Figure 2 from PSA Secretion from Single Circulating Tumor Cells of Metastatic Castration-Naïve Prostate Cancer Patients

<p>Workflow schematically illustrating the DLA processing to obtain viable CTCs. (1) DLA enrichment using the reduced enrichment reagent (RER) protocol; (2) staining using anti–PSMA-PE, anti–CD45-APC, and viability dye calcein AM; (3) sorting of calcein<sup>+</sup> and CD45<sup>–</sup> CTCs enriched from the DLA; (4) seeding into the VyCAP nanowells; (5) imaging using a fluorescence inverted scanning microscope to visualize cells captured in the nanowells; (6) PSA secretion from single cells and PSA capture overnight (24 hours) using an anti–PSA-coated membrane and addition of anti–IgG-PE to visualize the imprint; (7) staining of the membrane for visualization of PSA secretion spots; and (8) correlation of PSA spots with cells with the help of imprint and the SPOT software to identify PSA-secreting and non-secreting PSMA<sup>+</sup>/PSMA<sup>−</sup> CTCs.</p>

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Publication Details

Published
2025-08-18
DOI
https://doi.org/10.1158/2767-9764.29929885
Primary Topic
Prostate Cancer Treatment and Research
Type
article
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article

Figure 2 from PSA Secretion from Single Circulating Tumor Cells of Metastatic Castration-Naïve Prostate Cancer Patients

Leon W.M.M. Terstappen, Danny Houtsma, Brigitte C.M. Haberkorn, Michiel Stevens et al.
Prostate Cancer Treatment and Research
article

Figure 2 from PSA Secretion from Single Circulating Tumor Cells of Metastatic Castration-Naïve Prostate Cancer Patients

Leon W.M.M. Terstappen, Danny Houtsma, Brigitte C.M. Haberkorn, Michiel Stevens, Wendy M. van der Deure, Nick Beije, Paul Hamberg, Khrystany T. Isebia, Richell Booijink, Eshwari Dathathri, Peter A.W. te Boekhorst, John W.M. Martens, Martijn P. Lolkema, Fikri Abali, Jaco Kraan, Ruchi Bansal, Joan van den Bosch, Tanja C. van Dijk
article en

Abstract

<p>Workflow schematically illustrating the DLA processing to obtain viable CTCs. (1) DLA enrichment using the reduced enrichment reagent (RER) protocol; (2) staining using anti–PSMA-PE, anti–CD45-APC, and viability dye calcein AM; (3) sorting of calcein<sup>+</sup> and CD45<sup>–</sup> CTCs enriched from the DLA; (4) seeding into the VyCAP nanowells; (5) imaging using a fluorescence inverted scanning microscope to visualize cells captured in the nanowells; (6) PSA secretion from single cells and PSA capture overnight (24 hours) using an anti–PSA-coated membrane and addition of anti–IgG-PE to visualize the imprint; (7) staining of the membrane for visualization of PSA secretion spots; and (8) correlation of PSA spots with cells with the help of imprint and the SPOT software to identify PSA-secreting and non-secreting PSMA<sup>+</sup>/PSMA<sup>−</sup> CTCs.</p>

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Prostate Cancer Treatment and Research
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Figure 2 from PSA Secretion from Single Circulating Tumor Cells of Metastatic Castration-Naïve Prostate Cancer Patients — Leon W.M.M. Terstappen, Danny Houtsma, et al. · (2025) | TGRS Research Map | TGRS