SICKKIDS-UHN FLOW CYTOMETRY FACILITY

ASSISTED FLOW CYTOMETRY & DATA ANALYSIS FORM

Name / Contact information
PI: / Location:
User: / Email:
Experiment Date: / Phone #:

Instrument Selection & booking: is made by FCF staff based on your experiment and biosafety requirements

Acquisition time required: depends on # of samples and events recorded. Min of 30 minutes is required for instrument setup and cleaning.

CELLS & CELL PREP INFORMATION
1. Species origin of cells (e.g. Human, mouse)
2. Tissue Source (e.g. spleen, lung)
3. Primary or cell line (if cell line provide name)
4. # of cells per sample
5. Estimated frequency of population of interest?
6. Buffer used to re-suspend cells?
7. Are these cells adherent?
8. Cell size (~um)
9. Are these cells prone to clumping?
10. # of events to acquire?
11. Was a viability stain used (if yes, please specify)
12. Are these cells expressing any fluorescent proteins (e.g. RFP, GFP)
13. Risk Group of samples (RG1,RG2,RG3 or RG4)
14. List any chemical treatments (e.g. PMA, Ionomycin) and associated risk (e.g. carcinogen)
15. List any infectious biological agents cells have been exposed to (e.g. bacteria, retrovirus, EBV)
16. Do cells/biological agents require Level 3 operating procedure as determined by Sickkids/UHN Research Biosafety Committee ? If yes, identify cells/biological agent(s) and Risk Group
17. Have the cells been infected or transfected with any gene transfer vector, and if yes, which one?
18. Are cells fixed? (Yes or No)
If yes, list fixative if it’s not the recommended (0.5-2% PFA for at least 30’)
SAMPLES MUST BE FILTERED IMMEDIATELY PRIOR TO RUNNING (100uM size filter)
FCF can only use 5ml FACS tubes for acquisition.
(Corning Inc, cat # 352052, 5ml Polystyrene Round-Bottom tubes)

CONTROLS & EXPERIMENTAL SAMPLES

NAME / Marker 1 + Fluorochrome / Marker 2 + Fluorochrome / Marker 3 + Fluorochrome / Marker 4 + Fluorochrome / Marker 5 + Fluorochrome / Marker 6 + Fluorochrome / Viability
Example / wtSpleen Untreated / pSTAT5 PE / pLCK Ax647 / CD4 eF450 / CD8a PE-Cy / TCRab PerCP-Cy5.5 / B220 NC605 / DAPI
CONTROL 1 / UNSTAINED / x / x / x / x / x / x / x
CONTROL 2
CONTROL 3
CONTROL 4
CONTROL 5
CONTROL X
SAMPLE 1
SAMPLE 2
SAMPLE 3
SAMPLE 4
SAMPLE 5
SAMPLE 6
SAMPLE 7
SAMPLE 8
SAMPLE 9
SAMPLE 10
SAMPLE 11
SAMPLE 12
SAMPLE 13
SAMPLE X