nikon_htm-hcs
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nikon_htm-hcs [2025/02/04 15:31] bioimaging [Objectives] |
nikon_htm-hcs [2025/12/12 11:29] (current) bioimaging |
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| <a href="/facility/bioimaging/lib/exe/fetch.php?media=nikon_htm-hcs.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=nikon_htm-hcs.jpg" width=300></a> | <a href="/facility/bioimaging/lib/exe/fetch.php?media=nikon_htm-hcs.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=nikon_htm-hcs.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 0B02<br> | + | <b>Location</b>: <a href="/facility/bioimaging/lib/exe/fetch.php?media=bartolomeu_dias_wing_2025_-_nikon_htm-hcs.png">[Oeiras] Room 0B05</a><br> |
| <b>Manufacturer</b>: <a href="https://www.microscope.healthcare.nikon.com/en_EU/" target="_blank">Nikon</a> <br> | <b>Manufacturer</b>: <a href="https://www.microscope.healthcare.nikon.com/en_EU/" target="_blank">Nikon</a> <br> | ||
| <b>Model</b>: <a href="https://www.microscope.healthcare.nikon.com/en_EU/products/inverted-microscopes/eclipse-ti-series" target="_blank">Nikon Eclipse Ti</a> <br> | <b>Model</b>: <a href="https://www.microscope.healthcare.nikon.com/en_EU/products/inverted-microscopes/eclipse-ti-series" target="_blank">Nikon Eclipse Ti</a> <br> | ||
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| ===== Microscope overview ===== | ===== Microscope overview ===== | ||
| - | The Nikon-HTM/HCS microscope is a fully automated microscope capable of high-throughput imaging and screening of samples (eg, Ibidi chamber slides or well-plates); with the help of fully programmable and customizable acquisition protocols. | + | The Nikon HTM-HCS is a fully automated microscope capable of high-throughput imaging and screening of samples (eg, Ibidi chamber slides or well-plates); with the help of fully programmable and customizable acquisition protocols. |
| The system is equipped with a CoolLED fluorescence illuminator with 4 excitation bands, and an Andor Zyla 4.2 sCMOS camera with 4.0 MPixels with a large field of view capable of maximum of 100 FPS at full frame (2048x2048). | The system is equipped with a CoolLED fluorescence illuminator with 4 excitation bands, and an Andor Zyla 4.2 sCMOS camera with 4.0 MPixels with a large field of view capable of maximum of 100 FPS at full frame (2048x2048). | ||
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| The optics available include fluorescence filters and brightfield, including DIC (for several magnifications) and phase-contrast (40x LWD only). | The optics available include fluorescence filters and brightfield, including DIC (for several magnifications) and phase-contrast (40x LWD only). | ||
| + | {{::ionoptix_realtime-acquisition-cell-length-600x500.jpg?200 |}} | ||
| + | This microscope is also equipped with a [[https://www.ionoptix.com/products/systems/calcium-and-contractility-system/|Ionoptix Calcium and Contractility system]] for high-speed simultaneous acquisition of Fura-2 fluorescence with video-based cell length measurement to study myocyte function, contracting spontaneously or with field stimulation. | ||
| - | ===== LED lines ===== | + | * **High-speed contractility camera**: [[https://www.ionoptix.com/products/components/sensors/myocam-s3/|MyoCam-S3]] |
| + | * **Electrical Field Stimulator**: [[https://www.ionoptix.com/products/components/stimulators/myopacer-cell-stimulator/|MyoPacer]] | ||
| + | * **LED Light Source**: [[https://www.ionoptix.com/products/components/light-sources/led-light-source/|UV LEDs for Fura-2 excitation]] | ||
| + | * **High-speed PMT**: [[https://www.ionoptix.com/products/components/sensors/pmt-sub-system/|PMT400]] for photon counting at 1kHz | ||
| + | * **Software**: [[https://www.ionoptix.com/products/software/ionwizard-core-and-analysis/|IonWizard]] 7.7.3.180 | ||
| + | |||
| + | ===== System Components ===== | ||
| + | |||
| + | ==== LEDs ==== | ||
