Details for sorting your sample
1) Contact:
name
Group
E-mail or phone
2) When is your preferred day?
3) The FACSAria II SORP has this Configuration:
Laser
UV Laser
(355nm)
Filter
450/50
530/30
Color
DAPI
Indo1
(blue)
exam
ple
Hoechst
33258
Emerald
300
Violet Laser
(405nm)
Blue Laser
(488nm)
450/50
525/50
525/50
Alexa
405
AmCyan
FITC
Pacific
Blue
Pacific
Orange
Alexa 488
GFP
YellowGreen Laser (561nm)
695/40
585/15
610/20
660/20
PerCp
PE
PE-Texas
Red
PE-Cy5
mCherry
mPlum
PerCpCy5.5
DsRed
710/50
YGPerCpCy5-5
Red Laser (640nm)
780/60
670/14
APC
PE-Cy7
PE-Cy5.5
Alexa
647
a) Put the used dyes in the right column of the table.
b) You can only put in one color per column!
E.g.: Rho 123: it is excited with the blue laser and the emission will be detected in the filter 525/50nm, so put
Rho 123 in the column like FITC
c) If the excitation and/or emission are unknown put the color name here:
write the Excitation max.
and Emission max.
730/45
form the protocol
780/60
APC-H7
Alexa
700
APCCy7
4) Sample, average cell size.
cell size:
5) To optimize the sort time, answer the question, please.
a) How many cells should be sorted?
b) What is the percentage of your populations?
(Note that you can’t sort more than 4 different populations at the same time!)
d) What is your goal?
6) Does your sample need a certain temperature?
%
%
%
%
Purification ?
no
7) You should bring along:
a)
b)
c)
d)
8) Remark:
Your stained/transfected and filtered sample.
Tubes or pates for the cell to be sorted into, filled to >1/3 with growth medium.
An unstained sample or for cells transfect with GFP or other fluorescent protein an untransfect and unstained sample.
If you use more than one color, bring compensation controls with you (for that you can use your cells or beads).