Abstract

A technique for the measurement of the two-photon uncaging action cross section of calcium cages has been developed in this laboratory $^{1,2}$. Using this technique, we report on the prognosis for effective two-photon excitation of a novel calcium cage, DMNPE-4 $^{3}$. We also describe calculations which predict the total amount of calcium ions uncaged and the temporal behavior of the liberated calcium distribution under given multiphoton photolysis conditions with a given cage. These calculations, combined with the two-photon uncaging action cross sections that we have measured for a variety of calcium cages enable the quantitative study of calcium dependent processes with high spatial resolution within cells or thick tissue samples. The ultimate intent of these studies is the application of these tools to the quantitative study of the spontaneous transient calcium concentration elevations (""""""""sparks"""""""") in developing skeletal muscle cells, probing the potentiation/inhibition relation ships between consecutive sparks in one localized region as well as between sparks in spatially separate regions. 1. Shear, J. et al. { em Biophys. J.}70(2) A211 (1996) 2. Brown et al, Manuscript in Preparation (1997) 3. Ellis-Davies, G.{ em Biophys. J.}72(2) A246 (1997)

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR004224-15
Application #
6657810
Study Section
Project Start
2002-09-01
Project End
2003-08-31
Budget Start
Budget End
Support Year
15
Fiscal Year
2002
Total Cost
Indirect Cost

  Institution

Name
Cornell University
Department
Type
DUNS #
City
Ithaca
State
NY
Country
United States
Zip Code
14850

  Publications

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

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