Microfluidic Sperm Sorting Chips Providing Improved Outcomes for ART Procedures
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1 Microfluidic Sperm Sorting Chips Providing Improved Outcomes for ART Procedures
2 FERTILE and FERTILE PLUS TM FERTILE TM FERTILE PLUS FERTILE and FERTILE PLUS are flow-free sterile single-use chips for the sorting and separation of healthy normal sperm from the many compromised sperm present in a semen sample. FERTILE is designed to sort sperm for subsequent use in ICSI (intracytoplasmic sperm injection). FERTILE PLUS is designed to sort sperm for subsequent use in ICSI, IVF (in-vitro fertilization) and IUI (intrauterine insemination). FERTILE and FERTILE PLUS eliminate the use of sperm-damaging procedures such as centrifugation and mixing.
3 FERTILE A schematic of space-constrained microfluidic sorting (SCMS) system. (A) The SCMS microchip has different channel lengths for effective sperm sorting. (B) A microscope image of the inlet under a 2X objective. (C) A microscope image of a microchannel under a 10X objective. (D) A microscope image of the outlet under a 2X objective. Scale bars for the channel inlets and outlets are 1 cm. (E) Sperm tracking with the aid of ImageJ (NIH). (F) A schematic of the trajectory of a sperm (Reproduced from Ref. Tasoglu et al with permission). (G) FERTILE-ICSI product. The FERTILE chip is a flow-free single-use device with five lanes each consisting of an inlet port connected to a larger outlet collection port by a microfluidic channel 15mm L x 4mm W x 50µ D. In operation, each channel is filled with 13µl of sorting solution through the inlet port followed by 2µl of untreated semen. The chip is then incubated for 30 minutes. The limited depth of the channel constrains the migration of compromised sperm while allowing the most motile healthy sperm to migrate to the outlet where they are available for harvest.
4 FERTILE PLUS The FERTILE PLUS is a flow-free dual chambered single-use device. The first chamber contains a sample inlet and fluid channels separated from the second collection chamber by a microporous membrane. Channel dimensions and membrane porosity are designed to optimize the sorting and collection of the most motile sperm. Sorting is accomplished by the passage of sperm through the micropores of the membrane. The FERTILE PLUS chip has a single channel accommodating a semen sample size of 850µl.
5 FERTILE and FERTILE PLUS Features and Benefits Both chips eliminate sperm-damaging procedures associated with sperm washing, swim-up and gradient centrifugation. Sorted sperm exhibit better morphology, lower levels of reactive oxygen species (ROS) and less DNA fragmentation than the original semen sample. Pretreatment of the semen sample is not required; thereby, reducing the risk of contamination. The chips are user-friendly. They provide excellent sorting and yield within 30 minutes and eliminate the prep times inherent to other methods. FERTILE and FERTILE PLUS are simple to use platforms. They do not require extensive user training. Applications Assisted Reproductive Technology (ART) laboratories Andrology laboratories Embryology laboratories Veterinary ART
6 MOTILITY Percent Motility - FERTILE: There is a significant difference in motility between the non-sorted semen, swim-up and the sorted sperm indicative of the sorting capability of the FERTILE.(1,2) Curvilinear Velocity - FERTILE: After sorting with FERTILE, collected sperm have more than 1.5 times the curvilinear velocity compared to swim-up and non-sorted semen.(1,2) Straight-line Velocity - FERTILE: After sorting with FERTILE, collected sperm have more than 3.8 times the straight-line velocity compared to swim-up and non-sorted semen.(1,2)
7 MORPHOLOGY Microscopic Morphology Analysis - FERTILE PLUS: Sorting with FERTILE PLUS results in an almost 2-fold increase in normal morphology compared to non-sorted semen.(2)
8 REACTIVE OXYGEN SPECIES Reactive Oxygen Species (ROS) Analysis - FERTILE PLUS: Sorting with FERTILE PLUS results in an approximately 5-fold reduction in ROS generation.(3,4,5) DNA FRAGMENTATION DNA Fragmentation Analysis - FERTILE PLUS: Sorting with FERTILE PLUS results in an approximately 20-fold reduction in DNA fragmentation compared to non-sorted semen.(3,4)
9 CERTIFICATES FERTILE and FERTILE PLUS are manufactured by KOEK Biotechnology under the terms of a sublicense with DxNow. KOEK Certifications: TSE EN ISO 9001:2008 QUALITY MANAGEMENT SYSTEM CERTIFICATION TSE EN ISO 13485:2012 MEDICAL DEVICES - QUALITY MANAGEMENT SYSTEM CERTIFICATION PATENTS FERTILETM and FERTILE PLUSTM are trademarks of DxNow, Inc. and KOEK Biotechnology. Patents Pending. Publication numbers: WO A2 and WO A1. REFERENCES 1. Tasoglu, S., Safaee, H., Zhang, X., Kingsley, J. L., Catalano, P. N., Gurkan, U. A., Nureddin, A., Kayaalp, E., Anchan, R. M., Maas, R. L., Tuzel, E., and Demirci, U. Exhaustion of racing sperm in nature-mimicking microfluidic channels during sorting. Small. 2013; 9(20): Waseem A., Velasco, V., Kingsley, J. L., Shoukat, M. S., Shafiee, H., Anchan, R. M., Mutter, G. L., Tuzel, E., and Demirci, U. Selection of functional sperm with higher DNA integrity and fewer reactive oxygen species. Advanced Healthcare Materials. 2014; 3(10): Aitken, R. J. and Clarkson, J.S., 1988, Significance of reactive oxygen species and antioxidants in defining the efficacy of sperm preparation techniques, Journal of Andrology. 1988; 9(6): Zini A., Finelli A., Phang D., Jarvi K. Influence of semen processing technique on human sperm DNA integrity, Urology. 2000; 56: Aitken, R. J., Bronson, R., Smith, T.B., and De Luliis, G.N. The source and significance of DNA damage in human spermatozoa; a commentary on diagnostic strategies and straw man fallacies. Molecular Human Reproduction. 2013; 19(8): Brown, D. B., Merryman, D. C., Rivnay, B., Houserman, V. L., Long, C. A., Honea, K. L. Evaluating a novel panel of sperm function tests for utility in predicting intracytoplasmic sperm injection (ICSI) outcome. Journal of Assisted Reproduction and Genetics. 2013; 30(4):
10 DxNow, Inc. One Innovation Drive Worcester, MA (Office)
Microfluidic Sperm Sorting Chips Providing Improved Outcomes for ART Procedures
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