| This system is equipped with a CoolLED fluorescence illuminator with 4 excitation lines: | This system is equipped with a CoolLED fluorescence illuminator with 4 excitation lines: | ||
| - | * 380 nm - E.g. DAPI | ||
| - | * 470 nm - E.g. FITC | ||
| - | * 550 nm - E.g. TRITC | ||
| - | * 635 nm - E.g. Cy5 | ||
| - | ===== Filterset cubes ===== | + | ^ Line ^ Wavelength ^ |
| + | | UV | <fc #be00cc>380 nm</fc> | | ||
| + | | B | <fc #00a9ff>470 nm</fc> | | ||
| + | | G | <fc #a3ff00>550 nm</fc> | | ||
| + | | R | <fc #ff3900>635 nm</fc> | | ||
| - | ^ Position ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ | ||
| - | | 1 | Quad-DAPI | | 378/58 | [[https://www.idex-hs.com/store/product-detail/ff409_493_573_652_di02_25x36/fl-007110?search=true|409]] | 432/40 | | ||
| - | | 1 | Quad-FITC | | 474/30 | [[https://www.idex-hs.com/store/product-detail/ff409_493_573_652_di02_25x36/fl-007110?search=true|493]] | 515/36 | | ||
| - | | 1 | Quad-TRITC | | 554/26 | [[https://www.idex-hs.com/store/product-detail/ff409_493_573_652_di02_25x36/fl-007110?search=true|573]] | 595/36 | | ||
| - | | 1 | Quad-Cy5 | | 635/22 | [[https://www.idex-hs.com/store/product-detail/ff409_493_573_652_di02_25x36/fl-007110?search=true|652]] | LP 658 | | ||
| - | | 2 | FITC | | 474/27 | 495 | 525/45 | | ||
| - | | 3 | TRITC | | 543/22 | 562 | 593/40 | | ||
| - | | 4 | Cy5 | | 635/18 | 652 | 680/42 | | ||
| - | ===== Fast Filter Wheel emission filters ===== | + | ==== Filtersets Turret 1 (top) ==== |
| + | |||
| + | ^ Position ^ Filterset ^ Excitation ^ Dichroic ^ Emission ^ Reference ^ | ||
| + | | 1 | QUAD | 349-407 nm | 409 nm | <fc #3300ff>412-452 nm</fc> | [[https://www.idex-hs.com/store/product-detail/ff409_493_573_652_di02_25x36/fl-007110?search=true|FL-007110]] | | ||
| + | | ::: | ::: | 459-489 nm | 493 nm | <fc #1fff00>497-533 nm</fc> | ::: | | ||
| + | | ::: | ::: | 541-567 nm | 573 nm | <fc #ffcf00>577-613 nm</fc> | ::: | | ||
| + | | ::: | ::: | 624-646 nm | 652 nm | <fc #ff0000>> 658 nm</fc> | ::: | | ||
| + | | 2 | Green | 460-488 nm | 495 nm | <fc #4aff00>503-548 nm</fc> | | | ||
| + | | 3 | Orange | 532-554 nm | 562 nm | <fc #ffd500>573-613 nm</fc> | | | ||
| + | | 4 | Red | 624-642 nm | 652 nm | <fc #ff0000>659-701 nm</fc> | | | ||
| + | |||
| + | |||
| + | ==== Filtersets Turret 2 (bottom) ==== | ||
| + | |||
| + | ^ Position ^ Filterset ^ Excitation ^ Dichroic ^ Emission ^ | ||
| + | | 6 | Fura-2 | 334-346 nm | 400 nm | <fc #00ff28>498-518 nm</fc> | | ||
| + | | ::: | ::: | 370-390 nm | ::: | ::: | | ||
| + | |||
| + | ==== Emission Filters Fast Filter Wheel ==== | ||
| + | |||
| + | ^ Filterset ^ Emission ^ Reference ^ | ||
| + | | DAPI-FW | <fc #007bff>435-485 nm</fc> | [[https://www.chroma.com/products/parts/at460-50m|AT460/50m]] | | ||
| + | | FITC-FW | <fc #4aff00>500-550 nm</fc> | [[https://www.chroma.com/products/parts/et525-50m|ET525/50m]] | | ||
| + | | TRITC-FW | <fc #ffcf00>570-620 nm</fc> | [[https://www.chroma.com/products/parts/et595-50m|ET595/50m]] | | ||
| + | | Cy5-FW | <fc #ff0000>665-705 nm</fc> | [[https://www.chroma.com/products/parts/s685-40m|S685/40m]] | | ||
| + | |||
| + | ==== Objectives ==== | ||
| - | ^ Filterset ^ Reference ^ Transmission ^ | ||
| - | | DAPI-FW | [[https://www.chroma.com/products/parts/at460-50m|at460-50m]] | 460/50 | | ||
| - | | FITC-FW | [[https://www.chroma.com/products/parts/et525-50m|et525-50m]] | 525/50 | | ||
| - | | TRITC-FW | [[https://www.chroma.com/products/parts/et595-50m|et595-50m]] | 595/50 | | ||
| - | | Cy5-FW | [[https://www.chroma.com/products/parts/s685-40m|s685-40m]] | 685/40 | | ||
| - | ===== Objectives ===== | ||
| ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ PFS ^ DIC ^ PhC ^ Reference ^ Pixel size (um) (1x zoom) ^ Pixel size (um) (1.5x zoom) ^ | ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ PFS ^ DIC ^ PhC ^ Reference ^ Pixel size (um) (1x zoom) ^ Pixel size (um) (1.5x zoom) ^ | ||
