Optical microscopy, when applied to living animals, provides a powerful means of studying cell biology in the most physiologically relevant setting. The ability of two-photon microscopy to collect optical sections deep into biological tissues has opened up the field of intravital microscopy to high-resolution studies of the brain, lens, skin, and tumors. Here we present examples of the way in which two-photon microscopy can be applied to intravital studies of kidney physiology. Because the kidney is easily externalized without compromising its function, microscopy can be used to evaluate various aspects of renal function in vivo. These include cell vitality and apoptosis, fluid transport, receptor-mediated endocytosis, blood flow, and leukocyte trafficking. Efficient two-photon excitation of multiple fluorophores permits comparison of multiple probes and simultaneous characterization of multiple parameters and yields spectral information that is crucial to the interpretation of images containing uncharacterized autofluorescence. The studies described here demonstrate the way in which two-photon microscopy can provide a level of resolution previously unattainable in intravital microscopy, enabling kinetic analyses and physiological studies of the organs of living animals with subcellular resolution.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1152/ajpcell.00159.2002 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
A novel ,,-coordinated organofluoroboron probe for amyloid detection: insight from experiment and theory.
Chem Commun (Camb)
January 2025
Faculty of Chemistry, Wroclaw University of Science and Technology, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland.
This work presents the results of photophysical studies for a newly synthesized BF-based organoboron dye of D-A-D topology. The one- and two-photon properties of the dye are compared with the D-A parent compound and commercially available amyloid marker: methoxy-X04. We demonstrate that the new dye exhibits better optical properties upon binding to amyloids than methoxy-X04, including emission above 600 nm, higher values of 2PA cross section, broader excitation range and higher increment of emission intensity upon binding to amyloids.
View Article and Find Full Text PDFAPOE4 and sedentary lifestyle synergistically impair neurovascular function in the visual cortex of awake mice.
Commun Biol
January 2025
School of Psychology and Sussex Neuroscience, University of Sussex, Brighton, UK.
Reduced cerebral blood flow occurs early in the development of Alzheimer's disease (AD), but the factors producing this reduction are unknown. Here, we ask whether genetic and lifestyle risk factors for AD-the ε4 allele of the Apolipoprotein (APOE) gene, and physical activity-can together produce this reduction in cerebral blood flow which leads eventually to AD. Using in vivo two-photon microscopy and haemodynamic measures, we record neurovascular function from the visual cortex of physically active or sedentary mice expressing APOE3 and APOE4 in place of murine APOE.
View Article and Find Full Text PDFIn living organisms, the natural motion caused by heartbeat, breathing, or muscle movements leads to the deformation of tissue caused by translation and stretching of the tissue structure. This effect results in the displacement or deformation of the plane of observation for intravital microscopy and causes motion-induced aberrations of the resulting image data. This, in turn, places severe limitations on the time during which specific events can be observed in intravital imaging experiments.
View Article and Find Full Text PDFBiological applications using multiphoton microscopy increasingly seek a larger field of view while maintaining sufficient temporal sampling to observe dynamic biological processes. Multiphoton imaging also requires high numerical aperture microscope objectives to realize efficient non-linear excitation and collection of fluorescence. This combination of low-magnification and high-numerical aperture poses a challenge for system design.
View Article and Find Full Text PDFAdaptive optics (AO) improves the spatial resolution of microscopy by correcting optical aberrations. While its application has been well established in microscopy modalities utilizing a circular pupil, its adaptation to systems with non-circular pupils, such as Bessel-focus two-photon fluorescence microscopy (2PFM) with an annular pupil, remains relatively uncharted. Herein, we present a modal focal AO (MFAO) method for Bessel-focus 2PFM.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!