Functional studies of the kidney of living animals using multicolor two-photon microscopy.
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.
Dunn
Kenneth W
KW
Department of Medicine, Division of Nephrology, Indiana Center for Biological Microscopy, Indiana University Medical Center, Indianapolis 46202, USA. kwdunn@iupui.edu
Sandoval
Ruben M
RM
Kelly
Katherine J
KJ
Dagher
Pierre C
PC
Tanner
George A
GA
Atkinson
Simon J
SJ
Bacallao
Robert L
RL
Molitoris
Bruce A
BA
D000818:Animals; D017209:Apoptosis; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D004396:Coloring Agents / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004705:Endocytosis; D005456:Fluorescent Dyes / Q000008:administration & dosage / Q000493:pharmacokinetics; D005839:Gentamicins / Q000378:metabolism / Q000493:pharmacokinetics; D007275:Injections, Intravenous; D020407:Internet; D007668:Kidney / Q000166:cytology* / Q000473:pathology / Q000502:physiology*; D007687:Kidney Tubules, Proximal / Q000166:cytology; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008845:Microinjections; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D009336:Necrosis; D007690:Polycystic Kidney Diseases / Q000473:pathology; D051381:Rats; D011919:Rats, Inbred Strains; D012212:Rheology / Q000379:methods; D014018:Tissue Distribution; D014741:Video Recording
D004396:Coloring Agents; D005456:Fluorescent Dyes; D005839:Gentamicins; D004247:DNA