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This page contains various protocols for different instruments that you may find in the instruments page.

Nikon TIRF

Laser adjustment

Before you start:

Manual parts to check

  1. Position of microscope magnification lens
  2. If using Andor DU897 camera the position of the 3D STORM lens
  3. Correction collar of objective (temperature, cover glass thickness)
  4. Insert filter cube into filter wheel (if not currently available)
  5. Check position of ND filters in front of 405, argon and 561 laser heads (probably the up position)
  6. Laser arm:
    1. Position of magnifying lens (up=TIRF, down=STORM)
    2. lamda plate and nd filter positions
    3. Angle and collimation of laser (spot on roof)

 

Check laser collimation

  1. Focus on sample using transmitted or epi light
  2. Open environmental chamber roof and push transmitted light arm to back position (check CO2 is off)
  3. Adjust laser angle offset from software to directly upward, depending on cube should be approx 3200
  4. Adjust point on roof
    1. loosen collimator lock screw
    2. micrometer screw for y-direction
    3. software offset for x-direction
    4. collimator push/pull for spot size (small round spot with concentric circles around)
    5. tighten collimator lock screw

Nikon N-STORM protocols:

NEW N-STORM_PROTOCOL_4_4_2013.pdf

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