NEW ACCOUNT SETUP

1. Double click on top bar of Elements
2. General tab -> uncheck “save changes to optical  configuration automatically”
3. Appearance tab -> uncheck “initial zoom 100% for Live and Capture”
4. Layout tab -> open latest “layout” file in data-drive base
   1. Sort by date modified most recent and click on the file xxx_layouts.xml
   2. Import and Replace
   3. Click “do this for all future”
5. Device manager -> settings -> uncheck box by warning for switching paths
6. Calibration -> optical configurations
   1. Import most recent OC file on drive drive (xxx_OCs.xml)
   2. Window “import objectives” click Yes
7. Devices -> Set Z-device to Ti2 drive (NOT PIEZO)
   1. Devices -> Enable mouse joystick
8. ND Acquisition -> z-stack -> change z-device to Ti2 drive

TOPIC TO HIT

• Light path (Camera vs Eyepiece)
• Histogram vs LUTs table and saturation/pixel saturation indicator
• Optical Configurations
• ND Aquisition 
  • Lambda 
  • Large Image
  • Multipoint
    • Oil objectives
  • Z-stack
    • Movies
  • Time Lapse 
• How to save images
  • ND2 vs other types 
    • How to recall ND2 meta data

1. Double click on top bar of Elements
2. General tab -> uncheck “save changes to optical  configuration automatically”
3. Appearance tab -> uncheck “initial zoom 100% for Live and Capture”
4. Layout tab -> open latest “layout” file in data-drive base
   1. Sort by date modified most recent and click on the file xxx_layouts.xml
   2. Import and Replace
   3. Click “do this for all future”
5. Device manager -> settings -> uncheck box by warning for switching paths
6. Calibration -> optical configurations
   1. Import most recent OC file on drive (xxx_OCs.xml)
   2. Window “import objectives” click Yes
7. Devices -> Enable mouse joystick

TOPIC TO HIT

• Light path (Camera vs Eyepiece)
  • Nikon A1
  • 1st Eyepiece all the way on the left
• Histogram vs LUTs table and saturation/pixel saturation indicator
• Optical Configurations
• ND Acquisition
  • Lambda
  • Large Image
  • Multipoint
    • Oil objectives
  • Z-stack
    • Movies
      • mp4
  • Time Lapse
• How to save images
  • ND2 vs other types
    • How to recall ND2 meta data

NEW ACCOUNT SETUP

1. Double click on top bar of Elements
2. General tab -> uncheck “save changes to optical configuration automatically”
3. Appearance tab -> uncheck “initial zoom 100% for Live and Capture”
4. Layout tab -> open latest “layout” file in data-drive base (drive letter varies)
   1. Sort by date modified most recent and click on the file xxx_layouts.xml
   2. Import and Replace
   3. Click “do this for all future”
5. Calibration -> optical configurations
   1. Import most recent OC file on drive (xxx_OCs.xml)
   2. Window “import objectives” click Yes
6. Devices -> Set Z-device to Ti drive
   1. Devices -> Enable mouse joystick
7. ND Acquisition -> z-stack -> change z-device to Ti drive

NEW ACCOUNT SETUP

1. Follow import settings instructions on the desktop.
2. Go over filter cube removal and replacement.
3. Go over removing only the DAPI cube for BF/Phase and replaced when done.
4. Drive home that cubes should be checked before using.
5. Drive home that Channel 1 is DAPI, 2 GFP, 3 RFP, 4 Cy5.

When Booking 

• Mention that they should use glass bottom 35-mm petri dishes as a normal wet mount might not allow you to move around when in an oil objective.
• We sell for $28 for a 10 sleeve or they can buy online

DIC (if they need DIC) 

• Better for if you need anything quantitate about your data
  • Normally for people doing anything other than bacteria
• Explain Cohler and set up
  • Explain in 10x
  • Focus on sample first
  • Close aperture
  • Sharpen the light with the two knobs
  • Open back up aperture
  • Turn on polarizer by pushing it over
    • Adjust polarizer by turning the knob next to Lense
      • Normally far right is best
  •  Mess with bottom aperture for out of focus light
• Switch to DIC OC
  • Make sure that you are in an objective that has the DIC prism or else it won’t work
    •  60x or the farthest to the right 100x
  • Mess with n if needed
    • It’s good to switch through each one quickly to make sure you are on the best one for their sample
      • N1- For higher mag, Should be what you need
      • N2- For lower mag, normally bacteria is not really visible at lower mag but can try
      • Nr-
• Explain DIC
  • When the light comes down from the top any photon that does not touch another substance, like your sample, will appear dark/black. Any photon that does touch your sample will show as white or a shade of gray depending on the angle that the light is reflected with.

PHASE (if they need PHASE)

• Better  if you just want to count your cells
  • Normally for people doing bacteria (Microbiologists)
• Explain Kohler and set up
  • Explain in 10x
  • Focus on sample first
  • Close aperture
  • Sharpen the light with the two knobs
  • Open back up aperture
  • Turn on polarizer by pushing it over
    • Adjust polarizer by turning the knob next to Lens
      • Normally far right is best
  •  Mess with bottom aperture for out of focus light
• Switch to phase OC
  • 100x phase
  • Mess with P if needed
    • It’s good to switch through each one quickly to make sure you are on the best one for their sample
      • Phase 3 is normally the best
• Explain DIC
  • There are two opposite filters, one above and one below that are inverted of each other
  • The light is inverted because anything that the light doesn’t interact with gets blocked

TOPIC TO HIT

• Light path (Camera vs Eyepiece)
  • Prime for camera
  • Manual switch on side of microscope for switching between the two
• Histogram vs LUTs table and saturation/pixel saturation indicator
• Optical Configurations
• ND Acquisition
  • Lambda
  • Large Image
  • Multipoint
    • Oil objectives
  • Z-stack
    • Movies
      • Save as MP4
  • Time Lapse
• How to save images
  • ND2 vs other types
    • How to recall ND2 meta data

ACCOUNT SETTINGS

1. Double click on top bar of Elements
2. General tab -> uncheck “save changes to optical  configuration automatically”
3. Appearance tab -> uncheck “initial zoom 100% for Live and Capture”
4. Layout tab -> open latest “layout” file in data-drive base
   1. Sort by date modified most recent and click on the file xxx_layouts.xml
   2. Import and Replace
   3. Click “do this for all future”
5. Calibration -> optical configurations
   1. Import most recent OC file on drive (xxx_OCs.xml)
   2. Window “import objectives” click Yes
6. Devices -> Set Z-device to Ti2 drive
   1. Devices -> Enable mouse joystick
7. ND Acquisition -> z-stack -> change z-device to Ti2 drive
8. IMPORTANT:  Explain ways to get help – new chat to LMFStaff on Teams, website pop up with chatbot, find human in S531/529.