Quantitative Urine Sediment Scoring

Urine sediment scoring was performed as described previously(17 (link)). Mouse urine was diluted 1:10 and centrifuged onto glass slides using a Cytospin 3 (Thermo Shandon) at 1000 rpm for 3 minutes. Slides were covered with Wright-Giemsa stain for 30 seconds, washed twice with water, and visualize by light microscopy on Olympus BX41 brightfield microscope at 200X magnification (hpf). The average number of polymorphonuclear (PMN) cells was calculated from counting 2 independent fields and scored by a semi-quantitative scoring system of 0–5: 0, < 1 PMN/hpf; 1, 1–5 PMN/hpf; 2, 6–10 PMN/hpf; 3, 11–20 PMN/hpf, 4, 21–40 PMN/hpf, and 5, > 40 PMN/hpf.

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Mercado-Evans V., Chew C., Serchejian C., Saltzman A., Mejia M.E., Zulk J.J., Cornax I., Nizet V, & Patras K.A. (2024). Tamm-Horsfall protein augments neutrophil NETosis during urinary tract infection. bioRxiv.

Publication Preprint 2024

BX41 brightfield microscope

Corresponding Organization :

Other organizations : Baylor College of Medicine, University of California, San Diego, Institute of Metagenomics and Microbial Technologies (Slovenia)

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Variable analysis

independent variables

Urine dilution ratio (1:10)

dependent variables

Number of polymorphonuclear (PMN) cells in mouse urine

control variables

Centrifugation speed (1000 rpm)
Centrifugation duration (3 minutes)
Staining method (Wright-Giemsa stain for 30 seconds)
Microscope magnification (200X)

controls

Not explicitly mentioned
Not explicitly mentioned

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