Question: Interpret The Result: 0.269 0.072 CD4. T Cells 7.5 CD4’CD8′ T Cells 0.813 4183.1 WT WT 5.9) 34,61 0.067 0.078 2.82 84.9 0.101 Crtam CD8 Crtam 2.8 32.31 -IFN-Y- 802 0.121 3.91 Cadm1 15.85 CD4 CD4- 3-33 Calculate The Stokes Shift For Each Fluorophore In The Table: Stokes Shift Excitation (nm) Fluorophore Alexa488 Emission (nm) 525 488 650 660 APC 675 …

Question: Interpret The Result: 0.269 0.072 CD4. T Cells 7.5 CD4’CD8′ T Cells 0.813 4183.1 WT WT 5.9) 34,61 0.067 0.078 2.82 84.9 0.101 Crtam CD8 Crtam 2.8 32.31 -IFN-Y- 802 0.121 3.91 Cadm1 15.85 CD4 CD4- 3-33 Calculate The Stokes Shift For Each Fluorophore In The Table: Stokes Shift Excitation (nm) Fluorophore Alexa488 Emission (nm) 525 488 650 660 APC 675 …

Interpret the result: 0.269 0.072 CD4. T cells 7.5 CD4CD8 T cells 0.813 4183.1 WT WT 5.9) 34,61 0.067 0.078 2.82 84.9 0.101

Calculate the Stokes shift for each fluorophore in the table: Stokes shift Excitation (nm) Fluorophore Alexa488 Emission (nm)

Fluorophore Excitation (nm) Stokes shift (nm) А Diameter (nm) Quantum Yield 488 97 0.93 10 B 488 37 5 С 0.82 405 146 10 0.35

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Interpret the result: 0.269 0.072 CD4. T cells 7.5 CD4’CD8′ T cells 0.813 4183.1 WT WT 5.9) 34,61 0.067 0.078 2.82 84.9 0.101 Crtam CD8 Crtam 2.8 32.31 -IFN-Y- 802 0.121 3.91 Cadm1 15.85 CD4 CD4- 3-33 Calculate the Stokes shift for each fluorophore in the table: Stokes shift Excitation (nm) Fluorophore Alexa488 Emission (nm) 525 488 650 660 APC 675 694 Cy5.5 FITC 488 525 488 585 PE 403 455 Pacific Blue 595 613 Texas Red Which fluorophores could NOT be used on a flow cytometer with only a 488 nm excitation light source? Which pair of fluorophores could NOT be used together in the same experiment? Fluorophore Excitation (nm) Stokes shift (nm) А Diameter (nm) Quantum Yield 488 97 0.93 10 B 488 37 5 С 0.82 405 146 10 0.35 D 488 15 279 0.74 E 640 10 35 0.82 F 488 25 37 0.97 You are setting up a flow cytometry experiment to stain cells for the following 3 antigens: Antigen 1 – Highly expressed transcription factor Antigen 2-Weakly expressed cell surface protein Antigen 3-Strongly expressed cell surface protein Your flow cytometer has excitation light sources of 488 nm and 640 nm, and it can detect emissions of 525 nm, 585 nm, and 675 nm. Which fluorophore should you use with each antigen?