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Full-Text Articles in Life Sciences

An Intersection Of Science & Art: Vitrification Approaches And Open-Fabricated Tools For The Biomedical Model Sea Hare, Aplysia Californica, Allyssa M. Oune Nov 2023

An Intersection Of Science & Art: Vitrification Approaches And Open-Fabricated Tools For The Biomedical Model Sea Hare, Aplysia Californica, Allyssa M. Oune

LSU Master's Theses

The California sea hare (Aplysia californica) is an important biomedical model for molecular neurobiology, electrophysiology, learning, and memory due to their well-mapped and large neurons and well-characterized learning capabilities. The National Resource for Aplysia (NRA, University of Miami) maintains large stocks of live animals and relies on regular shipments of wild-caught individuals to maintain genetic diversity. This is labor and cost-intensive, and environmental changes could alter the availability of wild animals increasing the need to preserve this genetic resource. One solution is vitrification, ultra-fast cooling which produces an amorphous glass that minimizes damage to cells. Aplysia californica presents …


A Modular Open-Technology Device To Measure And Adjust Concentration Of Sperm Samples For Cryopreservation, Nikolas C. Zuchowicz Sep 2021

A Modular Open-Technology Device To Measure And Adjust Concentration Of Sperm Samples For Cryopreservation, Nikolas C. Zuchowicz

LSU Master's Theses

Repositories for aquatic germplasm can safeguard the genetic diversity of species of interest to aquaculture, research, and conservation. The development of such repositories is impeded by a lack of standardization both within laboratories and across the research community. Protocols for cryopreservation are often developed ad hoc and without close attention to variables, such as sperm concentration, that strongly affect the success and consistency of cryopreservation. The wide dissemination and use of specialized tools and devices can improve processing reliability, provide data logging, produce custom hardware to address unique problems, and save costs, time, and labor. The goal of the present …


Vitrification Of Immature And Mature Bovine Oocytes, Paige T. Hardin Jan 2016

Vitrification Of Immature And Mature Bovine Oocytes, Paige T. Hardin

LSU Master's Theses

Vitrification is the latest technique used in cryopreservation, the ability to utilize this method with oocytes and embryos has become a valuable system. Vitrification has been successful with bovine embryos and oocytes but is far from optimal. Following cryopreservation storage discarding embryos can cause ethical issues, and mature oocytes have fragile organelles that can be detrimentally affected by ice crystal formation during freezing. Immature oocytes have not formed some of these temperature sensitive microfilaments and can circumvent these detrimental effects. The common intracellular cryoprotective agents are dimethyl sulfoxide, glycerol and ethylene glycol. Different combination of these agents have been reported …


Vitrification Of Equine Expanded Blastocysts, Fabian Andres Diaz Jan 2013

Vitrification Of Equine Expanded Blastocysts, Fabian Andres Diaz

LSU Master's Theses

The cryopreservation of equine expanded blastocysts (>300 um) has been largely unsuccessful primarily due to the low permeability to cryoprotectants and the large size of the equine embryo. Mechanical alternatives may provide means to overcome the capsule barrier and the relative large embryo size. In this regard, multiple experiments were performed in this study to evaluate different approaches of capsule puncture and blastocoele fluid extraction with the objective to develop a cryopreservation protocol for Day 8 equine expanded blastocysts. In the first experiment, twenty-four Day 8 expanded blastocysts were exposed to standard equine embryo vitrification solutions following one- or …


Development And Permeability Of Equine Blastocysts, Brittany Reshel Scott Jan 2011

Development And Permeability Of Equine Blastocysts, Brittany Reshel Scott

LSU Master's Theses

Equine embryo cryopreservation is unsuccessful in larger, more easily collected, day-7 embryos. It is imperative that methods to successfully cryopreserve large equine embryos or develop reliable methods to determine embryo size before collection. Therefore the objectives for this study were to quantify the amount of tritiated glycerol that would permeate various sizes of equine embryos and to determine if circulating progesterone concentration was correlated with in utero embryo size. Mean embryo diameter (± SEM) across treatments (1.4M and 3.4M tritiated glycerol) was 696.5µm ± 108.6µm and 925.9 µm ± 214.1µm, respectively and were not different (P=0.44). The percent permeation for …


Cryopreservation Of White-Tail Deer Epididymal Sperm For Artificial Insemination, Jesse Ray Saenz Jan 2007

