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Nikon HCS (High Content Screening) | Reservations

  • NikonHCS

  • 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:

Suggestion for description in "Materials and Methods":

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.

Suggestion for "Acknowledgments":

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).

Maintenance log:

  • 03-03-2015 - Izasa technician finalized setup after changed to the System. Macro, camera angle, etc.
  • 19-02-2015 - Oko Labs incubator installed.
  • 20_21-01-2015 - Microscope working and training sessions taking place to teach new users how to use the microscope.

Booking Calendar details

Configuration
Value

Available objectives

Place mouse over Magnification to see the objective.
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
PLAN APO λ
0.45
-
4
0.712
0.650
0.433
1.068
20x
PLAN APO λ
0.75
-
1
0.350
0.325
0.217
0.641
40x
Plan Fluor
1.3
Oil
0.24
0.202
0.162
0.108
0.370
100x
PLAN APO λ
1.45
Oil
0.13
0.181
0.065
0.043
0.331
20x
PLAN Fluor
0.75
MImm
0.51-0.33
0.350
0.325
0.217
0.641
60x
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

Installed filtersets

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

Spectra, lines & filters:

Setup

Turn on procedures

  1. Turn on the button 1.

    Stage Power

  2. Turn on the lamp power source (LEDs).

    Lamp Power

  3. Turn on the power bar on the right side of the table.

    Power bar

  4. Start the Nikon Elements software available at the Desktop.

    NIS Elements shortcut

  5. Log in to the software using the USERS account.

    NIS Elements Log-in

  6. Set up your experiments on the microscope.

Turn off procedures

  1. Transfer your data. If you need remote access to the PC, ask the facility staff.
  2. Clean all immersion lenses you used.
  3. Close the software and log off the computer.
  4. If the next user is scheduled in more than 1 hour, turn off everything:
  5. Turn off the button 1.
  6. Turn off the LED lamp (button 2).
  7. Turn off the power bar (button 3).

Acquisition

  • Grid Colection / Tile

    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.

  • Tile Scan

  • Perfect Focus System (PFS)

    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.

  • Nikon PFS

Extra Info