8-color MACSQuant Acquisition Request Form Version 1 (effective 1 Aug 2012)
It is strongly recommended that you consult the IML Director (Jacqueline.smith@dartmouth.edu) to
optimize your staining cocktails before carrying out staining experiments.
Charges:
8-color MACSQuant (7 Rubin) (operator-assisted)
7-color MACSQuant (operator-unassisted)
8-color MACSQuant (5 Remsen) (operator-unassisted)
$75/hour
$50/hour
$75/hour
($150/hour Company rate)
($100/hour Company rate)
($150/hour Company rate)
If PI is used as a stain, machine requires a full bleach clean before the next client, requiring a 20
minute additional charge.
Operator assistance includes setting up your samples, labeling fields, and preparing prints of data if
requested. Tech time is billed in 15-minute blocks but may be waived for initial consultation to
develop settings. 8 color MACSQuant on 7 Rubin is operator assisted only.
Lab PI ____________________ Phone #______________
Requestor___________________
Today’s Date ______________ Scheduled date of acquisition _______________
Account string to be charged_____________________________________________
Species:
___ human
___ mouse
__________________ (describe)
Cells:
___ PBMC
___ lysed whole blood
___ spleen
__ MLN
Other (describe) _________________________
Sample Prep: Check all that apply. BSL2 samples MUST be fixed for aerosol safety.
___ unfixed
___ fixed
___ extracellular stained
___intracellular stained
Cell concentration: ________________/ml
Total number of samples: _______
Volume of each sample (l): ___________
Compensation and setup controls
Compensation requires single fluorophore-stained beads and/or cells.
What was stained?
_____ cells _____beads
Please supply an unstained sample of compensation beads and/or cells.
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8-color MACSQuant Acquisition Request Form Version 1 (effective 1 Aug 2012)
What antibodies and colors were used for each PMT? State ‘none’ for unused PMTs
V1
V2
B1
B2
B3
B4
R1
R2
Unstained sample
Flow rate:
____ slow (very rare events at moderate cell concentrations). Allow ~10 minutes/150 l sample;
_____ medium (suitable with cells up to 1.5 e^7/ml). Allow ~6 minutes/150 l sample;
_____ fast (high frequency events with cells <1e^6/ml where high resolution unnecessary)
Allow ~2.5 minutes/150 l sample.
PI requires 2 bleach clean cycles after use, so please schedule an additional 20 minutes. When
sample cell concentration is >1 e^6/ ml then automated mixing is required and takes an extra 30 sec
per sample.
Samples should be filtered by customers (70  mesh) before acquisition. If the machine becomes
clogged, users will be charged 20 minutes extra time for a backflush cycle. Impacted clogs that
cannot be cleared with the backflush cycle will result in additional charges.
Estimated acquisition time: _______ minutes (to be completed by customer)
___________________________________________________________________________
Following section to be completed by technologist preparing acquisition
Time begun
__________________
Settings Name
__________________
Layout name
__________________
Time ended
File name first sample __________________
last sample
2
__________________
_________________
8-color MACSQuant Acquisition Request Form Version 1 (effective 1 Aug 2012)
Prior to acquiring samples, acquire an aliquot of 8 peak beads on your channel voltages with
FSC/SSC set to 475/430 Lin. Users can compare 8 peak beads file from experiment to experiment to
evaluate instrument performance. This also provides a benchmark of resolution at the user preferred
voltages.
File name 8 pk beads_____________
Parameter
voltage
scale
FSC
SSC
B1
B2
B3
B4
V1
R2
R1
R2
Trigger
Compensation tubes: Was there an adequate stain index for each fluorochrome?
____________________________________________________________________
Sample quality:
Does viability appear adequate?
___________
Does the staining appear adequate?
___________
Was there hemolysis for whole blood samples?
___________
Are the samples ‘clumpy’ ?
___________
If quality of sample is in question, does the requestor understand the limitations of the data
provided____________.
Had the antibodies been titrated? __________________
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8-color MACSQuant Acquisition Request Form Version 1 (effective 1 Aug 2012)
IF plates are used and samples are identified other than by location in plate provide
identification in the following format:
1
2
3
4
5
6
7
8
9
10
11
A
B
C
D
E
F
G
H
4
12