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1 TM Gallios Gallios 4 Laser System Powerful Versatile Performance Fast Track to Success CE E

2 Gallios High End Flow Cytometer 1 2

3 Accuracy Log Decade Display Gallios Power(mV) Blue loss Red loss Violet loss Blue power Red power Violet power Gallios FCM A FCM B Yellow loss Yellow power 3 4

4 Flexibility 0.5m NOISE 0.4m FS 638nm 561nm 488nm 405nm APC APC AF700 APC AF750 FITC PE ECD PC5 or PC5.5 PC7 Pacific Blue Krome Orange 488/561nm This Dimension should = Focal Distance of Lens (1 unit) to maintain Fourier Plane response. GFP mcherry PerCP PC7 Photo-Sensitive Devices Wide 8-19 Narrow 1-19 Beam Dump Blocking Laser Beam Divergence 2 Focal Length of Lens = 1 unit Max = 19 Max = 19 decreasing Intensity of Scattered Light as the Amgle increases Wide 8-19 Narrow 1-19 Core Stream / Laser Intersection Point Wide 8-19 Lasers Channels FL1 FL2 488 nm 561 nm FL3 Fluorochromes Fluorescein PE ECD Fluorescent Proteins & Viability GFP dtomato mkate mcherry dsred mkate FL4 PC5 PC5.5 FL5 PC7 FL6 APC 638 nm 405 nm FL7 APC A700 FL8 APC A750 Fixable efluor 780 FL9 Paci c Blue FL10 Krome Orange 5 6

5 Automate FL2 INT LOG FL2 INT LOG FL1 INT LOG FL1 INT LOG FL2 INT LOG FL1 INT LOG 7 8

6 High Performance Singlets Singlets Singlets Singlets DyeCycle Violet Blue DyeCycle Violet Blue Panel A E. coli Panel B E. coli FL1 (525/30) Panel C E. coli DRAQ7 DRAQ7- FS INT LIN Cell CD25-PE Treg 0.7% FS-W FS-A AUTOFL FS-A Channel FL1 FL2 FL3 FL4 FL5 FL6 FL7 FL8 FL9 FL10 K : 4.91% FL4-LinPC5 Fluorochrome FITC PE ECD PC5 BS-PC7 APC APCA700 APCC7 DAPI Lin DAPI Log B : 96.63% C : 0.67% FL2-CD133PE Specificity CD38 CD133 CD34 CD14,33,20,3, GlyA VEGFR2 CD31 CD146 CD45 DNA DNA H : 0.71% I : 0.61% J : 0.45% FL10-DAPI Log FL1-CD38 FITC FL2-CD133PE FL1-CD38 FITC GTE 2N LinP CD45P LinP CD34P Clean CD34P E: 90.06% G : 0.72% F: 95.56% FL8-CD45AC7 FL3-CD34ECD FS-A FL1-CD38 FITC CD45N LinN LinN CD34P L L L L : 85.56% FL6-CD31APC M1:0.00% M3:0.00% D: 0.07% M2:100% FL3-CD34ECD FS-A FL1-CD38 FITC FL2-CD133PE FL5-VEGFR2PC7 Identifies Committed Heme Progenitor w/cd34 Heme Progenitor Subset Heme Progenitor Heme Lineage Endothelial Progenitor Mature Endothelial Endothelial Progenitor Leukocyte Cell cycle Debris M4:0.00% FL3-CD34ECD FL7-CD146AA700 FL6-CD31APC FL2-CD133PE FL3-CD34ECD M2:2.55% N1:16% 01:98% M3:74.84% M4:6.69% M2:2.55% 03:0.00% M4:0.00% 0.73% 0.00% DyeCycle Violet Red DyeCycle Violet Red FL2 (582/15) FL2 (582/15) Panel D Panel E E. coli Mouse spleen FL2 (582/15) FL2 (582/15) kawa9.27 cellcycle PI-stnd-01 A FL3 Lin.HST 7-AAD Lin FS CD62L-BV421 CD62L-BV421 A Gate 85.3% C Gate 71.7% CD8-BV510 Single Cell A 3.1% SS INT LIN B 72.9% D 9.7% C 8.9% CD4-FITC Cyclin A2 FITC CD3-PC7 B Gate 95.7% CD62L-BV421 CD62L-BV421 G2 Phase 22.5% M Phase 26.7% D Gate 71.9% G0-G1 76.2% 7-AAD PEAK 7-AAD FS PEAK SS PEAK S 13.9% G2-M 10.0% Cell Number DNA Content 9 10

7 Support CD CHART

8 13 14

02

02 54 163116831 02 1 168 54 158 53 162 53 148 52 152 52 10,000 0 40,000 30,000 20,000 50,000 70,000 60,000 1,000 500 1,500 2,000 0 2,500 3,000 4,000 3,500 4,500 168 54 158 53 162 53 148 52 152 52 03 52148

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