4 um Tetraspeck beads used for all images on this page. Stuck to Zeiss high performance coverslip with poly-L-lysine and mounted in Prolong Gold.
All imaging with excitation laser (or LED) in range 633 to 647 nm with detection in 640 to 760 nm range.

LSM 700, Axio Imager 2
Plan-Apochromat 40x/1.4 Oil DIC M27
PMT
Pixel size 0.1041950 um and Z step 0.2 um.

Z series were collected with pinhole set to 1 Airy unit for confocal and max open for widefield.

From left to right, widefield XZ sections with raw data and with contrast adjusted per XY slice before XZ section and at right confocal versions.

 

LSM 880, AxioObserver
Plan-Apochromat 63x/1.4 Oil DIC M27
GaAsP detector
Pixel size 0.1054306 um and Z step 0.22 um.

We imaged two different beads in different fields and both had this odd double appearance. We need to repeat this.

 

Nikon CSU-W1 spinning disk
Apo 60x/1.4 Oil λS DIC N2 MRD71600
Andor DU-888 X-12209
Original pixel size 0.2166667 um and Z step 0.22 um; scaled 2X in XY for display here.

 

Nikon Ti (widefield only)
Plan Apo60x/1.4 Oil Ph3
ANDOR Neo/Zyla 5.5
Pixel size 0.1083333 um and Z step 0.22 um.

(The horizontal lines are a problem with light instability or on/off timing discovered in this dataset that we need to address.)


Nikon Ti (widefield only)
100x/1.49 Oil DIC TIRF
ANDOR Neo/Zyla 5.5
Pixel size 0.065 um and Z step 0.22 um.

 

Zeiss Elyra 7
Plan-Apochromat 63x/1.4 Oil DIC M27 at RT
Images rescaled for display here.

(The vertical line is dust on the camera discovered in this imaging session that needs to be addressed.)
More pictures of this bead including 3D reconstruction here.

 

 

last updated 20200804