GIMM Bioimaging Wiki

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<a href="/facility/bioimaging/lib/exe/fetch.php?media=oeiras_nikon_htm-hcs.jpg"><img src="/facility/bioimaging/lib/exe/fetch.php?media=oeiras_nikon_htm-hcs.jpg" width=300></a>
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<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>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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&nbsp <br>
&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 [[https://www.biovis.com/images/cameras/SCAS0122E_C11440-42U30.pdf|ORCA-Flash4.0 LT+ sCMOS camera]] with 4.0 MPixels (2048x2048) with a large field of view, low-light sensitivity, and large dynamic range. 

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.

  * **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:

^  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>  |


==== 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 ====

^  Magnification  ^  Model  ^  Immersion  ^  NA  ^  WD (mm)  ^  PFS  ^  DIC  ^  PhC  ^  Reference  ^  Pixel size (um) (1x zoom)  ^  Pixel size (um) (1.5x zoom)  ^
|  <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  |   
|  <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>  |  ELWD* *  |  Air  |  0.60  |  2.80-3.60  |  Yes  |  No  |  Yes  |  [[https://www.microscope.healthcare.nikon.com/en_EU/products/optics/selector/comparison/-1814|MRH48430]]  |  0.161  |  0.108  |   
|  100x  |  Plan Apo Lambda  |  Oil  |  1.45  |  0.13  |  Yes  |  Yes (N2)  |  No  |  MRD01905*  |  0.065  |  0.043  |   

* 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://www.biovis.com/images/cameras/SCAS0122E_C11440-42U30.pdf|ORCA-Flash4.0 LT PLUS]]  |  2048 × 2048  |  6.5 x 6.5  |  82%  |

===== Microscope Turn On Procedure =====

  * Turn on the microscope using the **switch** on the **bottom of the microscope's optical table**.
{{nikon-htm_turnon_3.png?150|}}
  * Turn on the computer 
  * Log In into Windows with your Agendo credentials
  * Start the **NIS** software

===== Microscope Turn Off Procedure =====
If there is another user for this microscope in the next hour:

  * Exit the **NIS** software
  * Log off the computer
  * Clean up immersion objectives  

Else:

  * Exit the **NIS** software
  * Clean up immersion objectives
  * Turn off the computer
  * Turn off the microscope using the **switch** on the **bottom of the microscope's optical table**.
{{nikon-htm_turnon_3.png?150|}}

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