1. Phase calibration pictures 3/9/10

2. Dark count image Required for: 1. determining image intensity in the absence of light. 2. also required for phase artifact correction

3. Background phase artifacts due to fixed optics Required for background subtraction of images collected with identical optics. Calibration procedure using a blank image -  raw image dark image [ [] - ] blank image These artifacts appear in all image data (upon close inspection) and can be removed

4. Application of correction procedure Removal of phase artifacts After correction

5. Properties of phase standard Correct ring alignment Mis-aligned rings Day to day instrument validation 10x, brightfield image PDMS stamp on microscope with ~20 µm high features

6. Benchmark with mis-alignment of phase rings Reduced contrast between features when alignment is incorrect

7. Cells with mis-alignment Cells with correct and incorrect alignment

8. Additional information from other imaging modes of the benchmark Correct alignment (PH1) and illumination Noisy illumination (room lights on) Contrast of benchmark affected Brightfield illuminationPH2 (not PH1) ring in illumination path