1. Project Scope Shootout Stella Breslin Biosc 3 / Cohort 3 Spring 2010 Slide = Chroma Full Skin Section Nuclei stained with Hoechts F-Actin - Alexa 488 Elastin - Cy3 Collegen - Cy5

2. Chroma Leica Sp5 5/4/10 5/4/10 SBChromaMay4.lif/Image051 mag: 20x Plan Apo 2channels 512x512 AOTF 30% (514) PMT2 519nm - 630nm off 0 HV 827 / AOTF (514) 20% PMT3 638nm - 765nm off -1 HV 750 / line avg 1 frame avg 3 pinhole 30.3 µm pinhole airy 0.5

3. Leica Spinning Disk 5/15/10 5/15/10 ChromaSD.lif/Series22 HC PL FLUOTAR 10.0x0.30 DRY 3 Channels 1 frame Exp G 932 ms, Red 932 ms, Blue 1000 ms Gain: G 5.9 R 5.9 B 6.3

4. Nikon 20x 5/8/10 Nikon 3 5/8/10 Mag: 20x Exp: TRITC 200ms FITC 500ms DAPI 52ms Contrast Stretch ImageJ

5. Zeiss 2 / DAPI Filter Added 5/11/10 ChromaSlide Multichannel mag: 20x Exp: FITC 346ms Rhod 286ms DAPI 145ms Br -.057,-0.57-,0.66 Con 1.19, 1.06, 1.37 gamma 1.0

6. Zeiss 2 / Darkfield 5/11/10 ChromaSlide Darkfield mag: 20x Exp: 130ms Br -0.03 Con 1.6 gamma 1.0

7. Chroma -Zeiss 2 / Rhod/FITC/DAPI/DIC overlay - 5/11/10 ChromaSlide MC Overlay Mag: 40x Exp: FITC 598ms Rhod 286ms DAPI 24ms Con 1.0 gamma 1.0 DIC exp 4ms / Contrast stretch each channel and DIC overlay done in Adobe PS

8. Chroma -Olympus 3 - 5/22/10 ChromaSlide Multichannel mag: 10x MC Exp: Red 518.5ms Green 441.9 Offset R -894 G-662 Gain 1.0 Color Merge Contrast Stretch in Adobe PS

9. Chroma - Olympus 3- 5/22/10 5/22/10 ChromaSlide mag : 10x Phase exp: 18.6 ms gain 1.0 offset 0 Contrast Stretch Image J

10. Comparisons Slide 6: your succinct comments comparing the images acquired on