3D-printed microplate inserts for long term high-resolution imaging of live brain organoids

Mariana Oksdath Mansilla; Camilo Salazar-Hernandez; Sally L. Perrin; Kaitlin G. Scheer; Gokhan Cildir; John Toubia; Kristyna Sedivakova; Melinda N. Tea; Sakthi Lenin; Elise Ponthier; Erica C.F. Yeo; Vinay Tergaonkar; Santosh Poonnoose; Rebecca Ormsby; Stuart M. Pitson; Michael P. Brown; Lisa M. Ebert; Guillermo Gomez

doi:10.1186/s42490-021-00049-5

3D-printed microplate inserts for long term high-resolution imaging of live brain organoids

Mariana Oksdath Mansilla, Camilo Salazar-Hernandez, Sally L. Perrin, Kaitlin G. Scheer, Gokhan Cildir, John Toubia, Kristyna Sedivakova, Melinda N. Tea, Sakthi Lenin, Elise Ponthier, Erica C.F. Yeo, Vinay Tergaonkar, Santosh Poonnoose, Rebecca Ormsby, Stuart M. Pitson, Michael P. Brown, Lisa M. Ebert, Guillermo Gomez

College of Medicine and Public Health

Research output: Contribution to journal › Article › peer-review

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Abstract

Background
Organoids are a reliable model used in the study of human brain development and under pathological conditions. However, current methods for brain organoid culture generate tissues that range from 0.5 to 2 mm of size, which need to be constantly agitated to allow proper oxygenation. The culture conditions are, therefore, not suitable for whole-brain organoid live imaging, required to study developmental processes and disease progression within physiologically relevant time frames (i.e. days, weeks, months).

Results
Here we designed 3D-printed microplate inserts adaptable to standard 24 multi-well plates, which allow the growth of multiple organoids in pre-defined and fixed XYZ coordinates. This innovation facilitates high-resolution imaging of whole-cerebral organoids, allowing precise assessment of organoid growth and morphology, as well as cell tracking within the organoids, over long periods. We applied this technology to track neocortex development through neuronal progenitors in brain organoids, as well as the movement of patient-derived glioblastoma stem cells within healthy brain organoids.

Conclusions
This new bioengineering platform constitutes a significant advance that permits long term detailed analysis of whole-brain organoids using multimodal inverted fluorescence microscopy.

Original language	English
Article number	6
Number of pages	14
Journal	BMC Biomedical Engineering
Volume	3
DOIs	https://doi.org/10.1186/s42490-021-00049-5
Publication status	Published - Apr 2021

Keywords

3D-Printed Microplates
High-Resolution Imaging
Live Brain Organoids

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10.1186/s42490-021-00049-5Licence: CC BY

Published versionFinal published version, 6.59 MBLicence: CC BY

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Mansilla, M. O., Salazar-Hernandez, C., Perrin, S. L., Scheer, K. G., Cildir, G., Toubia, J., Sedivakova, K., Tea, M. N., Lenin, S., Ponthier, E., Yeo, E. C. F., Tergaonkar, V., Poonnoose, S., Ormsby, R., Pitson, S. M., Brown, M. P., Ebert, L. M., & Gomez, G. (2021). 3D-printed microplate inserts for long term high-resolution imaging of live brain organoids. BMC Biomedical Engineering, 3, Article 6. https://doi.org/10.1186/s42490-021-00049-5

@article{665e4fa415ac4a7c8da1a546df6da9f5,

title = "3D-printed microplate inserts for long term high-resolution imaging of live brain organoids",

abstract = "BackgroundOrganoids are a reliable model used in the study of human brain development and under pathological conditions. However, current methods for brain organoid culture generate tissues that range from 0.5 to 2 mm of size, which need to be constantly agitated to allow proper oxygenation. The culture conditions are, therefore, not suitable for whole-brain organoid live imaging, required to study developmental processes and disease progression within physiologically relevant time frames (i.e. days, weeks, months).ResultsHere we designed 3D-printed microplate inserts adaptable to standard 24 multi-well plates, which allow the growth of multiple organoids in pre-defined and fixed XYZ coordinates. This innovation facilitates high-resolution imaging of whole-cerebral organoids, allowing precise assessment of organoid growth and morphology, as well as cell tracking within the organoids, over long periods. We applied this technology to track neocortex development through neuronal progenitors in brain organoids, as well as the movement of patient-derived glioblastoma stem cells within healthy brain organoids.ConclusionsThis new bioengineering platform constitutes a significant advance that permits long term detailed analysis of whole-brain organoids using multimodal inverted fluorescence microscopy.",

keywords = "3D-Printed Microplates, High-Resolution Imaging, Live Brain Organoids",

