Improved Model of Fluorescence Recovery Expands the Application of Multiphoton Fluorescence Recovery after Photobleaching in Vivo

Sullivan, Kelley D.; Sipprell, William H.; Brown, Edward B.

doi:10.1016/j.bpj.2009.04.020

Cited by 14 publications

(48 citation statements)

References 43 publications

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“…Multiphoton fluorescence after photobleaching (MP-FRAP) of injected FITC-BSA was used to calculate the lymph viscosity and speed [46]. It was found that there was no difference in lymph viscosity between WT, expanding or collapsed PLN mice.…”

Section: Measurement Of Lymph Node Pressure Lymphatic Pumping Prementioning

confidence: 99%

The role of the lymphatic system in inflammatory-erosive arthritis

Bouta

et al. 2015

Seminars in Cell & Developmental Biology

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Rheumatoid arthritis (RA) is a prevalent inflammatory joint disease with enigmatic flares, which causes swelling, pain, and irreversible connective tissue damage. Recently, it has been demonstrated in murine models of RA that the popliteal lymph node (PLN) is a biomarker of arthritic flare, as it “expands” in size and contrast enhancement during a prolonged asymptomatic phase, prior to when it “collapses” with accelerated synovitis and joint erosion. This PLN collapse is associated with adjacent knee flare, decreases in PLN volume and contrast enhancement, lymphatic pulse and pumping pressure, and an increase in PLN pressure. Currently, it is known that PLN collapse is accompanied by a translocation of B cells from the follicles to the sinuses, effectively clogging the lymphatic sinuses of the PLN, and that B cell depletion therapy ameliorates arthritic flare by eliminating these B cells and restoring passive lymphatic flow from inflamed joints. Here we review the technological advances that have launched this area of research, describe future directions to help elucidate the potential mechanism of PLN collapse, and speculate on clinical translation towards new diagnostics and therapies for RA.

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Section: Measurement Of Lymph Node Pressure Lymphatic Pumping Prementioning

confidence: 99%

The role of the lymphatic system in inflammatory-erosive arthritis

Bouta

et al. 2015

Seminars in Cell & Developmental Biology

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“…Due to the nonlinear excitation in multiphoton-based photobleaching experiments, only the fluorophore within the focal spot is photobleached and the photobleaching profile yields a well-defined 3D geometry 6, 13, 16 . Moreover, due to the deeper penetration of multiphoton imaging 12 , multiphoton fluorescence recovery after photobleaching (MP-FRAP) techniques can be applied to thick samples with large light scattering 5, 26 . However, current 3D MP-FRAP models and techniques focus on the isotropic diffusion measurements due to the primary application involving the study of protein mobilities within intracellular structures.…”

Section: Introductionmentioning

confidence: 99%

Measurement of Three-Dimensional Anisotropic Diffusion by Multiphoton Fluorescence Recovery after Photobleaching

et al. 2013

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The multiphoton fluorescence recovery after photobleaching (MP-FRAP) technique has been developed to measure the three-dimensional (3D) solute diffusion within biological systems. However, current 3D MP-FRAP models are based on isotropic diffusion and spatial domain analysis. The 3D anisotropic diffusion and frequency domain analysis for MP-FRAP measurements are rarely studied. In this study, a new technique is demonstrated for the quantitative and non-destructive determination of 3D anisotropic solute diffusion tensors within biological fibrosis tissues by multiphoton photobleaching and spatial Fourier analysis (SFA). Compared to the spatial domain analysis based MP-FRAP techniques, this SFA-based method has the capability for determining the 3D anisotropic diffusion tensors as well as the flexibility for satisfying initial and boundary conditions. First, a close-form solution of the 3D anisotropic diffusion equation is derived by solely using SFA. Next, this new method is validated by computer-simulated MP-FRAP experiments with pre-defined 3D anisotropic diffusion tensors as well as experimental diffusion measurements of FITC-Dextran (FD) molecules in aqueous glycerol solutions. Finally, this MP-FRAP technique is applied to the measurement of 3D anisotropic diffusion tensors of FD molecules within porcine tendon tissues. This study provides a new tool for complete determination of 3D anisotropic solute diffusion tensor in biological tissues.

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“…In this case, the shape of the fluorescence recovery curve changes, and so must the mathematical model describing the recovery. By introducing a simple coordinate shift into the derivation of the equation immediately above, the new derivation yields (Sullivan et al 2009)…”

Section: Introductionmentioning

confidence: 99%

“…All other variables are defined as above. With this “diffusion-convection” model, accurate values for the diffusion coefficient can be obtained even under the influence of moderate flows, defined by a dimensionless speed parameter, v s = v(w r /8D) ≤ 3 (Fig.3) (Sullivan et al 2009). …”

Section: Introductionmentioning

confidence: 99%

Single- and Two-Photon Fluorescence Recovery after Photobleaching

Sullivan¹,

Majewska²,

Brown³

2015

Cold Spring Harb Protoc

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Fluorescence recovery after photobleaching (FRAP) is a microscopy technique for measuring the kinetics of fluorescently labeled molecules, and can be applied both in vitro and in vivo for two-and three-dimensional systems. This chapter discusses the three basic FRAP methods: traditional FRAP, multi-photon FRAP (MPFRAP), and FRAP with spatial Fourier analysis (SFA-FRAP). Each discussion is accompanied by a description of the appropriate mathematical analysis appropriate for situations in which the recovery kinetics are dictated by free diffusion. In some experiments, the recovery kinetics are dictated by the boundary conditions of the system, and FRAP is then used to quantify the connectivity of various compartments. Since the appropriate mathematical analysis is independent of the bleaching method, the analysis of compartmental connectivity is discussed last, in a separate section.

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Improved Model of Fluorescence Recovery Expands the Application of Multiphoton Fluorescence Recovery after Photobleaching in Vivo

Cited by 14 publications

References 43 publications

The role of the lymphatic system in inflammatory-erosive arthritis

The role of the lymphatic system in inflammatory-erosive arthritis

Measurement of Three-Dimensional Anisotropic Diffusion by Multiphoton Fluorescence Recovery after Photobleaching

Single- and Two-Photon Fluorescence Recovery after Photobleaching

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