Laser scanning cytometry (LSC) is a slide-based method that is used clinically for Quantitative Imaging Cytometry (QIC). A “Clatch” slide, named after the inventor, which is used in conjunction with the LSC for immunophenotyping patient cell samples, has several drawbacks. The slide requires time consuming and laborious pipette steps, making the slide prone to handling errors. The Clatch slide also uses a significant amount of cell sample, limiting the number of analyses for fine needle aspirate (FNA) samples. This thesis details an automated microfluidic system, composed of an embedded circuit, a plastic and polymer microfluidic device, and an aluminum frame, which can perform the same immunophenotyping procedures. This new system reduces the labor from 36 pipette steps to 8, it reduces the amount of cell sample from 180 μL to 56 μL, and it shortens the entire procedure from 75 minutes to 42 minutes.

The effect of decompression stress on circulating microparticles (MPs) from leukocytes (LMP), platelets (PMP), and endothelial cells (EMP) was investigated in fifteen male naval clearance divers. Venous blood samples were obtained 30 min before and 75 min after exposure to 81msw for 20 min. MPs were isolated by differential centrifugation and immunophenotyped using multiparameter flow cytometry. Venous gas emboli (VGE) were assessed using Doppler ultrasound every 40 min post-dive and subsequently graded using the Kisman Integrated Severity Score (mean KISS=21.92, indicating moderate level of VGE). Following the dive there was increased expression of CD41a, CD106, CD62P and CD31 on MPs, while CD45 and CD141 expression decreased. A positive correlation was found between KISS and CD41a expression post-dive. These results indicate that decompression stress activated platelets, producing PMPs and resulting in potential vascular disruption or injury. The inclusion of MP measures in future DCS-related research may help identify biomarkers of DCS.

Laser scanning cytometry (LSC) is a slide-based method that is used clinically for Quantitative Imaging Cytometry (QIC). A “Clatch” slide, named after the inventor, which is used in conjunction with the LSC for immunophenotyping patient cell samples, has several drawbacks. The slide requires time consuming and laborious pipette steps, making the slide prone to handling errors. The Clatch slide also uses a significant amount of cell sample, limiting the number of analyses for fine needle aspirate (FNA) samples. This thesis details an automated microfluidic system, composed of an embedded circuit, a plastic and polymer microfluidic device, and an aluminum frame, which can perform the same immunophenotyping procedures. This new system reduces the labor from 36 pipette steps to 8, it reduces the amount of cell sample from 180 μL to 56 μL, and it shortens the entire procedure from 75 minutes to 42 minutes.

The effect of decompression stress on circulating microparticles (MPs) from leukocytes (LMP), platelets (PMP), and endothelial cells (EMP) was investigated in fifteen male naval clearance divers. Venous blood samples were obtained 30 min before and 75 min after exposure to 81msw for 20 min. MPs were isolated by differential centrifugation and immunophenotyped using multiparameter flow cytometry. Venous gas emboli (VGE) were assessed using Doppler ultrasound every 40 min post-dive and subsequently graded using the Kisman Integrated Severity Score (mean KISS=21.92, indicating moderate level of VGE). Following the dive there was increased expression of CD41a, CD106, CD62P and CD31 on MPs, while CD45 and CD141 expression decreased. A positive correlation was found between KISS and CD41a expression post-dive. These results indicate that decompression stress activated platelets, producing PMPs and resulting in potential vascular disruption or injury. The inclusion of MP measures in future DCS-related research may help identify biomarkers of DCS.

Thesis (Ph. D.)--University of Hong Kong, 2001. / Includes bibliographical references (leaves 192-206).

Thesis (Ph. D.)--University of Texas at Austin, 2003. / Vita. Includes bibliographical references. Available also from UMI Company.

Thesis (M. Phil.)--University of Hong Kong, 2001. / Includes bibliographical references (leaves 114-127).

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

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