author = "Mansilla, {Mariana Oksdath} and Camilo Salazar-Hernandez and Perrin, {Sally L.} and Scheer, {Kaitlin G.} and Gokhan Cildir and John Toubia and Kristyna Sedivakova and Tea, {Melinda N.} and Sakthi Lenin and Elise Ponthier and Yeo, {Erica C.F.} and Vinay Tergaonkar and Santosh Poonnoose and Rebecca Ormsby and Pitson, {Stuart M.} and Brown, {Michael P.} and Ebert, {Lisa M.} and Guillermo Gomez",

year = "2021",

month = apr,

doi = "10.1186/s42490-021-00049-5",

language = "English",

volume = "3",

journal = "BMC Biomedical Engineering",

issn = "2524-4426",

publisher = "Springer Nature",

}

Mansilla, MO, Salazar-Hernandez, C, Perrin, SL, Scheer, KG, Cildir, G, Toubia, J, Sedivakova, K, Tea, MN, Lenin, S, Ponthier, E, Yeo, ECF, Tergaonkar, V, Poonnoose, S , Ormsby, R, Pitson, SM, Brown, MP, Ebert, LM & Gomez, G 2021, '3D-printed microplate inserts for long term high-resolution imaging of live brain organoids', BMC Biomedical Engineering, vol. 3, 6. https://doi.org/10.1186/s42490-021-00049-5

TY - JOUR

T1 - 3D-printed microplate inserts for long term high-resolution imaging of live brain organoids

AU - Mansilla, Mariana Oksdath

AU - Salazar-Hernandez, Camilo

AU - Perrin, Sally L.

AU - Scheer, Kaitlin G.

AU - Cildir, Gokhan

AU - Toubia, John

AU - Sedivakova, Kristyna

AU - Tea, Melinda N.

AU - Lenin, Sakthi

AU - Ponthier, Elise

AU - Yeo, Erica C.F.

AU - Tergaonkar, Vinay

AU - Poonnoose, Santosh

AU - Ormsby, Rebecca

AU - Pitson, Stuart M.

AU - Brown, Michael P.

AU - Ebert, Lisa M.

AU - Gomez, Guillermo

PY - 2021/4

Y1 - 2021/4

N2 - BackgroundOrganoids are a reliable model used in the study of human brain development and under pathological conditions. However, current methods for brain organoid culture generate tissues that range from 0.5 to 2 mm of size, which need to be constantly agitated to allow proper oxygenation. The culture conditions are, therefore, not suitable for whole-brain organoid live imaging, required to study developmental processes and disease progression within physiologically relevant time frames (i.e. days, weeks, months).ResultsHere we designed 3D-printed microplate inserts adaptable to standard 24 multi-well plates, which allow the growth of multiple organoids in pre-defined and fixed XYZ coordinates. This innovation facilitates high-resolution imaging of whole-cerebral organoids, allowing precise assessment of organoid growth and morphology, as well as cell tracking within the organoids, over long periods. We applied this technology to track neocortex development through neuronal progenitors in brain organoids, as well as the movement of patient-derived glioblastoma stem cells within healthy brain organoids.ConclusionsThis new bioengineering platform constitutes a significant advance that permits long term detailed analysis of whole-brain organoids using multimodal inverted fluorescence microscopy.

AB - BackgroundOrganoids are a reliable model used in the study of human brain development and under pathological conditions. However, current methods for brain organoid culture generate tissues that range from 0.5 to 2 mm of size, which need to be constantly agitated to allow proper oxygenation. The culture conditions are, therefore, not suitable for whole-brain organoid live imaging, required to study developmental processes and disease progression within physiologically relevant time frames (i.e. days, weeks, months).ResultsHere we designed 3D-printed microplate inserts adaptable to standard 24 multi-well plates, which allow the growth of multiple organoids in pre-defined and fixed XYZ coordinates. This innovation facilitates high-resolution imaging of whole-cerebral organoids, allowing precise assessment of organoid growth and morphology, as well as cell tracking within the organoids, over long periods. We applied this technology to track neocortex development through neuronal progenitors in brain organoids, as well as the movement of patient-derived glioblastoma stem cells within healthy brain organoids.ConclusionsThis new bioengineering platform constitutes a significant advance that permits long term detailed analysis of whole-brain organoids using multimodal inverted fluorescence microscopy.

KW - 3D-Printed Microplates

KW - High-Resolution Imaging

KW - Live Brain Organoids

U2 - 10.1186/s42490-021-00049-5

DO - 10.1186/s42490-021-00049-5

M3 - Article

SN - 2524-4426

VL - 3

JO - BMC Biomedical Engineering

JF - BMC Biomedical Engineering

M1 - 6

ER -

3D-printed microplate inserts for long term high-resolution imaging of live brain organoids

Abstract

Keywords

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