Automated rare single cell picking with the ALS cellcelector™

Abstract

Molecular analysis of rare single cells like circulating tumor cells (CTCs) from whole blood patient samples bears multiple challenges. One of those challenges is the efficient and ideally loss-free isolation of CTCs over contaminating white and red blood cells. While there is a multitude of commercial and non-commercial systems available for the enrichment of CTCs their cell output does not deliver the purity most molecular analysis methods require. Here we describe the ALS CellCelector™ which can solve this challenge allowing the retrieval of 100% pure single CTCs from blood processed by different upstream enrichment techniques. It is a multifunctional, extremely flexible system for automated screening of cell culture plates, Petri dishes, and microscope slides. Fixed or live single cells or multicellular clusters detected during screening can be picked out of those plates automatically. The complete scan and picking process is fully documented hence allowing highest standardization and reproducibility of all processes. Use of CellCelector allowed the isolation of pure single tumor cells or clusters from liquid biopsies of breast, prostate, ovarian, colorectal, lung, and brain cancers for their subsequent molecular analysis. © 2018 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

CTC Technologies and Tools

“…Addition of fluorescently labeled antibodies recognizing treatment targets to the staining cocktail is relatively easy, although, one is limited by the number of fluorochromes that can be determined simultaneously. Tools to isolate the single cells from CTC enriched cell suspensions are available (39,41,42,56,57). Probing for gene aberrations of CTC by fluorescence in situ hybridization has also been demonstrated and again reiterated their extensive intra-and inter-patient heterogeneity (52)(53)(54)(55).…”

“…For extensive interrogation of the CTC the individual cells will have to be isolated and for molecular characterization the nucleic acids will have to be amplified. Tools to isolate the single cells from CTC enriched cell suspensions are available (39,41,42,56,57). The steps needed to get from blood to single CTC surely will be associated with cell losses and the different steps will have to be optimized to minimize these losses.…”

An Automated Fluorescence-Based Method to Isolate Bone Marrow-Derived Plasma Cells from Rhesus Macaques Using SIVmac239 SOSIP.664

“…Automated micromanipulators systems, like the ALS CellCelector, have been used primarily to isolate circulating tumor cells. 24 , 25 , 26 , 27 However, Ogunniyi et al. 28 employed this methodology in combination with V(D)J gene amplification to evaluate human humoral responses present in blood and mucosal tissue from HIV-infected patients.…”

Self-Seeding Microwells to Isolate and Assess the Viability of Single Circulating Tumor Cells

“…Other systems are available to assess single cells, for example a self-sufficient micro-droplet generation system that facilitates encapsulation, chemical stimulation, and microscopic analysis of viable cells inside droplets [35]. However, many cells are used and it has not been shown that such systems work for rare cell populations like CTC single cell isolation technologies suitable for CTC analysis, including viable cells do exist like DEPArray™ [36], ALS cellselector™ [37] or FACS sorting [38] but no successful CTC cultures have been published yet. A workflow to successfully obtain viable CTC using self-sorting microwells is presented here.…”