== System components ===== | ||
| - | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ | + | ==== Lasers ==== |
| - | | | | | | | | | + | ^ Laser line ^ Wavelength ^ Maximum Power ^ |
| + | | 405 | 405 nm | 100 mW | | ||
| + | | 445 | 445 nm | 150 mW | | ||
| + | | 488 | 488 nm | 150 mW | | ||
| + | | 561 | 561 nm | 100 mW | | ||
| + | | 594 | 594 nm | 100 mW | | ||
| + | | 637 | 637 nm | 140 mW | | ||
| + | |||
| + | ==== Emission Filters CSU W1 unit ==== | ||
| + | |||
| + | ^ 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 | not config | 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 ==== | ||
| + | |||
| + | ^ 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 | - | | ||
| ==== Objectives ==== | ==== Objectives ==== | ||
| ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ||
| - | | | | | | | | + | | 10x | Plan Apochromat | Air | 0.45 | 4 |[[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-179798 |MRD70170]] | |
| + | | 20x | Plan Fluor | Oil, W, G, Sil | 0.75 | 0.51–0.33 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-1827 |MRH07241]] | | ||
| + | | 40x | Plan Apochromat | Air | 0.95 | 0.21 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-179806 |MRD70470]] | | ||
| + | | 60x | Plan-Apochromat | Water | 1.20 | 0.31–0.28 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-1855|MRD07602]] | | ||
| + | | 60x | Plan Apochromat | Oil | 1.42 | 0.15 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-179808|MRD71670]] | | ||
| + | | 100x | Apochromat TIRF | Oil | 1.49 | 0.16–0.09 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-1932|MRD01991]] | | ||
| ==== 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% | | ||
| ---- | ---- | ||
| [[resources|Back to the Equipment page]] | [[resources|Back to the Equipment page]] | ||
oeiras_3i_marianas_sdc_bsl2.1736346942.txt.gz · Last modified: 2025/01/08 15:35 by bioimaging
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