Abstract
Cellular heterogeneity is a major hindrance, leading to the misunderstanding of dynamic cell biology. However, single cell analysis (SCA) has been used as a practical means to overcome this drawback. Many contemporary methodologies are available for single cell analysis; among these, microfluidics is the most attractive and effective technology, due to its advantages of low-volume specimen consumption, label-free evaluation, and real-time monitoring, among others. In this paper, a conceptual application for microfluidic single cell analysis for veterinary research is presented. A microfluidic device is fabricated with an elastomer substrate, polydimethylsiloxane (PDMS), under standard soft lithography. The performance of the microdevice is high-throughput, sensitive, and user-friendly. A total of 53.1% of the triangular microwells were able to trap single canine cutaneous mast cell tumor (MCT) cells. Of these, 38.82% were single cell entrapments, while 14.34% were multiple cell entrapments. The ratio of single-to-multiple cell trapping was high, at 2.7:1. In addition, 80.5% of the trapped cells were viable, indicating that the system was non-lethal. OCT4A-immunofluorescence combined with the proposed system can assess OCT4A expression in trapped single cells more precisely than OCT4A-immunohistochemistry. Therefore, the results suggest that microfluidic single cell analysis could potentially reduce the impact of cellular heterogeneity.
Highlights
Normal and neoplastic cells are capable of varying their biology in different ways over time
Cell heterogeneity is a serious problem, which hampers our understanding of real cell biology
There are many methods and technologies which can be applied for single cell analysis, microfluidics seems to be the most outstanding
Summary
Normal and neoplastic cells are capable of varying their biology in different ways over time. This ability maintains their homeostasis, in order to cope with the dynamic changes of extracellular and intracellular microenvironments [1]. The distinguished biological setup of each cell, they are in the same tissue, leads to the biological instability generally referred to as cellular heterogeneity. This anisotropic feature is influenced by genetic and epigenetic elements [2], microenvironments, cell-to-cell communications, and/or cell-to-acellular component interactions [3]. The strongest signal frequently interferes with the weaker signals produced from the rare cell groups of interest (e.g., cancer stem cells, adult stem cells, and so on)
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