Micro-sieving: isolation of whole glomeruli from a single renal needle biopsy sample.
Renal biopsy samples are important not only for the diagnosis of glomerulonephritis, but also for the investigation of its pathogenesis. However, it remains difficult to biochemically analyze proteins extracted solely from the glomeruli of needle biopsy samples, since the samples contain various components like renal tubules and connective tissue. Even a recent micro-dissection method, recovering the glomeruli in the sliced sections of the biopsy samples, has not fully solved the difficulty because the amount of obtainable proteins by this method is not usually enough for protein analysis. To overcome this problem, we established a simple but reliable method to isolate whole glomeruli from needle biopsy samples. By this method, termed 'micro-sieving', we were able to isolate on average more than 50 glomeruli from a single needle biopsy sample in an hour. The amount of the extracted glomerular proteins was on average 23 μg per biopsy sample. As a representative use of this method, we were able to obtain a glomerular protein profile by fluorescent 2-dimensional electrophoresis for each of the tested patients with glomerulonephritis. 'Micro-sieving' can be used widely as a fundamental technique to analyze glomeruli in renal needle biopsy samples.
Koitabashi
Kenichiro
K
Clinical Proteomics and Molecular Medicine, St. Marianna University Graduated School of Medicine, Kawasaki, Kanagawa, Japan.
Okamoto
Kazuki
K
Arirto
Mitsumi
M
Sato
Toshiyuki
T
Nagai
Kouhei
K
Kurokawa
Manae S
MS
Suematsu
Naoya
N
Yasuda
Takashi
T
Kimura
Kenjiro
K
Kato
Tomohiro
T
D001707:Biopsy, Needle / Q000295:instrumentation* / Q000379:methods*; D005921:Glomerulonephritis / Q000175:diagnosis / Q000473:pathology*; D006801:Humans; D007674:Kidney Diseases / Q000175:diagnosis / Q000473:pathology; D007678:Kidney Glomerulus / Q000473:pathology*