High-throughput sperm assay using label-free microscopy: morphometric comparison between different sperm structures of boar and stallion spermatozoa.
Abstract: The capacity for microscopic evaluation of sperm is useful for assisted reproductive technologies (ART), because this can allow for specific selection of sperm cells for in vitro fertilization (IVF). The objective of this study was to analyze the same sperm samples using two high-resolution methods: spatial light interference microscopy (SLIM) and atomic force microscopy (AFM) to determine if with one method there was more timely and different information obtained than the other. To address this objective, there was evaluation of sperm populations from boars and stallions. To the best of our knowledge, this is the first reported comparison when using AFM and high-sensitivity interferometric microscopy (such as SLIM) to evaluate spermatozoa. Results indicate that with the use of SLIM microscopy there is similar nanoscale sensitivity as with use of AFM while there is approximately 1,000 times greater throughput with use of SLIM. With SLIM, there is also allowace for the measurement of the dry mass (non-aqueous content) of spermatozoa, which may be a new label-free marker for sperm viability. In the second part of this study, there was analysis of two sperm populations. There were interesting correlations between the different compartments of the sperm and the dry mass in both boars and stallions. Furthermore, there was a correlation between the dry mass of the sperm head and the length and width of the acrosome in both boars and stallions. This correlation is positive in boars while it is negative in stallions.
Copyright © 2020 Elsevier B.V. All rights reserved.
Publication Date: 2020-05-23 PubMed ID: 32828395PubMed Central: PMC8842552DOI: 10.1016/j.anireprosci.2020.106509Google Scholar: Lookup
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Summary
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The research focuses on using high-resolution microscopy methods, namely spatial light interference microscopy (SLIM) and atomic force microscopy (AFM), for sperm evaluation in boars and stallions. Results revealed that SLIM provided similar data to AFM but with a significantly faster data collection rate and a new potential measure for sperm viability.
Methods in Focus: SLIM and AFM
- The two main methodologies used in this study were spatial light interference microscopy (SLIM) and atomic force microscopy (AFM).
- SLIM is a type of interferometric microscopy that provides high-resolution, label-free imaging. It involves the use of light to measure the difference in optical path length in a sample.
- AFM, on the other hand, is a type of scanning probe microscopy that can provide a three-dimensional profile of a sample. Its high-resolution imaging capabilities make AFM suitable for investigations at the nanoscale level.
Objective and Procedure
- The study aimed to compare results obtained from SLIM and AFM when used for sperm evaluation.
- Samples were sourced from boars and stallions, and the same sample sets were analyzed using both microscopy technologies.
- The main objective was to find out if one method could deliver quicker and different information than the other.
Findings and Implications
- Results indicated that while both technologies were capable of nanoscale sensitivity, SLIM was significantly faster, offering a throughput approximately 1,000 times greater than AFM.
- Beyond its efficiency, SLIM also allowed the measurement of the dry mass (non-aqueous content) of spermatozoa. This could serve as a new indicator of sperm viability, a measurement that would be useful in assisted reproductive technologies (ART).
- In the study’s second part, an analysis of two different sperm populations revealed correlations between the various compartments of the spermatozoa and the dry mass in both species.
- In both boars and stallions, a correlation was observed between the dry mass of the sperm head and the size (length and width) of the acrosome, a part of the sperm cell that contains enzymes to help the sperm penetrate the egg. Interestingly, this correlation was positive in boars and negative in stallions, suggesting species-specific variations in sperm cell properties.
Significance
- This research provides valuable information for advancements in ART, in particular for selecting sperm cells for in vitro fertilization (IVF).
- Furthermore, the indication that SLIM can be a quicker, equally sensitive alternative to AFM for sperm evaluation opens up new possibilities for faster, more efficient sperm sample analyses in the future.
Cite This Article
APA
Rubessa M, Feugang JM, Kandel ME, Schreiber S, Hessee J, Salerno F, Meyers S, Chu I, Popescu G, Wheeler MB.
(2020).
High-throughput sperm assay using label-free microscopy: morphometric comparison between different sperm structures of boar and stallion spermatozoa.
Anim Reprod Sci, 219, 106509.
https://doi.org/10.1016/j.anireprosci.2020.106509 Publication
Researcher Affiliations
- Department of Animal Science, University of Illinois, Urbana-Champaign, IL 61801, USA; Carl R. Woese Institute for Genomic Biology, Univ. of Illinois at Urbana-Champaign, USA.
- Department of Animal and Dairy Sciences, Mississippi State University, Mississippi State, MS 39762, USA.
- Quantitative Light Imaging Laboratory, Department of Electrical and Computer Engineering, Beckman Institute of Advanced Science and Technology, USA.
- Department of Animal Science, University of Illinois, Urbana-Champaign, IL 61801, USA.
- Department of Animal Science, University of Illinois, Urbana-Champaign, IL 61801, USA.
- Department of Animal Science, University of Illinois, Urbana-Champaign, IL 61801, USA.
- Department of Animal Science, University of Illinois, Urbana-Champaign, IL 61801, USA.
- Institute for Imaging & Analytical Technologies, Mississippi State University, Mississippi State, MS 39762, USA.
- Department of Bioengineering, University of Illinois, Urbana, Illinois 61801, USA; Quantitative Light Imaging Laboratory, Department of Electrical and Computer Engineering, Beckman Institute of Advanced Science and Technology, USA.
- Department of Animal Science, University of Illinois, Urbana-Champaign, IL 61801, USA; Carl R. Woese Institute for Genomic Biology, Univ. of Illinois at Urbana-Champaign, USA; Department of Bioengineering, University of Illinois, Urbana, Illinois 61801, USA. Electronic address: mbwheele@illinois.edu.
MeSH Terms
- Animals
- Cell Shape
- Fertilization in Vitro / veterinary
- High-Throughput Screening Assays / methods
- High-Throughput Screening Assays / veterinary
- Horses
- Male
- Microscopy / methods
- Microscopy / veterinary
- Semen Analysis / methods
- Semen Analysis / veterinary
- Species Specificity
- Sperm Head / ultrastructure
- Spermatozoa / cytology
- Spermatozoa / ultrastructure
- Staining and Labeling / methods
- Staining and Labeling / veterinary
- Swine
Grant Funding
- R01 GM129709 / NIGMS NIH HHS
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