Automatic reagent handling and assay processing of human biospecimens inside a transportation container for a medical disaster response against radiation

Biological materials can be shipped off-site for diagnostic, therapeutic and research purposes. They usually are kept in certain environments for their final application during transportation. However, active reagent handling during transportation from a collection site to a laboratory or biorepository has not been reported yet. In this paper, we show the application of a micro-controlled centrifugal microfluidic system inside a shipping container that can add reagent to an actively cultured human blood sample during transportation to ensure a rapid biodosimetry of cytokinesis-block micronucleus (CBMN) assay. The newly demonstrated concept could have a significant impact on rapid biodosimetry triage for medical countermeasure in a radiological disaster. It also opens a new capability in accelerated sample processing during transportation for biomedical and healthcare applications.</jats:p

Automatic reagent handling and assay processing of human biospecimens inside a transportation container for a medical disaster response against radiation.

Biological materials can be shipped off-site for diagnostic, therapeutic and research purposes. They usually are kept in certain environments for their final application during transportation. However, active reagent handling during transportation from a collection site to a laboratory or biorepository has not been reported yet. In this paper, we show the application of a micro-controlled centrifugal microfluidic system inside a shipping container that can add reagent to an actively cultured human blood sample during transportation to ensure a rapid biodosimetry of cytokinesis-block micronucleus (CBMN) assay. The newly demonstrated concept could have a significant impact on rapid biodosimetry triage for medical countermeasure in a radiological disaster. It also opens a new capability in accelerated sample processing during transportation for biomedical and healthcare applications

Schematics of the micropipette parameters for cyt-B centrifugal release.

Schematics of the micropipette parameters for cyt-B centrifugal release.</p

A typical image of a binucleated cell with micronuclei.

(TIF)</p

The Smart Shipping Incubator (SSI).

(A) The custom iQ5 shipping incubator loaded with the centrifugal system; (B) Schematics of the centrifugal system: (C) a photo of the Spin Disk Assembly (SDA); (D) a photo of the Motor Assembly (MA); (E) a photo of a “Smart” Cap inserted into a sample tube loaded to a tube holder; (F) a photo of a glass micropipette loaded with cyt-B reagent.</p

Assessing the ability of the 40 μm micropipette with fluorocoating against premature reagent release under mechanical shocks.

(A) top view of the SDA with mounted 25g and 50g shock indictors at different locations. (B) Maximum g-force of the shock indicator triggered for each condition from three trials. (C) Reagent volumes inside the micropipettes under different conditions demonstrate the resistance to premature reagent release by the micropipette valve.</p

Fig 7 -

(A) CBMN dose curves by SSI and conventional process showing strong dose-dependent MN/BN ratios. (B) Pearson’s correlation analysis showing a strong positive correlation between the SSI and the conventional control samples with a coefficient P = 0.9972 and 95% confidence interval (0.9540, 0.9998).</p

Experimental characterization of cyt-B release at different RCF for micropipettes with different orifices (20 and 40 μm) and with a fluorocoating (40 μm).

Experimental characterization of cyt-B release at different RCF for micropipettes with different orifices (20 and 40 μm) and with a fluorocoating (40 μm).</p

The number of BN cells under three culture conditions: In SSI with and without HEPES buffer, and in a traditional CO₂ incubator.

*** represents a p-value less than 0.001 and **** represents a p-value less than 0.0001. “n.s.” stands for “not significant” (p-value larger than 0.0167).</p