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Multiphoton imaging of sub‐podocyte space in isolated perfused glomeruli
Author(s) -
Salmon Andrew,
Toma Ildiko,
Harper Steven,
Bates David,
Neal Christopher,
PetiPeterdi Janos
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a503-a
Subject(s) - podocyte , pipette , afferent arterioles , fluorescence , fluorescence microscope , microscopy , biophysics , chemistry , anatomy , optics , biology , kidney , physics , endocrinology , biochemistry , angiotensin ii , receptor , proteinuria
Sub‐podocyte spaces (SPS), regions bounded by podocyte cell bodies or primary processes and glomerular capillary wall (GCW), cover 50–65% of the outer GCW aspect. The abundance and restrictive dimensions of SPS indicate a potential role in glomerular permeability a . We sought to identify SPS using multiphoton microscopy in living intact isolated perfused glomeruli. Single glomeruli were manually dissected from fresh cortical slices of rabbit kidney, and perfused through the afferent arteriole with 20μ m quinacrine via concentric pipettes. Tissue was excited with 860nm wavelength laser, and z‐stack images at 0.375μm intervals were recorded with Leica confocal software. SPS, identified as non‐fluorescent regions bounded by podocyte cell bodies/processes and GCW, had mean dimensions 2.3μm depth × 4.0μm width × 0.8μm height (n=5). Mean SPS height compares favourably with that reported using electron microscopy (0.3–1.3μm) a . Fluid and solute movement through SPS regions were characterised with different molecular weight fluorescent tracers. This technique confirms the presence of SPS in living tissue. Funding: Wellcome Trust 069134