Abstract

46th annual meeting of the Radiation Research Society Conference, Louisville, KY Optical imaging of living cells and intact tissues by non-linear, simultaneous multiphoton molecular excitation provides inherent three-dimensional diffraction-limited resolution while minimizing photodamage and peripheral photobleaching. The multiphoton laser scanning microscope raster-scans a focused, high repetition-rate femtosecond pulse train of infrared laser light across biological samples and efficiently detects the resulting multiphoton excited fluorescence. This development has recently allowed quantitative visualization of cellular processes and tissue architecture that had been problematic or impossible with conventional microscopies. The physical principles, aspects of instrumentation developments, and photophysical characterization of molecular fluorophores that have enabled multiphoton microscopy will be discussed. Examples such as neurotransmitter secretion, skin morphology, drug metabolism in the intact colon, and simultaneous metabolic and photosensitizer imaging in t umor spheroids demonstrate a range of applications pursued with this novel form of microscopy at the Developmental Resource for Biophysical Imaging and Opto-electronics (DRBIO) at Cornell. Supported at DRBIO by the NIH RR04224. M.G.N. is an NIH Postdoctoral Fellow supported by NRSA 1 F32 CA 72225-02.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR004224-12
Application #
6206233
Study Section
Project Start
1999-09-01
Project End
2000-08-31
Budget Start
1998-10-01
Budget End
1999-09-30
Support Year
12
Fiscal Year
1999
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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