Cryopreservation Of White-Tail Deer Epididymal Sperm For Artificial Insemination, Jesse Ray Saenz

LSU Master's Theses

The ability to cryopreserve epididymal sperm from mature postmortem bucks has long been of interest to both wildlife conservationists and deer ranchers. Increased understanding of the cryobiology of epididymal sperm from a non domestic species, such as White-tail deer, could aid in development of future protocols to assist in the preservation of endangered species. In Experiment I, results showed that after cooling postmortem bull testes for 22 hours, no significant difference was noted between sperm parameters of epididymal sperm collected at room temperature or at a cool enviorment. In Experiment II, it was shown that White-tail deer sperm could be …


Analysis Of U.S. Aquacultural Producer Preferences For Genetic Improvement And Cryopreservation, Brian Paul Boever Jan 2006

Analysis Of U.S. Aquacultural Producer Preferences For Genetic Improvement And Cryopreservation, Brian Paul Boever

LSU Master's Theses

Aquaculture industries in the U.S. generate $1 billion in farm-level sales. Genetic improvement of fish stocks may be a way to increase the market share of aquaculture within the U.S. seafood market as well as the market share within the world market. This study evaluates the preferences, beliefs, and opinions of aquaculture producers across the U.S. about topics such as cryopreservation, genetic improvement, and the future of the aquaculture industry. Willingness-to-pay values for specific genetic improvements by aquaculture grow-out producers were elicited. A national survey of aquaculture producers was used to elicit the information used in the analysis. The survey …


Cryopreservation Of Bovine And Caprine Oocytes By Vitrification, Sabrina Marie Luster Jan 2004

Cryopreservation Of Bovine And Caprine Oocytes By Vitrification, Sabrina Marie Luster

LSU Master's Theses

Cryopreservation of animal oocytes will permit germplasm of valuable or unique females to be preserved for extended times. The objective of this research was to derive a procedure to cryopreserve bovine oocytes by vitrification to be used as recipients for somatic cell nuclear transfer (SCNT). Caprine oocytes vitrified by the same procedure were assayed by cytological examination of microtubules. In the first two of three experiments, bovine oocytes matured in vitro were vitrified in a mixture of ethylene glycol (EG), dimethylsulfoxide (Me2SO) and trehalose, and then subjected to in vitro fertilization (IVF) or SCNT. For vitrification, oocytes were first exposed …


Survival Of Canine Epididymal Sperm Under Cooled And Frozen-Thawed Conditions, Karla Stilley Jan 2001

Survival Of Canine Epididymal Sperm Under Cooled And Frozen-Thawed Conditions, Karla Stilley

LSU Master's Theses

The objective of the first experiment was to determine the effects of storage at 22„aC vs. 4„aC on the motility and percentage of membrane-intact sperm (%MIS) of epididymal mouse sperm. Testicles were allocated into 22„aC or 4„aC treatment groups and stored for 24 or 48 hours. Additional testicles were allocated into the 4„aC treatment for storage for 72 or 96 hours. Sperm was collected and analyzed at each time point. Storage at 22„aC lowered motility and %MIS (P<0.05) when compared with control sperm and 4„aC sperm at both the 24 and 48 hours. Motility and %MIS of 4„aC sperm did not decrease when compared with the control until after 72 hours of storage. The second experiment evaluated the effects of 22„aC vs. 4„aC storage for 24 and 48 hours on epididymal dog sperm. Motility and %MIS of the 22„aC sperm was lower than that of the 4„aC sperm and the control (P<0.05) at 24 and 48 hours. Motility of the 4„aC sperm was lower than the control at 24 and 48 hours (P<0.05), however %MIS was not lower than the control until 48 hours. The third experiment tested the effect of cryopreservation on epididymal dog sperm. Sperm was frozen immediately (A), after 48 hours at 4„aC in liquid (B) or after 48 hours at 4„aC of the whole testicle (C). Both pre-freeze (PF) and post-thaw (PT) motility and %MIS of B and C were lower than A (P<0.05). PT values were lower than PF values in all treatments (P<0.05). PT motility of B was lower than C (0 vs. 35.0¡Ó3.6%). Storage at 4„aC allows collection of motile epididymal mouse and dog sperm for several days after death. Dog testicles can be refrigerated for 2 days and epididymal sperm frozen with PT motility and %MIS of 35 and 62%.