RCF Microscopy Manual
5/2/2017
Authors Cory Nook and Chaowei Shang
Page 7 of 19
To illuminate your sample with fluorescent light,
press the header icon labeled Reflector to pull up the
fluorescent filters. On the right sidebar, make sure
that the Transmitted light (TL) Illumination is off and
that the Reflected Light (RL) Illumination is on.
Filter choices are labeled as, on the TFT display:
34 BFP
(EX 390/20, EM 460/60) – for DAPI, Hoerchst, AMCA
38 HE GFP
(EX 470/40, EM 525/50) – for GFP, FITC, and
prevents red emission bleed through
31 AF 568
(EX 565/30, EM 620/60) – for Cy 3, Texas Red
17 AF 488
(EX 485/20, EM 540/25) – for GFP, FITC, and
prevents red emission bleed through
50 Cy 5
(EX 640/30, EM 690/50) – for Cy 5 and far red dyes
3.
MOUNT AND VIEW THE SAMPLE THROUGH THE MICROSCOPE
If you are using an immersion objective, place a VERY small drop of the appropriate immersion fluid on the
objective face, or the coverslip. Be sure to only use the immersion fluids located next to the microscope. If you
accidentally use the wrong fluid, please contact staff right away. If you are using a slide, insert it coverslip
down into the specimen stage template. Below is a table of our current objectives, imaging medium, and
brightfield capabilities.
