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oeiras_3i_marianas_sdc_bsl2 [2025/01/08 15:58] bioimaging [Camera] |
oeiras_3i_marianas_sdc_bsl2 [2026/09/08 01:19] (current) bioimaging |
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| - | <img src="/facility/bioimaging/lib/exe/fetch.php?media=oeiras_3i_marianas_sdc_ii_tirf_name_gimm_2024.png"> | + | <img src="/facility/bioimaging/lib/exe/fetch.php?media=oeiras_3i_marianas_sdc_sora_tirf_bsl2_name.png"> |
| <table style="border:0px solid white;"> | <table style="border:0px solid white;"> | ||
| <tr style="border:0px solid white;"> | <tr style="border:0px solid white;"> | ||
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| <b>Nickname</b>: "Marianas BSL2"<br> | <b>Nickname</b>: "Marianas BSL2"<br> | ||
| <b>Software</b>: SlideBook 2024<br> | <b>Software</b>: SlideBook 2024<br> | ||
| - | <b>Year</b>: 2022 <br> | + | <b>Year</b>: 2022 <br> |
| - | <b>SN</b>: <br> | + | <b>SN</b>: CS-02136<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 ===== | ||
| - | {{ :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. | + | {{ :yokogawa_sd.png?350|}} The 3i Marianas SDC (SoRa + TIRF) 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 SoRa 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 Super-resolution by Optical Re-Assignment (SoRa) scanhead allows 1x, 2.8x and 4x magnification (for extra magnification and to allow Nyquist sampling and deconvolution). SoRa images have a 1.4x resolution improvement and deconvolved SoRa images have a 2x resolution improvement compared to standard spinning disk data. |
| + | The system also allows for Total Internal Reflection Fluorescence (TIRF) imaging, which uses an evanescent wave with very low penetration depth, perfect for imaging live cell focal adhesions, membrane dynamics, and receptor function as well as in vitro experiments that require the thinnest optical section. | ||
| * **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]] | ||
| * **Confocal scanner**: [[https://www.yokogawa.com/eu/solutions/products-and-services/life-science/spinning-disk-confocal/csu-w1-confocal-scanner-unit/|Yokogawa CSU-W1]] + [[https://www.yokogawa.com/solutions/products-and-services/life-science/spinning-disk-confocal/uniformizer/#Overview|Uniformizer]] | * **Confocal scanner**: [[https://www.yokogawa.com/eu/solutions/products-and-services/life-science/spinning-disk-confocal/csu-w1-confocal-scanner-unit/|Yokogawa CSU-W1]] + [[https://www.yokogawa.com/solutions/products-and-services/life-science/spinning-disk-confocal/uniformizer/#Overview|Uniformizer]] | ||
| * **Camera**: [[https://www.teledynevisionsolutions.com/en-150/products/prime-95b/?segment=tvs&vertical=tvs%20-%20photometrics|Photometrics Prime 95B]] | * **Camera**: [[https://www.teledynevisionsolutions.com/en-150/products/prime-95b/?segment=tvs&vertical=tvs%20-%20photometrics|Photometrics Prime 95B]] | ||
| + | * **Features** | ||
| + | * | Super-Resolution by Optical Re-Assignment (SoRa) | ||
| + | * | Phasor Holographic Photomanipulation | ||
| + | * | Vector2 TIRF | ||
| {{:warning.png?25|}} Data files older than **1 month** will be automatically deleted on this system, please copy your data to the GIMM server using the desktop link. | {{:warning.png?25|}} Data files older than **1 month** will be automatically deleted on this system, please copy your data to the GIMM server using the desktop link. | ||
| ===== System components ===== | ===== System components ===== | ||
| - | ==== Lasers ==== | + | ==== 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 ==== | + | ^ LASER ^ Wavelength ^ Maximum Power ^ |
