Monitoring Transcription Factor Oligomerization in Single Living Cells by Number and Brightness Analysis
One key step in the activation of inducible transcription factors is their homooligomerization, which can be measured in individual living cells by a fluorescence microscopy technique called Number and Brightness analysis (N&B). In this chapter we describe how to acquire and analyze confocal microscopy time-series to provide information about transcription factor oligomerization in living cells using this technique.
Key wordsNumber and brightness Oligomerization Transcription factor Live cell imaging Image analysis
We are grateful to Dr. Valeria Caiolfa and Dr. Moreno Zamai for their support on N&B analysis. Confocal imaging was carried out in ALEMBIC, an advanced microscopy laboratory established by IRCCS Ospedale San Raffaele and Università Vita-Salute San Raffaele. This work was supported by Fondazione Cariplo (E.C. and D.M.: 2014-1157).