Bioimaging Wiki

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<b>Location</b>: [Oeiras] Room 0B02<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>Nickname</b>: Nikon HTM-HCS<br>
<b>Software</b>: NIS-Elements <br>
<b>Year</b>: 2015 <br>
<b>SN</b>:  <br>
&nbsp <br>
Data will be deleted after: <b>1 month</b>
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&rarr; <a href="/facility/bioimaging/doku.php?id=nikon_htm-hcs_quality"><img src="/facility/bioimaging/lib/exe/fetch.php?media=list-icon.png" width=18> Nikon HTM-HCS Quality Control</a>
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&rarr; <a href="/facility/bioimaging/doku.php?id=nikon_htm-hcs_usage"><img src="/facility/bioimaging/lib/exe/fetch.php?media=chart_line.png">  Nikon HTM-HCS Usage Statistics</a>
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===== Microscope overview =====

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 optics available include fluorescence filters and brightfield, including DIC (for several magnifications) and phase-contrast (40x LWD only).


===== 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  ^
|  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 =====

  * 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**.
{{nikon-htm_turnon_2.png?200|}}


===== 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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