Spinning Disk W1 NIKON troubleshooting

  • Check on the NIS-Element software that “Eyepiece-EPI” is selected for the fluorescent marker, otherwise select “Eyepiece-DIA” for brightfield illumination.
  • For the fluorescent marker, select a filter (“DAPI”: Blue; “GFP”: Green; “CY3”: Red) and check that the fluorescent lamp shutter is open.
  • Go up the lenses by turning the wheel towards you.
  • When the microscope is in focus, it beeps.
  • Check on the software that « Spinning » is selected, to direct fluorescence towards the spinning head and camera
  • Click on « LIVE »
  • Click on « AUTOSCALE » in the live windows or adapt the dynamique in LUT’s windows
  • Increase or decrease the exposure time and the power of laser
    • 385 nm for blue fluorochrome
    • 475 nm for green
    • 550 nm for red
    • 621 nm for far red
  • Clean lenses with kimtech and ethanol.
  • Check that the slide has been turned over and that the coverslip is in contact with the objective.
  • And refocus
  • In the software search bar, type :
    • “Acquisition” : To display the acquisition window (Live, Eyepiece, Lenses, Time exposure… )
    • “ND acquisition” : To set your acquisition parameters (Multi-Wavelength ; Z-stack ; Timelapse ; Multiposition… )
    • “LUT’s” : To display the histogram of image dynamics
  • Check in the first drawer next to a laboratory bench or next to the microscope
  • Remove the holder and install it
  • Check the ROI status : if it is ticked, a region of interest will be applied on the image. Ensure to untick it.