Finalized light-sheet design and more drosophila imaging

October 10, 2019

by Abhijit Marar

Drosophila imaging:

Our collaborator Daichi Kamiyama purchased new CF568 dye which has a higher probability of attaching the primary antibody. This has allowed for better imaging of the myopodia. In addition to this, we have been trying to perform two-color sequential imaging of the growth-cone (hrp- AF647) and Fas2 protein present in the growth cone and have had some promising results. Shown below are two images of the Myopodia stained with the new CF 568 dye.

STORM imaging of the Myopodia labeled with different concentration of CF568 (left) Concentration of secondary antibody is 1:500, precision is 12 +/- 7 nm (right) concentration of secondary antibody is 1:200, precision is 22 +/- 6 nm.

 

The images below show two-color sequential imaging of the growth cone labeled with HRP-AF647 and Fas2-CF568

(left) STORM image of HRP-AF647 (center) STORM image of Fas2_CF568 (right) Composite image with two images overlayed on each other after drift correction. Average Precision is 21 +/- 6 nm

 

(left) STORM image of HRP-AF647 (center) STORM image of Fas2_CF568 (right) Composite image with two images overlayed on each other after drift correction. Average Precision is 23 +/- 7 nm

 

Finalized light-sheet design:

Below is the final design of the light-sheet add-on to the STORM setup. We are currently in the process of buying the parts to assemble it.