Nanoparticle Induced Cell Magneto-Rotation for the Multiplexed Monitoring of Morphology, Stress and Drug Sensitivity of Suspended Single Cancer Cells.

The metastatic process of a cancer relies on the transformation of some of the primary tumor cells into cells capable of migrating through the Extra-Cellular Matrix (ECM), surrounding the tumor, into the bloodstream and the lymph nodes, and then settle in distant tissue, growing new secondary tumors. By identifying, characterizing and quantifying these cells, the progression of cancer in a patient during therapy can be more accurately assessed. Here we describe the development of a new method for quantitative real time monitoring of cell size and morphology, on single live suspended cancer cells, unconfined in three dimensions. The enabling cell magnetorotation (CM) method is made possible by nanoparticle induced cell magnetization. Using a rotating magnetic field, the magnetically labeled cells are actively rotated, then imaged, using a high definition CCD camera. Under proper conditions, the rotation period of a magnetic object is proportional to its shape factor. We demonstrate first that the rotational period, when measured in real-time, can serve to track cellular response to drugs, cytotoxic agents and other chemical stimuli. In addition, while cells are rotated, they exhibit very specific morphological activities, even without a chemical stimulus. Described also is how to multiplex the CM method, to image several dozens to several thousands of cells simultaneously, and using morphology to classify cells into different phenotypic categories, with each phenotype being correlated with malignancy level. The intrinsic tumor heterogeneity, at the cellular level, can be visualized with relationship graphs. Shown is the ability to monitor cell morphological changes over long periods of time, in real time, in order to detect the metastatic potential for heterogeneous populations of cancer cells, using tools from statistical analysis methods. The method relies on unsupervised Machine Learning algorithms which do not require human inputs. Overall it is demonstrated that the CM method can be used as a diagnostic tool to evaluate the phenotypical heterogeneity in a cell population in general, and in a cancer cell population in particular. This fast and high throughput method promises to efficiently assess the efficacy of personalized therapeutic strategies.PHDApplied PhysicsUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/111434/1/relbez_1.pd

Cell-morphodynamic phenotype classification with application to cancer metastasis using cell magnetorotation and machine-learning.

We define cell morphodynamics as the cell's time dependent morphology. It could be called the cell's shape shifting ability. To measure it we use a biomarker free, dynamic histology method, which is based on multiplexed Cell Magneto-Rotation and Machine Learning. We note that standard studies looking at cells immobilized on microscope slides cannot reveal their shape shifting, no more than pinned butterfly collections can reveal their flight patterns. Using cell magnetorotation, with the aid of cell embedded magnetic nanoparticles, our method allows each cell to move freely in 3 dimensions, with a rapid following of cell deformations in all 3-dimensions, so as to identify and classify a cell by its dynamic morphology. Using object recognition and machine learning algorithms, we continuously measure the real-time shape dynamics of each cell, where from we successfully resolve the inherent broad heterogeneity of the morphological phenotypes found in a given cancer cell population. In three illustrative experiments we have achieved clustering, differentiation, and identification of cells from (A) two distinct cell lines, (B) cells having gone through the epithelial-to-mesenchymal transition, and (C) cells differing only by their motility. This microfluidic method may enable a fast screening and identification of invasive cells, e.g., metastatic cancer cells, even in the absence of biomarkers, thus providing a rapid diagnostics and assessment protocol for effective personalized cancer therapy

Nanoparticle Induced Cell Magneto-Rotation: Monitoring Morphology, Stress and Drug Sensitivity of a Suspended Single Cancer Cell

Single cell analysis has allowed critical discoveries in drug testing, immunobiology and stem cell research. In addition, a change from two to three dimensional growth conditions radically affects cell behavior. This already resulted in new observations on gene expression and communication networks and in better predictions of cell responses to their environment. However, it is still difficult to study the size and shape of single cells that are freely suspended, where morphological changes are highly significant. Described here is a new method for quantitative real time monitoring of cell size and morphology, on single live suspended cancer cells, unconfined in three dimensions. The precision is comparable to that of the best optical microscopes, but, in contrast, there is no need for confining the cell to the imaging plane. The here first introduced cell magnetorotation (CM) method is made possible by nanoparticle induced cell magnetization. By using a rotating magnetic field, the magnetically labeled cell is actively rotated, and the rotational period is measured in real-time. A change in morphology induces a change in the rotational period of the suspended cell (e.g. when the cell gets bigger it rotates slower). The ability to monitor, in real time, cell swelling or death, at the single cell level, is demonstrated. This method could thus be used for multiplexed real time single cell morphology analysis, with implications for drug testing, drug discovery, genomics and three-dimensional culturing

Self‐Assembled Magnetic Bead Biosensor for Measuring Bacterial Growth and Antimicrobial Susceptibility Testing

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/93536/1/2477_ftp.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/93536/2/smll_201200110_sm_suppl.pd