Trap Sperm
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Trap Sperm
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1 InnoGenomics Technologies, LLC. 2000 Lakeshore Dr #5016, New Orleans, LA 70148, USA. Electronic address: ssinha@innogenomics.com.
2 InnoGenomics Technologies, LLC. 2000 Lakeshore Dr #5016, New Orleans, LA 70148, USA.
Sudhir K Sinha et al.
Forensic Sci Int Genet .
2022 Jul .
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1 InnoGenomics Technologies, LLC. 2000 Lakeshore Dr #5016, New Orleans, LA 70148, USA. Electronic address: ssinha@innogenomics.com.
2 InnoGenomics Technologies, LLC. 2000 Lakeshore Dr #5016, New Orleans, LA 70148, USA.
White TJ, Rye MS, Tay JW.
White TJ, et al.
J Forensic Sci. 2022 Jul;67(4):1679-1689. doi: 10.1111/1556-4029.15043. Epub 2022 Apr 4.
J Forensic Sci. 2022.
PMID: 35373351
Alderson G, Gurevitch H, Casimiro T, Reid B, Millman J.
Alderson G, et al.
Forensic Sci Int Genet. 2018 Sep;36:225-232. doi: 10.1016/j.fsigen.2018.06.014. Epub 2018 Jul 9.
Forensic Sci Int Genet. 2018.
PMID: 30077873
Timken MD, Klein SB, Kubala S, Scharnhorst G, Buoncristiani MR, Miller KWP.
Timken MD, et al.
Forensic Sci Int Genet. 2019 May;40:96-104. doi: 10.1016/j.fsigen.2019.02.011. Epub 2019 Feb 13.
Forensic Sci Int Genet. 2019.
PMID: 30785062
Grosjean F, Favre M, Castella V.
Grosjean F, et al.
Int J Legal Med. 2022 Jun 30. doi: 10.1007/s00414-022-02861-7. Online ahead of print.
Int J Legal Med. 2022.
PMID: 35773355
Katilius E, Carmel AB, Koss H, O'Connell D, Smith BC, Sanders GM, LaBerge GS.
Katilius E, et al.
Forensic Sci Int Genet. 2018 Jul;35:9-13. doi: 10.1016/j.fsigen.2018.03.011. Epub 2018 Mar 27.
Forensic Sci Int Genet. 2018.
PMID: 29609058
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The Sperm X method uses a nanotechnology derived polymer membrane that functions as a separation medium to effectively trap sperm cells while enabling efficient flow through of the digested epithelial cell DNA. This specialized membrane enabled development of a method that could significantly increase a forensic laboratory's ability to obtain male sperm fraction DNA profiles. The SpermX device provides a rapid, reproducible procedure that is easy to implement in a single-tube format as well as high-throughput truly automated hands-free workflows. Validation studies, performed using the manual SpermX method, include sensitivity, stability, precision (reproducibility and repeatability), mixtures, and a method comparison to the traditional differential extraction. Sensitivity and method comparison studies demonstrated a wide range of sperm cells, from a high of over 2.78 million cells (9158 ng) to a low of 25 cells (83 pg), can be trapped by the SpermX membrane. Stability studies on various substrates (i.e., carpet, cotton, denim, polyester, and silk) and degraded semen gave the expected male DNA profiles. Data from the same operator and a different operator were consistent with low variance. Mixtures, with ratios ranging from approximately 10:1-18182:1, created to simulate real casework type samples including buccal/semen, vaginal epithelial/semen, and post coital swabs at different time intervals, were tested. A comparison of the SpermX method to the conventional differential extraction method resulted in comparable probative male profile allelic data and associated statistical probabilities. For low level sperm samples, down to 25 sperm cells (83 pg), the SpermX method outperformed the conventional differential extraction with more genotypic information and associated probabilities.
Keywords:
Differential extraction; Nanofibers; SpermX.
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Clipboard, Search History, and several other advanced features are temporarily unavailable.
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Affiliation
1 Department of Mechanical and Industrial Engineering, University of Toronto, 5 King's College Road, Toronto, ON M5S 3G8, Canada. sinton@mie.utoronto.ca.
Jae Bem You et al.
Lab Chip .
2019 .
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1 Department of Mechanical and Industrial Engineering, University of Toronto, 5 King's College Road, Toronto, ON M5S 3G8, Canada. sinton@mie.utoronto.ca.
Nagata MPB, Endo K, Ogata K, Yamanaka K, Egashira J, Katafuchi N, Yamanouchi T, Matsuda H, Goto Y, Sakatani M, Hojo T, Nishizono H, Yotsushima K, Takenouchi N, Hashiyada Y, Yamashita K.
Nagata MPB, et al.
Proc Natl Acad Sci U S A. 2018 Apr 3;115(14):E3087-E3096. doi: 10.1073/pnas.1717974115. Epub 2018 Mar 19.
Proc Natl Acad Sci U S A. 2018.
PMID: 29555773
Free PMC article.
de Wagenaar B, Berendsen JT, Bomer JG, Olthuis W, van den Berg A, Segerink LI.
de Wagenaar B, et al.
Lab Chip. 2015 Mar 7;15(5):1294-301. doi: 10.1039/c4lc01425a.
Lab Chip. 2015.
PMID: 25578490
Shao B, Shi LZ, Nascimento JM, Botvinick EL, Ozkan M, Berns MW, Esener SC.
Shao B, et al.
Biomed Microdevices. 2007 Jun;9(3):361-9. doi: 10.1007/s10544-006-9041-3.
Biomed Microdevices. 2007.
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Shamsi MB, Imam SN, Dada R.
Shamsi MB, et al.
J Assist Reprod Genet. 2011 Nov;28(11):1073-85. doi: 10.1007/s10815-011-9631-8. Epub 2011 Sep 9.
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PMID: 21904910
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Parmegiani L, Cognigni GE, Filicori M.
Parmegiani L, et al.
Adv Exp Med Biol. 2014;791:151-72. doi: 10.1007/978-1-4614-7783-9_10.
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You JB, McCallum C, Wang Y, Riordon J, Nosrati R, Sinton D.
You JB, et al.
Nat Rev Urol. 2021 Jul;18(7):387-403. doi: 10.1038/s41585-021-00465-1. Epub 2021 May 17.
Nat Rev Urol. 2021.
PMID: 34002070
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Gallagher MT, Cupples G, Ooi EH, Kirkman-Brown JC, Smith DJ.
Gallagher MT, et al.
Hum Reprod. 2019 Jul 8;34(7):1173-1185. doi: 10.1093/humrep/dez056.
Hum Reprod. 2019.
PMID: 31170729
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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.
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