The joint program of excellence between Northeastern University and PHI began in late 2014. Since then HoloMonitor technology has rapidly established itself as a technology of choice for label-free kinetic cellular analysis. The program covers several activities including:
In May 2017, a summary of the scientific achievements was published in a special issue of Cytometry part A, dedicated to holographic microscopy and quantitative phase imaging.
Read the summary here and view the associated CYTO 2017 presentation here.
Poster session at AACR Annual Meeting 2017. View poster presented by Ed Luther, Supervisor Core Imaging and Cytometry Facility at Northeastern University here.
The expansion of our existing collaboration with PHI is the result of productive utilization of HoloMonitor technology for non-invasive real-time cell cycle analysis in living cells. We are now developing state-of-the art applications multiplexing cell cycle and cell motility.
We rely on in vitro cellular cultures to evaluate the effects of the components of multifunctional nano-based formulations under development. We employ an incubator-adapted, label-free holographic imaging cytometer HoloMonitor M4® (Phase Holographic Imaging, Lund, Sweden) to obtain multi-day time-lapse sequences at 5- minute intervals. An automated stage allows hand-free acquisition of multiple fields of view.
A review of HoloMonitor applications: tracking of Giant HeLa cells, which may be undergoing neosis tracking the effects of cell cycle related toxic agents on cell lines; using MicroRNAs to reverse the polarization state in macrophages to induce tumor cell killing development of liposomal nanoformulations to overcome multi-drug resistance in ovarian cancer cells and development of dual sensitive micelles to specifically target matrix metalloproteinase 2
The authors created a nanoparticle formulation against chemotherapy resistant ovarian cancer, loaded with the commonly used cancer drug paclitaxel together with a drug that breaks down the resistance mechanism of cancer cells. Using HoloMonitor, the scientists were able to show that paclitaxel-resistant ovarian cancer cells stopped multiplying when treated with the nanoparticles.
Methods for producing 4D plot were developed. Images from the time course were exported to ImageJ and treated as an image stack, creating a volume rendering of the cellular motion over time of analysis. The brightness threshold was adjusted to view all cells within the imaging area or only the thicker cells, a feature that allows identification of mitosis, apoptosis, and transition between mesenchymal and amoeboid cellular morphologies
Using holomonitor M4, it was found that FA-(C6+Dox)-LP treated hela cells underwent rapid progression to apoptosis (cell suicide) after 21 hours, as evidenced by a drastic drop in cell area after loss of cell membrane integrity.
Holographic assessment using Holomonitor M4 of the co-cultured cells in real-time showed differences in motility and morphology of macrophages with mir-155 ME treated cells showing greater cellular interaction between the two phenotypes.
Our objective is to develop multi-functional nanotechnology-based antitumor drug delivery systems for improving efficacy of treatments and reducing undesirable side effects. The essential part of this process is the development of un-biased quantitative analytical techniques. We are reporting a successful validation of a very high content, medium throughput system in multi-well plates, employing the newly developed holographic imaging cytometer HoloMonitor® M4 for label-free time-lapse cellular analysis.
Macrophage Response to Polarization
Department of Pharmaceutical Sciences
School of Pharmacy
Bouvé College of Health Sciences
140 The Fenway, Room 228/229
60 Huntington Avenue
Boston, Massachusetts 02115