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The Nikon HCS microscope is a fully automated microscope specially designed to for high throughput and screening of samples with the help of fully programable and customizable acquisition protocols. Equipped with the CoolLED light source with 4 excitation bands. The camera is a 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).
Location :
Microscope : Nikon Ti
Year Installed: 2015
Optics : Fluorescence and brightfield (with DIC)
Camera : Andor Zyla 4.2 sCMOS 4.2Mpx
Operating conditions : Temperature and CO2 control
Keywords:
Images were acquired on a comercial Nikon High Content Screening microscope, based on Nikon Ti equiped with a Andor Zyla 4.2 sCMOS camera, using the a 100x 1.45 NA objective, DAPI and FITC fluorescence filtersets and controlled with the Nikon Elements software.
We would like to thank the technical support of IGC's Advanced Imaging Facility (AIF-UIC), which is supported by the national Portuguese funding ref# PPBI-POCI-01-0145-FEDER-022122, co-financed by Lisboa Regional Operational Programme (Lisboa 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (FEDER) and Fundação para a Ciência e a Tecnologia (FCT; Portugal).
Configuration |
Value |
Magnification |
Objective type |
NA1 |
Immersion |
Working distance (mm) |
Resolution2 @525nm in XY (µm) |
Pixel Size (µm) |
Pixel Size * 1.5x (µm) |
Nyquist Sampling @525nm in Z (µm) |
Link |
|
10x |
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PLAN APO λ |
0.45 |
- |
4 |
0.712 |
0.650 |
0.433 |
1.068 |
|
20x |
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PLAN APO λ |
0.75 |
- |
1 |
0.350 |
0.325 |
0.217 |
0.641 |
|
40x |
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Plan Fluor |
1.3 |
Oil |
0.24 |
0.202 |
0.162 |
0.108 |
0.370 |
|
100x |
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PLAN APO λ |
1.45 |
Oil |
0.13 |
0.181 |
0.065 |
0.043 |
0.331 |
|
20x |
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PLAN Fluor |
0.75 |
MImm |
0.51-0.33 |
0.350 |
0.325 |
0.217 |
0.641 |
|
60x |
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PLAN APO |
1.27 |
water |
0.18-0.16 |
0.207 |
0.119 |
0.079 |
0.378 |
(1) NA - Numerical Aperture. For more information, follow this link.
(2) These values were calculated using the Abbe resolution formula: R = 0.5*λ/NA
Filter |
Excitation band |
Dichroic |
Emission band |
Filter cube name |
Filter position |
Quad DAPI |
BP 378/58 |
409/493/573/652 |
BP 432/40 |
QUAD |
1 |
Quad GFP |
BP 474/30 |
BP 515/36 |
|||
Quad TRITC |
BP 554/26 |
BP 595/36 |
|||
Quad Cy5 |
BP 635/22 |
BP LP 658 |
|||
FITC |
BP 474/27 |
495 |
BP 525/45 |
FITC |
2 |
TRITC |
BP 543/22 |
562 |
BP 593/40 |
TRITC |
3 |
Cy5 |
BP 635/18 |
652 |
BP 680/42 |
Cy5 |
4 |
Fast Filter Wheel sets. These are limited to the Quad emission bands.
Filter name on NIS |
Emission band |
Link |
QUAD-380nm |
BP 460/50 |
|
QUAD-470 nm FFW |
BP 525/50 |
|
QUAD-550 nm FFW |
BP 595/50 |
|
QUAD-635 nm FFW |
BP 685/40 |
It is possible to acquire grids and tiles of big samples through NIS Elements. For a more detailed protocol, please check the following link or click in the image to the right.
Nikon has a hardware autofocus system called Perfect Focus System (PFS) which allows continuous sample focusing using the coverslip position as the reference. For more detailed information, follow the link or click the image on the right.