10:30 A – 11:30 PM
Project 2
- see notes
Probe condition optimization
- correctly identified sample types
- STORM laser powers may not be optimal, see observations under STORM
HPLC
- HPLC training with Bryan
STORM
- Finish O/N STORM2 run of BX-C BB3 sample
- STORM1 (PRISM2) training with Sang-hee. Recommended 50 mW 647 for STORM (measured at mounted power-meter location).
- built new chamber for repeated imaging of coverslip mounted samples (fits the center region of an 18mm round coverslip).
- STORM K-DNA in Kc cells.
- long / locally dense STORM works better when keep the density pretty high with the 405 and use no more 647 than necessary to get rapid switching (e.g. ~35% ATOF and 300 mW laser setpoint). Too much high laser and to little 405 and more dyes bleach before ever blinking, giving fewer localizations.
- Steve not properly configured yet to work on STORM1. Attempted overnight imaging aborted.
- Aborted K-DNA imaging on STORM2, move sample back to buffer, switch to doing Ph imaging on STORM2.
Ph
- imaging Ph in Ph-flag background (well, why don’t we image both)
- endogenous Ph seems to be inversely proportional to Ph-flag expression.
STORM Analysis
- Analysis of BB1 dots finished
- Analysis of BB3 dots started
- copying BB5 data to ProRAID
- Started analysis of BB5 data
- Preliminary analysis of B vs K DNA regions
Presentations
- did not manage to make much progress on presentations, too many experiments and data.