Combination Usage of AdipoCount and Image-Pro Plus/ImageJ Software for Quantification of Adipocyte Sizes

Abstract

In recent decades, the prevalence of obesity has been rising. One of the major characteristics of obesity is fat accumulation, including hyperplasia (increase in number) and hypertrophy (increase in size). After histological staining, it is critical to accurately measure the number and size of adipocytes for assessing the severity of obesity in a timely fashion. Manual measurement is accurate but time-consuming. Although commercially available adipocyte counting tools, including AdipoCount, Image-Pro Plus, and ImageJ were helpful, limitations still exist in accuracy and time consuming. In the present study, we introduced the protocol of combined usage of these tools and illustrated the process with histological staining slides from adipose tissues of lean and obese mice. We found that the adipocyte sizes quantified by the tool combination were comparable as manual measurement, whereas the combined methods were more efficient. Besides, the recognition effect of monochrome segmentation image is better than that of color segmentation image. Overall, we developed a combination method to measure adipocyte sizes accurately and efficiently, which may be helpful for experimental process in laboratory and also for clinic diagnosis.

Liraglutide Activates Type 2 Deiodinase and Enhances β3-Adrenergic-Induced Thermogenesis in Mouse Adipose Tissue

“…Sections were imaged with a digital slide scanner (Leica Biosystems). ingWAT adipocyte diameter was determined using ImageJ, as previously described ( 14 ).…”

AdipoQ – a simple, open-source software to quantify adipocyte morphology and function in tissues and in vitro

“…Thus, we present a very straight-forward and fully automatized workflow that allows to analyze a dataset from raw data to plots in only three fully automated steps: two ImageJ plugins and automated post-hoc analysis with an R script. In contrast to previous approaches (Chen and Farese, 2002; Berry et al ., 2014; Parlee et al ., 2014; Maguire et al ., 2020; Hu et al ., 2021), AdipoQ, with its batch-processing modalities and easy accessibility through ImageJ and FIJI provides high-throughput analysis, freely available for all users. Furthermore, AdipoQ determines a broader set of parameters and also allows to combine read-outs from multiple fluorescence channels.…”

“…So far, automated image analysis to quantify adipogenesis in vitro only allowed to determine lipid droplet accumulation (Adomshick et al ., 2020). For analysing stained tissue sections, several approaches have been described that allow to determine adipocyte count and size distribution (Chen and Farese, 2002; Berry et al ., 2014; Parlee et al ., 2014; Maguire et al ., 2020; Hu et al ., 2021). Yet, these approaches are only partially automated (Chen and Farese, 2002; Parlee et al ., 2014; Maguire et al ., 2020; Hu et al ., 2021), rely on commercial software (Chen and Farese, 2002; Parlee et al ., 2014), and/or involve manual, user-biased threshold steps (Parlee et al ., 2014; Hu et al ., 2021), limiting throughput, reproducibility, and comparability.…”

“…For analysing stained tissue sections, several approaches have been described that allow to determine adipocyte count and size distribution (Chen and Farese, 2002; Berry et al ., 2014; Parlee et al ., 2014; Maguire et al ., 2020; Hu et al ., 2021). Yet, these approaches are only partially automated (Chen and Farese, 2002; Parlee et al ., 2014; Maguire et al ., 2020; Hu et al ., 2021), rely on commercial software (Chen and Farese, 2002; Parlee et al ., 2014), and/or involve manual, user-biased threshold steps (Parlee et al ., 2014; Hu et al ., 2021), limiting throughput, reproducibility, and comparability. Of note, one approach is no longer available (Berry et al ., 2014).…”

AdipoQ—a simple, open-source software to quantify adipocyte morphology and function in tissues and in vitro

“…The copyright holder for this preprint this version posted June 26, 2022. ; https://doi.org/10.1101/2022.06.04.494793 doi: bioRxiv preprint several approaches have been described that allow to determine adipocyte count and size distribution (Chen and Farese, 2002;Berry et al, 2014;Parlee et al, 2014;Maguire et al, 2020;Hu et al, 2021). Yet, these approaches are only partially automated (Chen and Farese, 2002;Parlee et al, 2014;Maguire et al, 2020;Hu et al, 2021), rely on commercial software (Chen and Farese, 2002;Parlee et al, 2014), and/or involve manual, user-biased threshold steps (Parlee et al, 2014;Hu et al, 2021), limiting throughput, reproducibility, and comparability.…”