IGC

Nikon HCS (High Content Screening) | Reservations

  • NikonHCS
  • The new Nikon HCS microscope is a fully automated microscopes specially designed to do high troughput and screening of samples by itself for a more efficient image acquisition with the help of fully programable and customizable acquisition protocols. Equiped with PE Fluorescence LED light source as excitation, we are able to do fast imaging of several samples with ease and keeping high levels of fluorescence. The microscope is equipped with a Andor Zyla 4.2, a sCMOS camera with 4.0 MPixels with a big field of view and capable of maximum of 100 FPS at full frame (2048x2048).

    Location : UIC Room 4 (0B05), Bartolomeu Dias wing

    Microscope : Nikon Ti

    Year Installed: 2015

    Optics : Fluorescence and brightfield (with DIC)

    Camera : Andor Zyla 4.2 sCMOS 4.2Mpx

    Operating conditions : Temperature Controlled

    Keywords: screening, widefield, HCS, inverted microscope, high content screening


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 + GFP 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
Booking type
Regular reservation
Responsible
Unidade Imagiologia
Minimum booking time (in min)
30
Schedule tolerance (in days)
7
Detele tolerance (in hours)
0
Booking restriction status
Disabled

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
20x
PLAN APO λ
0.75
-
1
0.427
0.325
0.217
0.641
40x
Plan Fluor
1.3
Oil
0.2
0.246
0.162
0.108
0.370
100x
PLAN APO λ
1.45
Oil
0.13
0.221
0.065
0.043
0.331
20x
PLAN Fluor
0.75
MImm
0.35
0.427
0.325
0.217
0.641
10x
PLAN APO λ
0.45
-
4
0.712
0.650
0.433
1.068

(1) NA - Numerical Aperture. For more information, follow this link.

(2) These values were calculated using the Resolution formula adapted to confocals as seen in the Olympus micro website: R = 0.61*λ/NA

Installed filtersets

Filter
Excitation band
Dichroic
Emission band
Filter cube name
Filter position
Quad DAPI
BP 360/40
400/505/565/660
BP 470/40
Quadruple dichroic
1
Quad GFP
BP 480/40
BP 527/30
Quad TRITC
BP 546/12
BP 600/40
Quad Cy5
BP 620/60
BP 700/75
FITC
BP 480/40
505
BP 535/50
FITC
2
TRITC
BP 550/40
590
BP 610/50
TRITC
3
Cy5
BP 620/60
660
BP 700/75
Cy5
4

IMPORTANT: Take note that there are no individual filters in the microscope and all channels are done in a Quadruple Dichroic.

Spectra, lines & filters:

Transmissions stands for the % of light reflected, the higher the % the better.

Transmissions stands for the % of light reflected, the higher the % the better.

Transmissions stands for the % of light reflected, the higher the % the better.

Transmissions stands for the % of light reflected, the higher the % the better.

Transmissions stands for the % of light reflected, the higher the % the better.

Transmissions stands for the % of light reflected, the higher the % the better.

Transmissions stands for the % of light reflected, the higher the % the better.

Setup

Turn on procedures

  1. Turn on the first button (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 into the software using the USERS account.

    NIS Elements Log-in

  6. Set up your experiments on the microscope.

Turn off procedures

  1. If you are the last user of the day, close the acquisition software and turn off the computer. If you are NOT the last user of the day, you don't need to do the following steps.
  2. Turn off the microscope.
  3. Turn off the Camera.
  4. Turn off the Lamp LED button.
  5. If you used any immersion lenses be sure to clean them.

Acquisition

Acquiring images in a spinning disk confocal microscope is a complex procedure, first time users are required to contact the UIC staff for training. If you need assistance don’t forget to press “with assistance” at reservation time.

  • 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 and constant sample focus using the coverslip position as the reference, for a faster and less damaging focusing system compared to normal software ones. For more detailed information, follow the link or click the image to the right side.

  • Nikon PFS


Extra Info