Ultra Rapid Laser Warming For Preservation of Fish and Other Aquatic Species

Khosla, Kanav; Smith, K D; Gangwar, Lakshya; Joshi, Amey; Liu, Yilin; Han, Guebum; McAlpine, Michael C.; Kodandaramiah, Suhasa; Hagedorn, Mary; Bischof, John C.

doi:10.1016/j.cryobiol.2021.11.062

Cited by 3 publications

(7 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Nevertheless, the successful vitrification achieved by CondD‐C allows us to further investigate complementary rewarming technologies, such as cryomesh Joule heating [ 35 ] or laser rewarming in a higher throughput configuration. [ 60 ]…”

Section: Resultsmentioning

confidence: 99%

“…The first successful protocol for zebrafish embryo vitrification employed a cryotop-like device made with a polypropylene strip. [58][59][60] However, such a protocol can vitrify one zebrafish embryo on cryo-top at a time, considerably limiting the throughput of cryopreservation. With the previous protocol, a well-trained operator can only vitrify 10-15 embryos in an hour.…”

Section: Zebrafish Embryo Cryopreservationmentioning

confidence: 99%

“…The high cooling rate of CondD-C demonstrated the highest viability improvements in small biosystems (i.e., coral larvae, Figure S20A, Supporting Information), but also demonstrated advantages for the vitrification of Drosophila embryos (Figure S20B, Supporting Information) and zebrafish embryos (Figure S20C, Supporting Information). With the increase of biosystem size (above ≈200 μm), the CondD-C has a good vitrification rate during cooling, but more rapid heating approaches [35,60] are required to further increase the viability during rewarming with low CPA concentration (below ≈20%).…”

Section: Design and Physical Limits Of The Cryomesh Platform Techniquementioning

confidence: 99%

See 2 more Smart Citations

Conduction‐Dominated Cryomesh for Organism Vitrification

Guo,

Zuchowicz,

Bouwmeester

et al. 2023

Advanced Science

Self Cite

View full text Add to dashboard Cite

Vitrification‐based cryopreservation is a promising approach to achieving long‐term storage of biological systems for maintaining biodiversity, healthcare, and sustainable food production. Using the “cryomesh” system achieves rapid cooling and rewarming of biomaterials, but further improvement in cooling rates is needed to increase biosystem viability and the ability to cryopreserve new biosystems. Improved cooling rates and viability are possible by enabling conductive cooling through cryomesh. Conduction‐dominated cryomesh improves cooling rates from twofold to tenfold (i.e., 0.24 to 1.2 × 105 °C min−1) in a variety of biosystems. Higher thermal conductivity, smaller mesh wire diameter and pore size, and minimizing the nitrogen vapor barrier (e.g., vertical plunging in liquid nitrogen) are key parameters to achieving improved vitrification. Conduction‐dominated cryomesh successfully vitrifies coral larvae, Drosophila embryos, and zebrafish embryos with improved outcomes. Not only a theoretical foundation for improved vitrification in µm to mm biosystems but also the capability to scale up for biorepositories and/or agricultural, aquaculture, or scientific use are demonstrated.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Zebrafish Embryo Cryopreservationmentioning

confidence: 99%

Section: Design and Physical Limits Of The Cryomesh Platform Techniquementioning

confidence: 99%

See 1 more Smart Citation

Conduction‐Dominated Cryomesh for Organism Vitrification

Guo,

Zuchowicz,

Bouwmeester

et al. 2023

Advanced Science

Self Cite

View full text Add to dashboard Cite

show abstract

“…Cryopreservation at scale could potentially reduce significant long-term costs and space needs by banking valuable zebrafish lines that are not routinely used. Recently, microinjection of a mixture of cryoprotective agents and gold nanorods have been explored for improving vitrification rates of whole intact zebrafish embryos followed by uniform rewarming of embryos using pulsed infrared lasers [11], [12], [38]. These methods have been successful in reanimating a small subset (3%) [12] of embryos which successfully survived to adulthood (Fig.…”

Section: High Throughput Cryopreservation Of Whole Intact Zebrafish E...mentioning

confidence: 99%

“…The cryopreservation and laser nanowarming technique used was similar to our previous study [11], [12], [38]. Zebrafish embryos were microinjected with 10 nl of CPA and GNRs (80% propylene glycol (PG) and 20% methanol (MeOH) with 100 µg/ml GNR) at the center of the yolk of zebrafish embryo at high cell stage (Fig.…”

