Talk at DAMOP meeting, Santa Fe May 27-30 1998 Session I1 - Optical Techniques in Molecular Biology. NVITED session, Friday morning, May 29 FCS, a technique introduced in 1972 (Magde et al. 1972, Appl. Phys. Lett. 29:705), is currently being employed for ultrasensitive studies of molecular diffusion and aggregation, photophysical dynamics and fast chemical reactions. When performed in confocal geometries with one- or two-photon excitation (Denk et al. 1990, Science 248:73) of photostable dyes, its sensitivity enables us to follow the dynamics of sparse biologically relevant agents such as drugs and pathogens in solution as well as in living cells. Minimally interfering with biological functions of the observed system, FCS allows us to address a variety of important questions in molecular biology. We report that green fluorescent protein (GFP), a popular tool for in-vivo studies, is not only well suited for intracellular FCS, but moreover turned out to be a perfect candidate for the study of intramolecular dynamics. It exhibits a rapid flickering in the 100 ?s time range that we have identified to be due to reversible proto nation of the chromophore. Supported by NIH-P41.RR04224, NSF-D1R 8800278

National Institute of Health (NIH)
National Center for Research Resources (NCRR)
Biotechnology Resource Grants (P41)
Project #
Application #
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Cornell University
United States
Zip Code
Migone, Fernando F; Cowan, Robert G; Williams, Rebecca M et al. (2016) In vivo imaging reveals an essential role of vasoconstriction in rupture of the ovarian follicle at ovulation. Proc Natl Acad Sci U S A 113:2294-9
O'Dell, Ryan S; Cameron, David A; Zipfel, Warren R et al. (2015) Reelin Prevents Apical Neurite Retraction during Terminal Translocation and Dendrite Initiation. J Neurosci 35:10659-74
Byrnes, Laura J; Singh, Avtar; Szeto, Kylan et al. (2013) Structural basis for conformational switching and GTP loading of the large G protein atlastin. EMBO J 32:369-84
Jain, Manu; Robinson, Brian D; Scherr, Douglas S et al. (2012) Multiphoton microscopy in the evaluation of human bladder biopsies. Arch Pathol Lab Med 136:517-26
Degala, Satish; Williams, Rebecca; Zipfel, Warren et al. (2012) Calcium signaling in response to fluid flow by chondrocytes in 3D alginate culture. J Orthop Res 30:793-9
O'Dell, Ryan S; Ustine, Candida J M; Cameron, David A et al. (2012) Layer 6 cortical neurons require Reelin-Dab1 signaling for cellular orientation, Golgi deployment, and directed neurite growth into the marginal zone. Neural Dev 7:25
McMullen, J D; Kwan, A C; Williams, R M et al. (2011) Enhancing collection efficiency in large field of view multiphoton microscopy. J Microsc 241:119-24
Kim, Sally A; Sanabria, Hugo; Digman, Michelle A et al. (2010) Quantifying translational mobility in neurons: comparison between current optical techniques. J Neurosci 30:16409-16
Bowles, Robby D; Williams, Rebecca M; Zipfel, Warren R et al. (2010) Self-assembly of aligned tissue-engineered annulus fibrosus and intervertebral disc composite via collagen gel contraction. Tissue Eng Part A 16:1339-48
McMullen, Jesse D; Zipfel, Warren R (2010) A multiphoton objective design with incorporated beam splitter for enhanced fluorescence collection. Opt Express 18:5390-8

Showing the most recent 10 out of 68 publications