| + | | Violet | <fc #a600ff>405 nm</fc> | 100 mW | | ||
| + | | Blue-Violet | <fc #0028ff>445 nm</fc> | 150 mW | | ||
| + | | Blue | <fc #00f7ff>488 nm</fc> | 150 mW | | ||
| + | | Yellow-Green | <fc #c6ff00>561 nm</fc> | 100 mW | | ||
| + | | Orange | <fc #ffd200>594 nm</fc> | 100 mW | | ||
| + | | Red | <fc #ff3000>637 nm</fc> | 140 mW | | ||
| - | ^ Position ^ Filter name ^ Transmission (nm) ^ Dichroic ^ Reference ^ | + | ==== Filtersets (eyepieces) ==== |
| - | | 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 | - | | + | |
| + | ^ Position ^ Filterset ^ Excitation ^ Dichroic ^ Emission ^ | ||
| + | | 4 (lower deck) | Blue | 352-404 nm | 409 | <fc #0035ff>417-477 nm</fc> | | ||
| + | | 2 (lower deck) | Green | 457-487 nm | 495 nm | <fc #36ff00>503-538 nm</fc> | | ||
| + | | 3 (lower deck) | Orange | 542-582 nm | 593 nm | <fc #ff6700>604-644 nm</fc> | | ||
| + | | 6 (lower deck) | CSU (Confocal) | - | - | - | | ||
| ==== Objectives ==== | ==== Objectives ==== | ||
| + | |||
| ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Reference ^ | ||
| - | | | | | | | | + | | <fc #fef042>10x</fc> | CFI Plan Apochromat | Air | 0.45 | 4 |[[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-179798 |MRD70170]] | |
| + | | <fc #0aa05c>20x</fc> | CFI Plan Fluor | Oil, W, G, Sil | 0.75 | 0.51–0.33 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-1827 |MRH07241]] | | ||
| + | | <fc #01b3f5>40x</fc> | CFI Plan Apochromat | Air | 0.95 | 0.21 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-179806 |MRD70470]] | | ||
| + | | <fc #2c80c0>60x</fc> | CFI Plan Apochromat | Water | 1.20 | 0.31–0.28 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-1855|MRD07602]] | | ||
| + | | <fc #2c80c0>60x</fc> | CFI Plan Apochromat | Oil | 1.42 | 0.15 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-179808|MRD71670]] | | ||
| + | | 100x | CFI Apochromat | Oil | 1.49 | 0.16–0.09 | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-1932|MRD01991]] | | ||
| + | |||
| + | ==== Emission Filters CSU W1 ==== | ||
| + | |||
| + | ^ Position ^ Filter name ^ Dichroic ^ Transmission ^ Reference ^ | ||
| + | | 1 | QUAD (c###q) | 405/488/561/640 | <fc #0000ff>420-460 nm</fc> | [[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]] | | ||
| + | | ::: | ::: | ::: | <fc #3aff00>511-532 nm</fc> | ::: | | ||
| + | | ::: | ::: | ::: | <fc #ffa600>590-624 nm</fc> | ::: | | ||
| + | | ::: | ::: | ::: | <fc #ff0000>678-723 nm</fc> | ::: | | ||
| + | | 2 | c405s | 405/488/561/640 | <fc #0028ff>423-468 nm</fc> | [[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 | 405/488/561/640 | <fc #4aff00>510-540 nm</fc> | [[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 | 405/488/561/640 | <fc #ff8200>581-654 nm</fc> | [[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 | 445/515/594 | <fc #ff2100>602-682 nm</fc> | [[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 | 405/488/561/640 | <fc #ff0000>672-712 nm</fc> | [[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) | 405/488/561/640 | <fc #4aff00>510-540 nm</fc> | [[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]] | | ||
| + | | ::: | ::: | ::: | <fc #ff2100>602-682 nm</fc> | ::: | | ||
| + | | 8 | c445s | 445/515/594 | <fc #00deff>470-495 nm</fc> | [[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 | 405/488/561/640 | <fc #88ff00>529-556 nm</fc> | [[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 | - | | ||
| ==== Camera ==== | ==== Camera ==== | ||
oeiras_3i_marianas_sdc_bsl2.1736348280.txt.gz · Last modified: 2025/01/08 15:58 by bioimaging
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