Label-free enumeration, collection and downstream cytological and cytogenetic analysis of circulating tumor cells

Manjima Dhar, Edward Pao, Corinne Renier, Derek E. Go, James Che, Rosita Montoya, Rachel Conrad, Melissa Matsumoto, Kyra Heirich, Melanie Triboulet, Jianyu Rao, Stefanie S. Jeffrey, Edward B. Garon, Jonathan Goldman, Nagesh P. Rao, Rajan Kulkarni, Elodie Sollier-Christen, Dino Di Carlo

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

Circulating tumor cells (CTCs) have a great potential as indicators of metastatic disease that may help physicians improve cancer prognostication, treatment and patient outcomes. Heterogeneous marker expression as well as the complexity of current antibody-based isolation and analysis systems highlights the need for alternative methods. In this work, we use a microfluidic Vortex device that can selectively isolate potential tumor cells from blood independent of cell surface expression. This system was adapted to interface with three protein-marker-free analysis techniques: (i) an in-flow automated image processing system to enumerate cells released, (ii) cytological analysis using Papanicolaou (Pap) staining and (iii) fluorescence in situ hybridization (FISH) targeting the ALK rearrangement. In-flow counting enables a rapid assessment of the cancer-associated large circulating cells in a sample within minutes to determine whether standard downstream assays such as cytological and cytogenetic analyses that are more time consuming and costly are warranted. Using our platform integrated with these workflows, we analyzed 32 non-small cell lung cancer (NSCLC) and 22 breast cancer patient samples, yielding 60 to 100% of the cancer patients with a cell count over the healthy threshold, depending on the detection method used: respectively 77.8% for automated, 60-100% for cytology, and 80% for immunostaining based enumeration.

Original languageEnglish (US)
Article number35474
JournalScientific Reports
Volume6
DOIs
StatePublished - Oct 14 2016
Externally publishedYes

ASJC Scopus subject areas

  • General

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