Design and testing of a centrifugal fluidic device for populating microarrays of spheroid cancer cell cultures
Abstract Background In current cancer spheroid culturing methods, the transfer and histological processing of specimens grown in 96-well plates is a time consuming process. A centrifugal fluidic device was developed and tested for rapid extraction of spheroids from a 96-well plate and subsequent dep...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2020-03-01
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| Series: | Journal of Biological Engineering |
| Online Access: | http://link.springer.com/article/10.1186/s13036-020-0228-6 |
| _version_ | 1828467526005161984 |
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| author | Warren Weisler Samuel Miller Shaphan Jernigan Gregory Buckner Matthew Bryant |
| author_facet | Warren Weisler Samuel Miller Shaphan Jernigan Gregory Buckner Matthew Bryant |
| author_sort | Warren Weisler |
| collection | DOAJ |
| description | Abstract Background In current cancer spheroid culturing methods, the transfer and histological processing of specimens grown in 96-well plates is a time consuming process. A centrifugal fluidic device was developed and tested for rapid extraction of spheroids from a 96-well plate and subsequent deposition into a molded agar receiver block. The deposited spheroids must be compact enough to fit into a standard histology cassette while also maintaining a highly planar arrangement. This size and planarity enable histological processing and sectioning of spheroids in a single section. The device attaches directly to a 96-well plate and uses a standard centrifuge to facilitate spheroid transfer. The agar block is then separated from the device and processed. Results Testing of the device was conducted using six full 96-well plates of fixed Pa14C pancreatic cancer spheroids. On average, 80% of spheroids were successfully transferred into the agar receiver block. Additionally, the planarity of the deposited spheroids was evaluated using confocal laser scanning microscopy. This revealed that, on average, the optimal section plane bisected individual spheroids within 27% of their mean radius. This shows that spheroids are largely deposited in a planar fashion. For rare cases where spheroids had a normalized distance to the plane greater than 1, the section plane either misses or captures a small cross section of the spheroid volume. Conclusions These results indicate that the proposed device is capable of a high capture success rate and high sample planarity, thus demonstrating the capabilities of the device to facilitate rapid histological evaluation of spheroids grown in standard 96-well plates. Planarity figures are likely to be improved by adjusting agar block handling prior to imaging to minimize deformation and better preserve the planarity of deposited spheroids. Additionally, investigation into media additives to reduce spheroid adhesion to 96-well plates would greatly increase the capture success rate of this device. |
| first_indexed | 2024-12-11T04:07:31Z |
| format | Article |
| id | doaj.art-70e2cefc692843d7ad224553f0e411a5 |
| institution | Directory Open Access Journal |
| issn | 1754-1611 |
| language | English |
| last_indexed | 2024-12-11T04:07:31Z |
| publishDate | 2020-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Biological Engineering |
| spelling | doaj.art-70e2cefc692843d7ad224553f0e411a52022-12-22T01:21:29ZengBMCJournal of Biological Engineering1754-16112020-03-0114111010.1186/s13036-020-0228-6Design and testing of a centrifugal fluidic device for populating microarrays of spheroid cancer cell culturesWarren Weisler0Samuel Miller1Shaphan Jernigan2Gregory Buckner3Matthew Bryant4Department of Mechanical and Aerospace Engineering, North Carolina State UniversityDepartment of Mechanical and Aerospace Engineering, North Carolina State UniversityDepartment of Mechanical and Aerospace Engineering, North Carolina State UniversityDepartment of Mechanical and Aerospace Engineering, North Carolina State UniversityDepartment of Mechanical and Aerospace Engineering, North Carolina State UniversityAbstract Background In current cancer spheroid culturing methods, the transfer and histological processing of specimens grown in 96-well plates is a time consuming process. A centrifugal fluidic device was developed and tested for rapid extraction of spheroids from a 96-well plate and subsequent deposition into a molded agar receiver block. The deposited spheroids must be compact enough to fit into a standard histology cassette while also maintaining a highly planar arrangement. This size and planarity enable histological processing and sectioning of spheroids in a single section. The device attaches directly to a 96-well plate and uses a standard centrifuge to facilitate spheroid transfer. The agar block is then separated from the device and processed. Results Testing of the device was conducted using six full 96-well plates of fixed Pa14C pancreatic cancer spheroids. On average, 80% of spheroids were successfully transferred into the agar receiver block. Additionally, the planarity of the deposited spheroids was evaluated using confocal laser scanning microscopy. This revealed that, on average, the optimal section plane bisected individual spheroids within 27% of their mean radius. This shows that spheroids are largely deposited in a planar fashion. For rare cases where spheroids had a normalized distance to the plane greater than 1, the section plane either misses or captures a small cross section of the spheroid volume. Conclusions These results indicate that the proposed device is capable of a high capture success rate and high sample planarity, thus demonstrating the capabilities of the device to facilitate rapid histological evaluation of spheroids grown in standard 96-well plates. Planarity figures are likely to be improved by adjusting agar block handling prior to imaging to minimize deformation and better preserve the planarity of deposited spheroids. Additionally, investigation into media additives to reduce spheroid adhesion to 96-well plates would greatly increase the capture success rate of this device.http://link.springer.com/article/10.1186/s13036-020-0228-6 |
| spellingShingle | Warren Weisler Samuel Miller Shaphan Jernigan Gregory Buckner Matthew Bryant Design and testing of a centrifugal fluidic device for populating microarrays of spheroid cancer cell cultures Journal of Biological Engineering |
| title | Design and testing of a centrifugal fluidic device for populating microarrays of spheroid cancer cell cultures |
| title_full | Design and testing of a centrifugal fluidic device for populating microarrays of spheroid cancer cell cultures |
| title_fullStr | Design and testing of a centrifugal fluidic device for populating microarrays of spheroid cancer cell cultures |
| title_full_unstemmed | Design and testing of a centrifugal fluidic device for populating microarrays of spheroid cancer cell cultures |
| title_short | Design and testing of a centrifugal fluidic device for populating microarrays of spheroid cancer cell cultures |
| title_sort | design and testing of a centrifugal fluidic device for populating microarrays of spheroid cancer cell cultures |
| url | http://link.springer.com/article/10.1186/s13036-020-0228-6 |
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