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dragonfly [2025/07/19 21:39] bioimaging |
dragonfly [2026/01/20 16:09] (current) bioimaging |
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| <a href="/facility/bioimaging/lib/exe/fetch.php?media=dragonfly.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=dragonfly.jpg" width=300></a> | <a href="/facility/bioimaging/lib/exe/fetch.php?media=dragonfly.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=dragonfly.jpg" width=300></a> | ||
| <td style="border:0px solid white;"> <p style="line-height:1.8"> | <td style="border:0px solid white;"> <p style="line-height:1.8"> | ||
| - | <b>Location</b>: [Oeiras] Room 0B05<br> | + | <b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=bartolomeu_dias_wing_2025_-_dragonfly.png">[Oeiras] Room 0B05</a><br> |
| <b>Manufacturer</b>: <a href="https://andor.oxinst.com/" target="_blank">Oxford Institutes - Andor</a> <br> | <b>Manufacturer</b>: <a href="https://andor.oxinst.com/" target="_blank">Oxford Institutes - Andor</a> <br> | ||
| <b>Model</b>: <a href="https://andor.oxinst.com/products/dragonfly-confocal-microscope-system" target="_blank">Andor Dragonfly 200</a> <br> | <b>Model</b>: <a href="https://andor.oxinst.com/products/dragonfly-confocal-microscope-system" target="_blank">Andor Dragonfly 200</a> <br> | ||
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| ===== Microscope overview ===== | ===== Microscope overview ===== | ||
| - | Spinning disk confocal microscopy (SDC) is a technique well suited for observation of cells and organoids. Compared to conventional widefield microscopy, SDC provides instant optical sectioning, allowing the imaging of cells and shallow tissues (typically a maximum of a few tens of micrometres deep) in 3D. | + | {{ :yokogawa_sd.png?350|}} Spinning disk confocal microscopy (SDC) is a technique well suited for observation of cells and organoids. Compared to conventional widefield microscopy, SDC provides instant optical sectioning, allowing the imaging of cells and shallow tissues (typically a maximum of a few tens of micrometres deep) in 3D. |
| - | Compared to laser scanning confocal microscopes (such as Leica Stellaris, Zeiss Airyscan), the Dragonfly SDC provides much faster acquisition (up to 400 fps, with limited ROI (2048x128), 40 fps full frame), with lower phototoxicity and photobleaching. | + | Compared to laser scanning confocal microscopes (such as Leica Stellaris, ZEISS Airyscan), the Dragonfly SDC provides much faster acquisition (up to 400 fps, with limited ROI (2048x128), 40 fps full frame), with lower phototoxicity and photobleaching. |
| 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 fluorescence. | 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 fluorescence. | ||
| 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 40um). | 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 40um). | ||
| The scanhead allows 1x or 2x magnification (for extra magnification and to allow Nyquist sampling and deconvolution - see table below). | The scanhead allows 1x or 2x magnification (for extra magnification and to allow Nyquist sampling and deconvolution - see table below). | ||
| - | Camera: [[https://andor.oxinst.com/assets/uploads/products/andor/documents/andor-sona-scmos-specifications.pdf|Andor Sona-4BV11]] | + | * **Microscope**: [[https://www.microscope.healthcare.nikon.com/products/inverted-microscopes/eclipse-ti2-series/|Nikon Ti2 Eclipse]] |
| - | ===== System components ===== | + | * **Confocal scanner**: [[https://andor.oxinst.com/products/dragonfly-confocal-microscope-system|Andor Dragonfly 200]] |
| + | * **Camera**: [[https://andor.oxinst.com/assets/uploads/products/andor/documents/andor-sona-scmos-specifications.pdf|Andor Sona-4BV11]] | ||
| - | ==== Lasers ==== | + | {{: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 ===== | ||
| - | ^ Laser lines ^ Maximum power ^ Measured at objective @100% (mW) ^ | + | ==== LASERs ==== |
| - | | 405 nm | 100 mW | 7 | | + | |
| - | | 488 nm | 100 mW | 17 | | + | |
| - | | 561 nm | 100 mW | 20 | | + | |
| - | | 640 nm | 100 mW | 17 | | + | |
| + | ^ LASER ^ Wavelength ^ Maximum Power ^ Measured after objective @ 100% ^ | ||
| + | | Violet | <fc #a600ff>405 nm</fc> | 100 mW | 7 mW | | ||
| + | | Blue | <fc #00f7ff>488 nm</fc> | 100 mW | 17 mW | | ||
| + | | Yellow-Green | <fc #c6ff00>561 nm</fc> | 100 mW | 20 mW | | ||
| + | | Red | <fc #ff2100>640 nm</fc> | 100 mW | 17 mW | | ||
| - | ==== Emission Filterwheel Spinning Disk ==== | + | ==== Objectives ==== |
