An Improved Ocular Impression Cytology Method: Quantitative Cell Transfer to Microscope Slides Using a Novel Polymer.
An Improved Ocular Impression Cytology Method: Quantitative Cell Transfer to Microscope Slides Using a Novel Polymer.
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DOI：
10.1080/02713683.2021.1951300
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发表时间：
2022-01
影响因子：
2
通讯作者：
Rigas B
中科院分区：
文献类型：
--
作者：
Master A;Huang W;Huang L;Honkanen R;Rigas B
建立一种更有效的印迹细胞学（IC）方法，用于将眼表细胞转移到玻璃显微镜载玻片上进行细胞化学、免疫细胞化学和免疫荧光研究。用混合纤维素酯膜将细胞从眼表面剥离，然后使用由I型胶原蛋白、聚乙烯亚胺和聚-L-赖氨酸（CPP）组成的新型三嵌段共聚物，并通过加热和冷却使细胞和载玻片交联，将细胞牢固地附着到载玻片上。在用丁醇/乙醇溶液软化膜后，将其完整地移除。细胞转移在约10-15分钟内完成，并准备染色。我们的细胞转移方法的效率进行了比较，目前的方法的基础上聚-L-赖氨酸和白蛋白糊。我们的方法确保了几乎完全的细胞转移。相比之下，兔结膜细胞转移到聚-L-赖氨酸覆盖的载玻片上要低37.5± 6.3%，转移到白蛋白糊覆盖的载玻片上要低62.5± 5.6%（平均值± SD）;兔杯状细胞的转移效率甚至更低。新方法也更有效地转移从IC获得的人口腔粘膜细胞。转移的细胞成功地用H&E、化学发光和免疫荧光剂染色。使用我们的方法，我们对眼表细胞进行S100 A4和ATF 4染色，这两种细胞在干眼病的病理生理学中发挥作用。我们在口腔粘膜细胞中获得了类似的结果，这表明我们的方法具有普遍性。我们提出了一个解释的强烈粘附的细胞的玻璃载玻片，这是基于它们与三嵌段共聚物的相互作用。我们开发了一种新的方法，使用一种新的共聚物，用于有效和快速地将IC获得的细胞转移到玻璃显微镜载玻片上。与现有的方法相比，我们的改进方法使IC强大和简单，并应提高其诊断率和临床适用性。
To develop a more efficient impression cytology (IC) method for the transfer of ocular surface cells onto glass microscope slides for cytochemical, immunocytochemical and immunofluorescence studies. Cells are lifted off the ocular surface with a mixed cellulose ester membrane and then firmly attached to a glass slide using a novel triblock copolymer comprised of collagen type I, polyethylenimine and poly-L-lysine (CPP), and crosslinking cells and glass slide by heating and cooling. The membrane is removed intact after softening it with a butanol/ethanol solution. Transfer of cells is complete in about 10–15 minutes and are ready for staining. The efficiency of our cell transfer method was compared to current methods based on poly-L-lysine and albumin paste. Our method ensured almost complete transfer of cells. In contrast, the transfer of rabbit conjunctiva cells onto poly-L-lysine-covered slides was 37.5±6.3 % lower, and onto albumin-paste covered slides 62.5±5.6 % lower (mean ± SD); the transfer of rabbit goblet cells was even less efficient. The new method was also more efficient for transfer of cells from human oral mucosa obtained by IC. Transferred cells were successfully stained with H&E, chemiluminescence, and immunofluorescence agents. Using our method, we stained ocular surface cells for S100A4 and ATF4, both of which play a role in the pathophysiology of dry eye disease. We obtained similar results with oral mucosal cells, suggesting the generalizability of our approach. We propose an explanation for the strong adhesion of cells to the glass slide, which is based on their interactions with the triblock copolymer. We developed a novel approach for the efficient and rapid transfer of cells obtained by IC onto glass microscope slides using a novel copolymer. Compared to available methods, our improved approach makes IC robust and simple, and should increase its diagnostic yield and clinical applicability.
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影响因子：
7.4
作者：
Heilkenbrinker, Alexandra;Reinemann, Christine;Scheper, Thomas
通讯作者：
Scheper, Thomas
DOI：
10.1007/978-1-4939-2718-0_36
发表时间：
2015
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Kurien BT;Scofield RH
通讯作者：
Scofield RH
影响因子：
4.2
作者：
EGBERT, PR;LAUBER, S;MAURICE, DM
通讯作者：
MAURICE, DM
影响因子：
--
作者：
NELSON, JD;HAVENER, VR;CAMERON, JD
通讯作者：
CAMERON, JD
影响因子：
4.3
作者：
Holstein, Carly A.;Chevalier, Aaron;Yager, Paul
通讯作者：
Yager, Paul