Live sperm trap microarray for high throughput imaging and analysis
| Autor: | Yihe Wang, Thomas Hannam, David Sinton, Christopher McCallum, Jae Bem You, Alexander Lagunov, Farhang Tarlan, Keith Jarvi |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Male
medicine.medical_treatment Population Biomedical Engineering Motility Bioengineering Semen 02 engineering and technology Cell Separation Biology 01 natural sciences Biochemistry chemistry.chemical_compound Human fertilization Single-cell analysis medicine Humans Dimethylpolysiloxanes education education.field_of_study In vitro fertilisation 010401 analytical chemistry Acridine orange Optical Imaging General Chemistry DNA Microfluidic Analytical Techniques 021001 nanoscience & nanotechnology Sperm Spermatozoa 0104 chemical sciences Cell biology chemistry 0210 nano-technology |
| Zdroj: | Lab on a chip. 19(5) |
| ISSN: | 1473-0189 |
| Popis: | There is a growing appreciation and understanding of cell-to-cell variability in biological samples. However, research and clinical practice in male fertility has relied on population, or sample-based characteristics. Single-cell resolution is particularly important given the winner-takes-all nature of both natural and in vitro fertilization: it is the properties of a single cell, not the population, that are passed to the next generation. While there are a range of methods for single cell analysis, arraying a larger number of live sperm has not been possible due to the strong locomotion of the cells. Here we present a 103-trap microarray that traps, aligns and arrays individual live sperm. The method enables high-resolution imaging of the aligned cell head, the application of dye-based DNA and mitochondrial analyses, and the quantification of motility characteristics, such as tail beat. In testing, a 2400-post array trapped ∼400 sperm for individual analyses of tail beating frequency and amplitude, DNA integrity via acridine orange staining, and mitochondrial activity via staining. While literature results are mixed regarding a possible correlation between motility and DNA integrity of sperm at sample-level, results here find no statistical correlation between tail beat characteristics and DNA integrity at the cell-level. The trap array uniquely enables the high-throughput study of individual live sperm in semen samples - assessing the inherently single-cell selection process of fertilization, with single-cell resolution. |
| Databáze: | OpenAIRE |
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