| - | + | ||
| - | | Filter “Name” ^ Transmission (nm) ^ Description (use for): ^ CSU dichroic ^ | + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Max FoV (1600x1350px) ^ ideal CSU pinhole ^ Pixel Size (zoom 1x) ^ PFS Compatible ^ Reference ^ |
| - | | Quad (pos0) | 445/521/594/698 | (faster switching) | 405/488/561/640 | | + | | <fc #fef042>10x</fc> | CFI Plan Fluor | Air | 0.30 | 16 | 1.3 mm | 25 μm | 1.0561 μm | Yes | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-1825|MRH00105]] | |
| - | | 405 (pos1) | 445/46 nm | DAPI, Hoechst | 405/488/561/640 | | + | | <fc #0aa05c>20x</fc> | CFI Plan Apochromat | Air | 0.8 | 0.8 | 0.68 mm | 40 μm | 0.5097 μm | Yes | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-179802|MRD70270]] | |
| - | | 488 (pos2) | 521/38 nm | GFP, FITC, Alexa488 | 405/488/561/640 | | + | | <fc #01b3f5>40x</fc> | CFI Plan Apochromat λD | Air | 0.95 | 0.21 | 0.34 mm | 40 μm | 0.2580 μm | Yes | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-179806|MRD70470]] | |
| - | | 561 (pos3) | 594/43 nm | A568 | 405/488/561/640 | | + | | <fc #2c80c0>60x</fc> | CFI Plan Apochromat VC | Water | 1.2 | 0.28-0.31 | 0.23 mm | 40 μm | 0.1719 μm | Yes | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-1855|MRD07602]] | |
| - | | 637 (pos4) | 698/77 nm | Cy5, A633, A647 | 405/488/561/640 | | + | |
| - | | 561 extra (pos8) | 606/64 nm | mCherry | 405/488/561/640 | | + | |
| - | | Pol (pos5) | All (50%) | TR-POLR-DIC-FWL | N/A- | | + | |
| + | **Upon request:** | ||
| + | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ Max FoV (1600x1350px) ^ ideal CSU pinhole ^ Pixel Size (zoom 1x) ^ PFS Compatible ^ Reference ^ | ||
| + | | <fc #0aa05c>20x</fc> | CFI Plan Fluor | 0.75 | multi | 0.33-0.51 | 0.68 mm | 25 μm | 0.5097 μm | No | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-1827|MRH07241]] | | ||
| + | | <fc #01b3f5>40x</fc> | CFI Apochromat LWD λS | 1.15 | Water | 0.59-0.61 | 0.34 mm | 40 μm | 0.2580 μm | Yes | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-5994|MRD77410]] | | ||
| + | | 100x | CFI Plan Apochromat λD | 1.45 | Oil | 0.13 | 0.13 mm | 40 μm | 0.1038 μm | Yes | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-179810|MRD71970]] | | ||
| - | ==== Objectives ==== | + | ==== Emission Filters Spinning Disk ==== |
| - | + | ||
| - | ^ Objectives ^ NA ^ Imm. ^ WD (mm) ^ Max FoV (1600x1350px) ^ ideal CSU pinhole ^ Theor. Resol. um ^ Measured pixel size in μm (1x zoom) ^ PFS Compatible ^ | + | |
| - | | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-1825|10x]] | 0.3 | Dry | 16 | 1.3mm | 25um | 1.07/0.43 | 1.0561 | Yes | | + | |
| - | | [[https://www.microscope.healthcare.nikon.com/products/optics/selector/comparison/-179802|20x]] | 0.8 | Dry | 0.8 | 0.68mm | 40um | 0.40/0.16 | 0.5097 | Yes | | + | |
| - | | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-179806|40x]] | 0.95 | Dry | 0.25-0.17 | 0.34mm | 40um | 0.34/0.13 | 0.2580 | Yes | | + | |
| - | | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-1855|60x]] | 1.2 | Water or Sil | 0.31-0.28 | 0.23mm | 40um | 0.27/0.11 | 0.1719 | Yes | | + | |
| - | | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-1827|20x]]* | 0.75 | Multi | 0.51-0.33 | 0.68mm | 25um | 0.43/0.17 | 0.5097 | No | | + | |
| - | | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-5994|40x]]* | 1.15 | Water/Sil | 0.59-0.61 | 0.34mm | 40um | 0.28/0.11 | 0.2580 | Yes | | + | |
| - | | [[https://www.microscope.healthcare.nikon.com/pt_AMS/products/optics/selector/comparison/-179810|100x]]* | 1.45 | Oil | 0.13 | 0.13mm | 40um | 0.22/0.09 | 0.1038 | Yes | | + | |
| - | *Optional | + | ^ Position ^ Filter name ^ Dichroic ^ Transmission ^ |
| + | | 0 | QUAD | 405/488/561/640 | <fc #0000ff>445 nm</fc> | | ||
| + | | ::: | ::: | ::: | <fc #3aff00>521 nm</fc> | | ||
| + | | ::: | ::: | ::: | <fc #ffd200>594 nm</fc> | | ||
| + | | ::: | ::: | ::: | <fc #ff0000>698 nm</fc> | | ||
| + | | 1 | 405 | 405/488/561/640 | <fc #0028ff>422-468 nm</fc> | | ||
| + | | 2 | 488 | 405/488/561/640 | <fc #3aff00>502-540 nm</fc> | | ||
| + | | 3 | 561 | 405/488/561/640 | <fc #ffd200>573-616 nm</fc> | | ||
| + | | 4 | 637 | 405/488/561/640 | <fc #ff0000>660-724 nm</fc> | | ||
| + | | 5 | Pol | -- | -- | | ||
| + | | 8 | 561 extra | 405/488/561/640 | <fc #4aff00>574-638 nm</fc> | | ||
| ===== Turn On Procedure ===== | ===== Turn On Procedure ===== | ||
dragonfly.1752953994.txt.gz · Last modified: 2025/07/19 21:39 by bioimaging
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