Toward complete laser ablation of melanoma contaminant cells in a co‐culture outgrowth model via image cytometry

Abstract

Background: Contaminant cancer cells in autologous transplant tissue can cause relapse and the rates are unknown. A method capable of removing all contaminant cells with a high probability detected by cytomic analyses would be useful. Neither 100% cell purging nor techniques for measuring the probability of success have been developed. Here, we report a method for removing 100% of the cells under ideal staining conditions and quantify the probability of success. Methods: Laser ablation was combined with previously reported automated microscopy to purge contaminant cells and evaluate 100% ablation in a co-culture model of prestained mouse melanoma cells mixed with mouse NIH-3T3 cells. Melanoma passage efficiency was measured by: (1) micropipetting single cells into microtiter wells and (2) ablating all but one melanoma cell in co-cultures. Results: (74 6 5)% of single melanoma cells pipetted into microtiter plate wells divided at least once. With abla-

Quantification of Lipid Droplets and Associated Proteins in Cellular Models of Obesity via High-Content/High-Throughput Microscopy and Automated Image Analysis

“…The images are analyzed for information by algorithms designed to identify and extract information relevant to cell/disease model 66–69. Advances in automatic acquisition, measurement, comparison and pattern classification tools for cellular images continue69–77. The concept that digital images from both conventional and confocal microscopy can be analyzed by an evolving set of sophisticated image-analysis algorithms is advancing, not only in the context of chemical and siRNA screening, but, also in the context of modern quantitative approaches to microscopy-based cellular phenotypic characterization78, 79.…”

Slide‐based cytometry for cytomics—A minireview

“…In SBC, by contrast, mechanical or optical cell isolation techniques are of relatively low speed. In two articles of this issue, optical cell enrichment and manipulation techniques suitable for high‐throughput are introduced (15, 16).…”

“…Cytomics is bioinformatic knowledge extraction from a large amount of structural and functional information by molecular cell phenotype analysis of tissues, organs, and organisms at the single‐cell level by image or flow cytometry (5). Areas of research and diagnosis with the demand to virtually measure “anything” in the cell include immunophenotyping (7–9), rare cell detection (10) and characterization in the case of stem cells (11–14) and residual tumor cells (15–17), tissue analysis (18–21), and drug discovery (22, 23). Systemic analysis is also a prerequisite for predictive medicine by genomics, proteomics, and cytomics.…”

Cytomics – importance of multimodal analysis of cell function and proliferation in oncology

“…2004; Rippon & Bishop 2004) and residual tumour cells (Szaniszlo et al . 2004, 2006; Shen & Price 2006), histopathology and drug discovery (Van Osta et al . 2006; Van Osta 2006).…”

“…2006). A method capable of removing all contaminant cells, detected by cytomic analyses with a high probability, is combined laser ablation with automated microscopy (Shen & Price 2006). Thus, complete ablation by automated high‐content image cytometry may become feasible in combination with pre‐therapeutic cytomic analysis of unique cellular expression and/or morphological characteristic patterns (Valet & Hoeffkes 2004; Tarnok et al .…”