4 um TetraSpeck beads stuck to #1.5 Zeiss high quality coverslip with poly-L-lysine and mounted in Prolong Gold set for 3 days. Imaged with 642 nm laser. Zeiss Elyra 7.
Plan-Apochromat 63x/1.4 Oil DIC M27 at RT with matching oil
Other image metadata here.
This was collected with Lattice SIM so each Z slice shown as raw data bead is actually median projection image of 13 phases. (Remapped hyperstack from xyczt to xyctz and then median projected.)
Slices through the Z stack rescaled smaller for webpage presentation.
Next picture is XZ reslices through bead near center, but not exact diameter.
Left: raw data.
Center left: default SIM reconstruction settings.
Center right and Right: Before resampling the stack in Z axis, the float command was applied to adjust contrast per slice.
One bead was cropped out in XY and sliced in Z axis and scaled (4.8x in Y) for orthagonal pixels. 3D projection with ImageJ with transparencies set to 50 and 25.
Before resampling the stack in Z axis, the float command was applied to adjust contrast per slice.
I expected the bead surface closest to the lens to be round and flat and the bead surface farthest from the bead to be conical.