Leica SP8 Confocal Quick Start Manual

1.

Initial Set-up

~ Log in to NUCore and start a reservation.
~ Turn on power strip to turn on all components (Computer, Scanner power,
Laser power, Laser key). Note: Do not turn off the laser power key by itself!!
~ Log into computer, if necessary (password: user).
~ Launch LAS software. Select Resonant Scanning, if applicable.
~ Ensure 10x objective is in place and say "OK" when dialog appears.
~ Navigate to Laser panel (A) and select "classic" or "express" view for
your convenience.
~ Put slide on scope and set up image acquisition.
2.

Mount sample on microscope

~ Place slide on stage 'coverslip down'.
~ For transmitted light imaging: Select BF button and press TL-Shutter on LED screen. Focus on a sample.
~ For fluorescent light imaging: Select FLUO, then filter cube from buttons on LED screen, open TL-Shutter.
~ Focus on sample and find cell/region of interest. Close shutter.
3.

Confocal imaging

~ In software, navigate to
"Acquire" (B)
tab .
~ To set Beam Path Settings,
first activate the laser(s)
needed by clicking "ON" and
"+" (C). Set laser intensities at
0.5-2% to start.
~ Activate the HyD detector(s)
or PMT(s) by selecting the
"ON" button(s) (D). Choose a
color for the detector display
by clicking the color bar (E)
and selecting a LUT from the
window that opens.
~ To display emission peak to
guide detector settings,
choose from list of common
fluorophores (F).
Leica SP8 Confocal
Quick Start Guide