ImageJ-FIJI-Assisted Estimation of Intestinal Layers' Volume: Study in Jejunum-Ileum of Rats.

Q3 Medicine Journal of Microscopy and Ultrastructure Pub Date : 2022-12-01 eCollection Date: 2024-10-01 DOI:10.4103/jmau.jmau_53_22
Satrio Adi Wicaksono, Yustina Andwi Ari Sumiwi, Dewi Kartikawati Paramita, Rina Susilowati
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Abstract

Background: Studies in the digestive tract often required precision quantification of intestinal volume to observe the effect of certain intervention/condition. Application of stereological methods could bring unbiased and accurate results but commercially computer-assisted systems are not widely available. ImageJ-FIJI is an open source software, which could become an alternative choice in the stereological measurement process.

Aim: This study describes simple stereological quantification methods during volume estimation of jejunum-ileum intestinal layers of the rats using a light microscope and ImageJ-FIJI stereological tool.

Material and methods: Six 3-months old male Sprague-Dawley rats were terminated and jejunum-ileum was harvested after perfusion. After removal of intestinal luminal content, whole jejunum-ileum weight was measured. The organ was sampled as 6-10 slabs of 1 cm length in a systematic uniformed random sampling manner. Slabs were cut longitudinally at random angles before flattened and put on filter papers for subsequent tissue processing into 2-3 paraffin blocks. One section of 3 μm thick was sampled from each block, stained using toluidine blue and documented using a light microscope connected to a microstepper apparatus. The volume of the intestinal layers was estimated using a point-counting grid on Image J-FIJI software.

Result: We compared two sets of counting methods i.e. minimal counting (MC) and rigorous counting (RC) approaches that differ in their respective a/p value. Quantification using RC approach resulted in significantly higher estimated volume of tunica submucosa and tunica muscularis while having more preferable stereological accuracy parameters (CE<5% & CV<10%).

Conclusion: Although it required longer counting time, rigorous approaches resulted in higher accuracy while still within the range of rule of thumb criteria of 0.2 < CE2/CV2 < 0.5.

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image - fiji辅助估计大鼠空肠-回肠肠层体积的研究。
背景:消化道研究往往需要精确定量肠容积来观察某种干预/条件的效果。应用立体学方法可以带来公正和准确的结果,但商用计算机辅助系统并不广泛使用。imagj - fiji是一个开源软件,它可以成为立体测量过程中的另一种选择。目的:介绍利用光学显微镜和imagj - fiji立体工具对大鼠空肠-回肠肠层进行体积估算的简单立体定量方法。材料与方法:处死6只3月龄雄性sd大鼠,灌注后取空肠-回肠。去除肠管内容物后,测量全空肠-回肠重量。以系统均匀随机抽样的方式,将器官取样为6-10块长度为1cm的薄片。将板坯按任意角度纵向切割,然后压平,放在滤纸上进行后续组织处理,切成2-3块石蜡。每个块取3 μm厚的切片,用甲苯胺蓝染色,用光学显微镜连接微步进仪记录。使用Image J-FIJI软件上的点计数网格估计肠层的体积。结果:我们比较了两组计数方法,即最小计数(MC)和严格计数(RC)方法,它们各自的a/p值不同。使用RC方法定量可以显著提高粘膜下层和肌肉层的估计体积,同时具有更好的立体学精度参数(cec结论:虽然需要更长的计数时间,但严格的方法可以提高精度,同时仍在0.2 < CE2/CV2 < 0.5的经验法则标准范围内。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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CiteScore
1.90
自引率
0.00%
发文量
46
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