Fluorescence assays
Count cell fluorescence
Overview
Countess
™
3 FL Automated Cell Counter equipped with the optional EVOS
™
light cubes can be used for
a variety of fluorescent applications, including simultaneous counts of cells stained with two different
fluorescent dyes, GFP and RFP expression, and apoptosis and cell viability assays.
For instructions on installing EVOS
™
light cubes to your Countess
™
3 FL Automated Cell Counter, see
.
Count procedure
1.
Ensure that your fluorescent cell sample is homogeneously mixed.
2.
Load 10 µL of the fluorescent sample mixture per chamber into the sample slide as described
in “Load Countess
. Let the sample mixture settle for
30 seconds.
3.
(Optional)
: Press
Templates
to open the
Templates
screen and load the desired gating parameter
as described in “Load a template”
.
4.
Insert the sample slide into the slide port (“Exterior instrument parts”
), making sure that
the sample side is inserted completely into the instrument. You will hear a soft click if the slide is
pushed in correctly.
5.
When the slide is inserted, the instrument automatically illuminates the sample, sets the intensity of
brightfield illumination, and auto focuses on the cells.
Note:
Auto lighting is optimal for producing accurate results, but if needed, can be disabled by
adjusting the lighting slider.
6.
To view your sample under a different light source, press the desired light source button (
BF
(brightfield) or a source provided from an installed light cube (e.g.,
GFP
,
DAPI
,
TX Red
, etc.)). The
instrument displays the sample in the selected channel (brightfield or fluorescent).
In the example below, the sample is displayed in the DAPI channel.
6
30
Countess
™
3 FL Automated Cell Counter User Guide
Summary of Contents for AMQAF2000
Page 73: ......
