Minimizing the variables of voiding spot assay for comparison between laboratories

Area of Science:

• Urology
• Animal Models
• Physiology

Background:

• The voiding spot assay (VSA) is a common method for assessing mouse urinary function.
• VSA results are sensitive to housing and procedural variables, leading to data inconsistency across laboratories.
• Key variables include analytical software, housing, transportation, and time of day.

Purpose of the Study:

• To evaluate the comparability of VSA results across laboratories by minimizing key variables.
• To identify critical factors influencing VSA data consistency and reliability.
• To establish standardized protocols for inter-laboratory VSA data comparison.

Main Methods:

• Comparative analysis of VSA data using Fiji and MATLAB analytical software.
• Assessment of VSA outcomes with mice housed in different daily cages.
• Evaluation of VSA sensitivity to transportation and time of day (morning vs. afternoon).
• Performing VSA with identical parameters in two geographically distinct laboratories.

Main Results:

• Analytical tools Fiji and MATLAB show good agreement for VSA parameter quantification, particularly for primary voiding spot (PVS) parameters.
• Mice housed in different daily cages did not significantly alter voiding patterns in a standard VSA cage.
• Mice exhibited high sensitivity to transportation and time of day, impacting voiding patterns.
• Limited comparable VSA data, such as PVS volume, can be generated across laboratories under identical procedural parameters.

Conclusions:

• Standardization of VSA protocols, including acclimation periods after transportation, is essential for reliable and comparable data.
• Minimizing variables related to transportation and time of day is critical for consistent VSA results.
• Comparable VSA data can be achieved across laboratories by adhering to identical procedural parameters, particularly for metrics like PVS volume.