Addition of formaldehyde releaser imidazolidinyl urea and MOPS buffer to urine samples enables delayed processing for flow cytometric analysis of urinary cells: A simple, two step conservation method of urinary cells for flow cytometry

IF 2.3 3区医学 Q3 MEDICAL LABORATORY TECHNOLOGY Cytometry Part B: Clinical Cytometry Pub Date : 2023-03-07 DOI:10.1002/cyto.b.22117

Paul Freund, Christopher M. Skopnik, Diana Metzke, Nina Goerlich, Jan Klocke, Emil Grothgar, Luka Prskalo, Falk Hiepe, Philipp Enghard

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Abstract

Introduction

Kidney diseases are a major health concern worldwide. Currently there is a large unmet need for novel biomarkers to non-invasively diagnose and monitor kidney diseases. Urinary cells are promising biomarkers and their analysis by flow cytometry has demonstrated its utility in diverse clinical settings. However, up to date this methodology depends on fresh samples, as cellular event counts and the signal-to-noise-ratio deter over time. Here we developed an easy-to-use two-step preservation method for conservation of urine samples for subsequent flow cytometry.

Methods

The protocol utilizes a combination of the formaldehyde releasing agent imidazolidinyl urea (IU) and MOPS buffer, leading to gentle fixation of urinary cells.

Results

The preservation method increases acceptable storing time of urine samples from several hours to up to 6 days. Cellular event counts and staining properties of cells remain comparable to fresh untreated samples.

Outlook

The hereby presented preservation method facilitates future investigations on flow cytometry of urinary cells as potential biomarkers and may enable broad implementation in clinical practice.

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在尿液样本中加入甲醛释放剂咪唑烷基脲和 MOPS 缓冲液，可延迟尿液细胞流式细胞术分析的处理时间：用于流式细胞仪的尿液细胞两步简单保存法

导言肾脏疾病是全球关注的主要健康问题。目前，对新型生物标记物的需求仍未得到满足，而这些标记物可用于无创诊断和监测肾脏疾病。尿液细胞是一种很有前景的生物标记物，流式细胞仪对其进行的分析已在不同的临床环境中证明了其实用性。然而，迄今为止，这种方法依赖于新鲜样本，因为细胞事件计数和信噪比会随着时间的推移而降低。在此，我们开发了一种简单易用的两步保存法，用于保存尿液样本，以便随后进行流式细胞术分析。方法该方法结合使用甲醛释放剂咪唑烷基脲（IU）和 MOPS 缓冲液，温和地固定尿液细胞。结果该保存方法可将尿液样本的可接受保存时间从数小时延长至 6 天。细胞事件计数和细胞染色特性仍与未经处理的新鲜样本相当。展望本文介绍的保存方法有助于今后对尿液细胞作为潜在生物标记物进行流式细胞术研究，并可在临床实践中广泛应用。

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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来源期刊

Cytometry Part B: Clinical Cytometry 医学-病理学

CiteScore

6.80

自引率

32.40%

发文量

审稿时长

>12 weeks

期刊介绍： Cytometry Part B: Clinical Cytometry features original research reports, in-depth reviews and special issues that directly relate to and palpably impact clinical flow, mass and image-based cytometry. These may include clinical and translational investigations important in the diagnostic, prognostic and therapeutic management of patients. Thus, we welcome research papers from various disciplines related [but not limited to] hematopathologists, hematologists, immunologists and cell biologists with clinically relevant and innovative studies investigating individual-cell analytics and/or separations. In addition to the types of papers indicated above, we also welcome Letters to the Editor, describing case reports or important medical or technical topics relevant to our readership without the length and depth of a full original report.