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nikon_htm-hcs [2025/01/09 11:16] bioimaging [Objectives] |
nikon_htm-hcs [2025/03/28 17:07] (current) bioimaging [Microscope overview] |
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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). | ||
- | ===== System components ===== | + | |
+ | ===== LED 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 ===== | ||
^ Position ^ Filterset ^ Reference ^ Excitation ^ Dichroic ^ Emission ^ | ^ 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 ===== | ||
+ | |||
+ | ^ 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) ^ | ||
+ | | 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 | | ||
+ | | 20x | Plan Apo Lambda | Air | 0.75 | 1 | Yes | Yes (N1) | No | MRD00205* | 0.323 | 0.216 | | ||
+ | | 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 | ||
+ | |||
+ | The Bioimaging Platform **highly advises against using the 40x LWD objective with 1.5x zoom** | ||
+ | ===== Camera ===== | ||
+ | |||
+ | ^ 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% | | ||
+ | ===== Microscope Turn On Procedure ===== | ||
- | ==== Objectives ==== | + | * Turn on the CoolLed controller on **top of the microscope's shelf**. |
- | ^ Position ^ Magnification ^ Model ^ Immersion ^ NA ^ WD (mm) ^ PFS compatible ^ DIC polarizer ^ Reference ^ | + | {{nikon-htm_turnon_1.png?200|}} |
- | | 1 | 10x | Plan Apo | Air | 0.45 | 4 | No | N1 | [[https://www.microscope.healthcare.nikon.com/en_EU/products/optics/selector/comparison/-179798|MRD70170]] | | + | * Turn on the microscope using the **switch** on the **bottom of the microscope's optical table**. |
- | | 2 | 20x | Plan Apo Lambda | Air | 0.75 | 1 | Yes | N1 | MRD00205* | | + | {{nikon-htm_turnon_2.png?200|}} |
- | | 3 | 100x | Plan Apo Lambda | Oil | 1.45 | 0.13 | Yes | N2 | MRD01905* | | + | |
- | | 4 | 40x | Plan Fluor | Oil | 1.3 | 0.24 | Yes | N2 | [[https://www.microscope.healthcare.nikon.com/en_EU/products/optics/selector/comparison/-1829|MRH01401]] | | + | |
- | | 5 | 40x | Plan Apo | Air | 0.95 | 0.17-0.25 | Yes | N2 | MRD00400* | | + | |
- | | 6 | 40x | LWD* * | Air | 0.55 | 2.1 | No | NA* * * | [[https://www.microscope.healthcare.nikon.com/en_EU/products/optics/selector/comparison/-1683|MRP46402]] | | + | |
- | * Not fabricated anymore ; * * LWD - Long Working Distance ; * * * NA - Not applicable | ||
- | ==== Camera ==== | ||
+ | ===== Microscope Turn Off Procedure ===== | ||
+ | * Turn off the microscope using the **switch** on the **bottom of the microscope's optical table**. | ||
+ | {{nikon-htm_turnon_2.png?200|}} | ||
+ | * Turn off the CoolLed controller on **top of the microscope's shelf**. | ||
+ | {{nikon-htm_turnon_1.png?200|}} | ||
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[[resources|Back to the Equipment page]] | [[resources|Back to the Equipment page]] |