Section: Cryopreservation and Laser Nanowarming Experimentsmentioning

confidence: 99%

A machine vision guided robot for fully automated embryonic microinjection

Alegria

Joshi

Mendana

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Microinjection is a widely used technique for transgenesis, mutagenesis, cell labeling, cryopreservation, and in vitro fertilization in multiple single and multicellular organisms. Microinjection requires specialized skills acquired for each target organism and involves rate limiting and labor-intensive preparatory steps. Here we constructed a machine vision (MV) guided generalized robot that fully automates the process of microinjection in fruit fly (Drosophila melanogaster) and zebrafish (Danio rerio) embryos. The robot uses machine learning (ML) models trained to detect individual embryos in images of agar plates, and models trained to identify specific anatomical locations within each embryo in 3D space using dual view microscopes. The robot uses this information to serially perform microinjection in each detected embryo without any human intervention. We constructed and used three such robots to automatically microinject tens of thousands of Drosophila and zebrafish embryos. We systematically optimized robotic microinjection for each species and validated the use of the robot by performing routine transgenesis with proficiency comparable to highly skilled human practitioners while achieving up to 4x increases in microinjection throughput in Drosophila. The automated microinjection robot was utilized to microinject pools of over 20,000 uniquely barcoded plasmids into 1,713 embryos in two days to rapidly generate more than 400 unique transgenic Drosophila lines. This high throughput microinjection experiment enabled a novel measurement of the number of independent germline integration events per successfully injected embryo. Finally, we showed that robotic microinjection of cryoprotective agents in zebrafish embryos significantly improves vitrification rates and survival of cryopreserved embryos post-thaw as compared to manual microinjection, opening the tantalizing possibility of large-scale cryobanking of aquatic species at an industrial scale. We anticipate that this versatile automated microinjection system can be applied to carry out microinjection for genome-wide manipulation and cryopreservation at scale in a wide range of other organisms.

show abstract

High-throughput genetic manipulation of multicellular organisms using a machine-vision guided embryonic microinjection robot

Alegria,

Joshi,

Mendana

et al. 2024

Genetics

Self Cite

View full text Add to dashboard Cite

Microinjection is a technique used for transgenesis, mutagenesis, cell labeling, cryopreservation, and in vitro fertilization in multiple single and multicellular organisms. Microinjection requires specialized skills and involves rate limiting and labor-intensive preparatory steps. Here we constructed a machine vision guided generalized robot that fully automates the process of microinjection in fruit fly (Drosophila melanogaster) and zebrafish (Danio rerio) embryos. The robot uses machine learning models trained to detect embryos in images of agar plates and identify specific anatomical locations within each embryo in 3D space using dual view microscopes. The robot then serially performs a microinjection in each detected embryo. We constructed and used three such robots to automatically microinject tens of thousands of Drosophila and zebrafish embryos. We systematically optimized robotic microinjection for each species and performed routine transgenesis with proficiency comparable to highly skilled human practitioners while achieving up to 4x increases in microinjection throughput in Drosophila. The robot was utilized to microinject pools of over 20,000 uniquely barcoded plasmids into 1,713 embryos in two days to rapidly generate more than 400 unique transgenic Drosophila lines. This experiment enabled a novel measurement of the number of independent germline integration events per successfully injected embryo. Finally, we showed that robotic microinjection of cryoprotective agents in zebrafish embryos significantly improves vitrification rates and survival of cryopreserved embryos post-thaw as compared to manual microinjection. We anticipate that the robot can be used to carry out microinjection for genome-wide manipulation and cryopreservation at scale in a wide range of organisms.

show abstract

Ultra Rapid Laser Warming For Preservation of Fish and Other Aquatic Species

Cited by 3 publications

References 0 publications

Conduction‐Dominated Cryomesh for Organism Vitrification

Conduction‐Dominated Cryomesh for Organism Vitrification

A machine vision guided robot for fully automated embryonic microinjection

High-throughput genetic manipulation of multicellular organisms using a machine-vision guided embryonic microinjection robot

Contact Info

Product

Resources

About