| - | | 10x | Plan Apo | Air | 0.45 | 4 | No | Yes (N1) | No | [[https://www.microscope.healthcare.nikon.com/en_EU/products/optics/selector/comparison/-179798|MRD70170]] | 0.643 | 0.429 | | + | | <fc #fef042>10x</fc> | Plan Apo | Air | 0.45 | 4 | No | Yes (N1) | No | [[https://www.microscope.healthcare.nikon.com/en_EU/products/optics/selector/comparison/-179798|MRD70170]] | 0.643 | 0.429 | |
| - | | 20x | Plan Apo Lambda | Air | 0.75 | 1 | Yes | Yes (N1) | No | MRD00205* | 0.323 | 0.216 | | + | | <fc #0aa05c>20x</fc> | Plan Apo Lambda | Air | 0.75 | 1 | Yes | Yes (N1) | No | MRD00205* | 0.323 | 0.216 | |
| + | | <fc #01b3f5>40x</fc> | Plan Fluor | Oil | 1.3 | 0.24 | Yes | Yes (N2) | No | [[https://www.microscope.healthcare.nikon.com/en_EU/products/optics/selector/comparison/-1829|MRH01401]] | 0.162 | 0.108 | | ||
| + | | <fc #01b3f5>40x</fc> | Plan Apo | Air | 0.95 | 0.17-0.25 | Yes | Yes (N2) | No | MRD00400* | 0.161 | 0.107 | | ||
| + | | <fc #01b3f5>40x</fc> | LWD* * | Air | 0.55 | 2.1 | No | No | Yes | [[https://www.microscope.healthcare.nikon.com/en_EU/products/optics/selector/comparison/-1683|MRP46402]] | 0.161 | 0.108 | | ||
| | 100x | Plan Apo Lambda | Oil | 1.45 | 0.13 | Yes | Yes (N2) | No | MRD01905* | 0.065 | 0.043 | | | 100x | Plan Apo Lambda | Oil | 1.45 | 0.13 | Yes | Yes (N2) | No | MRD01905* | 0.065 | 0.043 | | ||
| - | | 40x | Plan Fluor | Oil | 1.3 | 0.24 | Yes | Yes (N2) | No | [[https://www.microscope.healthcare.nikon.com/en_EU/products/optics/selector/comparison/-1829|MRH01401]] | 0.162 | 0.108 | | ||
| - | | 40x | Plan Apo | Air | 0.95 | 0.17-0.25 | Yes | Yes (N2) | No | MRD00400* | 0.161 | 0.107 | | ||
| - | | 40x | LWD* * | Air | 0.55 | 2.1 | No | No | Yes | [[https://www.microscope.healthcare.nikon.com/en_EU/products/optics/selector/comparison/-1683|MRP46402]] | 0.161 | 0.108 | | ||
| * Not fabricated anymore ; * * LWD - Long Working Distance | * Not fabricated anymore ; * * LWD - Long Working Distance | ||
| The Bioimaging Platform **highly advises against using the 40x LWD objective with 1.5x zoom** | The Bioimaging Platform **highly advises against using the 40x LWD objective with 1.5x zoom** | ||
| - | ===== Camera ===== | + | |
| + | ==== Camera ==== | ||
| ^ Model ^ Frame Size ^ Pixel Size (µm) ^ Quantum Efficiency ^ | ^ Model ^ Frame Size ^ Pixel Size (µm) ^ Quantum Efficiency ^ | ||
| | [[https://andor.oxinst.cn/assets/uploads/products/andor/documents/Zyla-4.2-Specifications-Life-Science.pdf|Andor Zyla 4.2 PLUS]] | 2048 × 2048 | 6.5 x 6.5 | 82% | | | [[https://andor.oxinst.cn/assets/uploads/products/andor/documents/Zyla-4.2-Specifications-Life-Science.pdf|Andor Zyla 4.2 PLUS]] | 2048 × 2048 | 6.5 x 6.5 | 82% | | ||
| - | |||
| ===== Microscope Turn On Procedure ===== | ===== Microscope Turn On Procedure ===== | ||
| - | * Turn on the CoolLed controller on **top of the microscope's shelf**. | ||
| - | {{nikon-htm_turnon_1.png?200|}} | ||
| * Turn on the microscope using the **switch** on the **bottom of the microscope's optical table**. | * Turn on the microscope using the **switch** on the **bottom of the microscope's optical table**. | ||
| - | {{nikon-htm_turnon_2.png?200|}} | + | {{nikon-htm_turnon_3.png?200|}} |
| ===== Microscope Turn Off Procedure ===== | ===== Microscope Turn Off Procedure ===== | ||
| * Turn off the microscope using the **switch** on the **bottom of the microscope's optical table**. | * Turn off the microscope using the **switch** on the **bottom of the microscope's optical table**. | ||
| - | {{nikon-htm_turnon_2.png?200|}} | + | {{nikon-htm_turnon_3.png?200|}} |
| - | * Turn off the CoolLed controller on **top of the microscope's shelf**. | + | |
| - | {{nikon-htm_turnon_1.png?200|}} | + | |
| ---- | ---- | ||
| [[resources|Back to the Equipment page]] | [[resources|Back to the Equipment page]] | ||
nikon_htm-hcs.1738679510.txt.gz · Last modified: 2025/02/04 15:31 by bioimaging
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