Grishchuk Lab

Protocols and Software

EXPERIMENTAL PROTOCOLS:

1. Manufacturing reusable flow chambers.
2. Preparation of coverslips.
3. Preparation of GMPCPP-stabilized microtubule seeds.
4. Attachment of microtubule seeds to the coverslips.
5. Preparation of segmented microtubules. 
   Video illustrating protocols 1-5 (Volkov et al., 2014)
   
6. Experimental observation of the protein tracking with depolymerizing microtubule ends.
7. Preparation of streptavidin-coated beads
8. Preparation of unevenly coated beads
9. Recording the photobleaching kinetics for the fluorescently labeled protein molecules.

IMAGE ANALYSIS

1. Correcting for unevenness of illumination.
2. Determining the intensity of single fluorophore.   
3. Quantification of bead brightness Protocol | Source Code.

SOFTWARE

1. CENP-E tip tracking (Gudimchuk et al., 2012) Manual | Source Code
2. Molecular-mechanical model of the dynamic microtubule (Zakharov et al., 2015) Manual | Source Code
3. Tethered Cargo Motion (TCM) Monte Carlo simulation software (Gudimchuk et al., 2018) Manual | Source Code