Abstract

The equations expressing the intensity of illumination and the overall point-spread function (PSF) of a two-lens system, from Richards and Wolf (1959), were solved numerically, and the effect of a detector aperture was considered. The effects of the numerical aperture, of the radius of the detector aperture, of overfilling or underfilling the objective lens back-aperture, and of using one- or two-photon excitation, on the overall PSF and on the collection volume, were considered. Preliminary results show that the effect of underfilling the objective lens to 80% or higher has little effect on the profile or dimensions of the collection volume. These results are compared with calculations using the paraxial theory and pseudoparaxial theory to describe the light at the focus of the same lens system. These results do not explain the apparently large values of the structure factor (focal volume axial to radial aspect ratio) obtained in fluorescence correlation spectroscopic (FCS) measur ements in our laboratory (Petra Schwille, Sam Hess, unpublished results, 1998). B. Richards and E. Wolf, """"""""Electromagnetic diffraction in optical systems. II. Structure of the image field in an aplanatic system,"""""""" Proc. R. Soc. London Ser. A 253, 358-379 (1959).

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR004224-15
Application #
6657731
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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