Void spot assay visualization optimization for use of Void Whizzard in rats (Rattus norvegicus)

Abstract Void spot assay (VSA) noninvasively evaluates urination. This study optimizes VSA by comparing post‐assay paper visualization techniques: bright field light (BF), ultraviolet light (UV), and ninhydrin spray (N). Male rats were placed in filter paper lined cages for 4 h. After the assay, all...

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Main Authors: Hannah Ruetten, Jadyn Bothe, Shannon Lankford, Gopal Badlani, James Koudy Williams
Format: Article
Language:English
Published: Wiley 2025-05-01
Series:Physiological Reports
Subjects:
Online Access:https://doi.org/10.14814/phy2.70358
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author Hannah Ruetten
Jadyn Bothe
Shannon Lankford
Gopal Badlani
James Koudy Williams
author_facet Hannah Ruetten
Jadyn Bothe
Shannon Lankford
Gopal Badlani
James Koudy Williams
author_sort Hannah Ruetten
collection DOAJ
description Abstract Void spot assay (VSA) noninvasively evaluates urination. This study optimizes VSA by comparing post‐assay paper visualization techniques: bright field light (BF), ultraviolet light (UV), and ninhydrin spray (N). Male rats were placed in filter paper lined cages for 4 h. After the assay, all papers were dried. BF images were photographed (digital camera). UV images were captured using a Darkroom ultraviolet imaging cabinet. Papers were sprayed with ninhydrin and photographed (digital camera). All images were converted to binary for analysis with Void Whizzard. UV versus BF significantly differed in area. All three groups significantly differed in overall spot count and spots 0–0.1 cm2. UV versus N and UV versus BF significantly differed in 0.1–0.25 cm2 spots, UV versus N in 0.25–0.5 cm2, and N versus BF in spots 0.5–1 cm2. Overall BF visualization proved difficult. N provided an ideal way to highlight urine and image with a digital camera. Human fingerprints from pre‐assay handling of paper interfered with the analysis of the smallest sized spots; however, there were no differences in the detection of larger spots, spot distribution, or overall spot area. This study contributes to the development of a standardized VSA protocol for assessing bladder function in rodent models.
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spelling doaj-art-459e289c462b4bf9bb1e9b3e73d906572025-08-20T02:30:32ZengWileyPhysiological Reports2051-817X2025-05-01139n/an/a10.14814/phy2.70358Void spot assay visualization optimization for use of Void Whizzard in rats (Rattus norvegicus)Hannah Ruetten0Jadyn Bothe1Shannon Lankford2Gopal Badlani3James Koudy Williams4Wake Forest Institute for Regenerative Medicine Wake Forest Baptist Medical Center Winston‐Salem North Carolina USAWake Forest Institute for Regenerative Medicine Wake Forest Baptist Medical Center Winston‐Salem North Carolina USAWake Forest Institute for Regenerative Medicine Wake Forest Baptist Medical Center Winston‐Salem North Carolina USADepartment of Urology Wake Forest Baptist Medical Center Winston‐Salem Winston‐Salem North Carolina USAWake Forest Institute for Regenerative Medicine Wake Forest Baptist Medical Center Winston‐Salem North Carolina USAAbstract Void spot assay (VSA) noninvasively evaluates urination. This study optimizes VSA by comparing post‐assay paper visualization techniques: bright field light (BF), ultraviolet light (UV), and ninhydrin spray (N). Male rats were placed in filter paper lined cages for 4 h. After the assay, all papers were dried. BF images were photographed (digital camera). UV images were captured using a Darkroom ultraviolet imaging cabinet. Papers were sprayed with ninhydrin and photographed (digital camera). All images were converted to binary for analysis with Void Whizzard. UV versus BF significantly differed in area. All three groups significantly differed in overall spot count and spots 0–0.1 cm2. UV versus N and UV versus BF significantly differed in 0.1–0.25 cm2 spots, UV versus N in 0.25–0.5 cm2, and N versus BF in spots 0.5–1 cm2. Overall BF visualization proved difficult. N provided an ideal way to highlight urine and image with a digital camera. Human fingerprints from pre‐assay handling of paper interfered with the analysis of the smallest sized spots; however, there were no differences in the detection of larger spots, spot distribution, or overall spot area. This study contributes to the development of a standardized VSA protocol for assessing bladder function in rodent models.https://doi.org/10.14814/phy2.70358lower urinary tract dysfunctionmicturitionninhydrinurinary dysfunctionurine
spellingShingle Hannah Ruetten
Jadyn Bothe
Shannon Lankford
Gopal Badlani
James Koudy Williams
Void spot assay visualization optimization for use of Void Whizzard in rats (Rattus norvegicus)
Physiological Reports
lower urinary tract dysfunction
micturition
ninhydrin
urinary dysfunction
urine
title Void spot assay visualization optimization for use of Void Whizzard in rats (Rattus norvegicus)
title_full Void spot assay visualization optimization for use of Void Whizzard in rats (Rattus norvegicus)
title_fullStr Void spot assay visualization optimization for use of Void Whizzard in rats (Rattus norvegicus)
title_full_unstemmed Void spot assay visualization optimization for use of Void Whizzard in rats (Rattus norvegicus)
title_short Void spot assay visualization optimization for use of Void Whizzard in rats (Rattus norvegicus)
title_sort void spot assay visualization optimization for use of void whizzard in rats rattus norvegicus
topic lower urinary tract dysfunction
micturition
ninhydrin
urinary dysfunction
urine
url https://doi.org/10.14814/phy2.70358
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