Clinical application of mixed reality holographic imaging technology in scaling and root planing of severe periodontitis: a proof of concept.

OBJECTIVES: To demonstrate the potential application of mixed reality (MR) holographic imaging technology in subgingival scaling and root planing (SRP) for patient with advanced periodontitis. METHODS: This case series comprised the analysis of 1566 sites from 261 teeth of 10 patients with advanced periodontitis. Digital CBCT scans and intraoral scans of the patients were digitally acquired preoperatively and aligned to create a three-dimensional periodontal visualization model. Through rendering, interactive holographic images were displayed using MR. The surgeon first used MR images to communicate with the patients, and then facilitated SRP under their guidance. Probing pocket depth (PPD), clinical attachment loss (CAL), Plaque index (PI), and bleeding on probing (BOP) parameters were recorded at baseline and at 8-week postoperatively. Patient-reported outcome indicator questionnaires on self-efficacy were also collected. RESULTS: PPD, CAL, PI, and BOP significantly decreased at 8-week following MR hologram-assisted SRP (p<0.001). For sites with PPD≥4 mm, PPD and CAL declined by 2.33±1.23 mm and 0.69±1.07 mm, respectively. PI significantly decreased from 1.94±0.61 to 0.82±0.58 (p < 0.001) and BOP sites decreased significantly from 84.11% to 40.25%. After receiving MR holograms for condition communication, most patients had a better perception of the effectiveness of SRP treatment and the benefits it brings. 80% of the subjects expressed their willingness to undergo MR-assisted periodontal treatment in the future. CONCLUSION: These results provide preliminary support for MR hologram-assisted digital SRP. With this technology, images of the gingiva and alveolar bone can be displayed in real time, accurately and three-dimensionally. This improves SRP effectiveness, diminishes complications, and enhances patients' confidence in the treatment. CLINICAL SIGNIFICANCE: MR holographic imaging-based digital SRP is a clinically feasible and promising adjunctive periodontal treatment option. It may contribute to improved non-surgical treatment efficacy in patients with severe periodontitis.

Multiscale Materials Engineering via Self-Assembly of Pentapeptide Derivatives from SARS CoV E Protein.

Short peptide-based supramolecular hydrogels hold enormous potential for a wide range of applications. However, the gelation of these systems is very challenging to control. Minor changes in the peptide sequence can significantly influence the self-assembly mechanism and thereby the gelation propensity. The involvement of SARS CoV E protein in the assembly and release of the virus suggests that it may have inherent self-assembling properties that can contribute to the development of hydrogels. Here, three pentapeptide sequences derived from C-terminal of SARS CoV E protein are explored with same amino acid residues but different sequence distributions and discovered a drastic difference in the gelation propensity. By combining spectroscopic and microscopic techniques, the relationship between peptide sequence arrangement and molecular assembly structure are demonstrated, and how these influence the mechanical properties of the hydrogel. The present study expands the variety of secondary structures for generating supramolecular hydrogels by introducing the 310-helix as the primary building block for gelation, facilitated by a water-mediated structural transition into ß-sheet conformation. Moreover, these Fmoc-modified pentapeptide hydrogels/supramolecular assemblies with tunable morphology and mechanical properties are suitable for tissue engineering, injectable delivery, and 3D bio-printing applications.

Application of Digital Holographic Imaging to Monitor Real-Time Cardiomyocyte Hypertrophy Dynamics in Response to Norepinephrine Stimulation.

Cardiomyocyte hypertrophy, characterized by an increase in cell size, is associated with various cardiovascular diseases driven by factors including hypertension, myocardial infarction, and valve dysfunction. In vitro primary cardiomyocyte culture models have yielded numerous insights into the intrinsic and extrinsic mechanisms driving hypertrophic growth. However, due to limitations in current approaches, the dynamics of cardiomyocyte hypertrophic responses remain poorly characterized. In this study, we evaluate the application of the Holomonitor M4 digital holographic imaging microscope to track dynamic changes in cardiomyocyte surface area and volume in response to norepinephrine treatment, a model hypertrophic stimulus. The Holomonitor M4 permits non-invasive, label-free imaging of three-dimensional changes in cell morphology with minimal phototoxicity, thus enabling long-term imaging studies. Untreated and norepinephrine-stimulated primary neonatal rat cardiomyocytes were live-imaged on the Holomonitor M4, which was followed by image segmentation and single-cell tracking using the HOLOMONITOR App Suite software version 4.0.1.546. The 24 h treatment of cultured cardiomyocytes with norepinephrine increased cardiomyocyte spreading and optical volume as expected, validating the reliability of the approach. Single-cell tracking of both cardiomyocyte surface area and three-dimensional optical volume revealed dynamic increases in these parameters throughout the 24 h imaging period, demonstrating the potential of this technology to explore cardiomyocyte hypertrophic responses with greater temporal resolution; however, technological limitations were also observed and should be considered in the experimental design and interpretation of results. Overall, leveraging the unique advantages of the Holomonitor M4 digital holographic imaging system has the potential to empower future work towards understanding the molecular and cellular mechanisms underlying cardiomyocyte hypertrophy with enhanced temporal clarity.

Comparison of holotomographic microscopy and coherence-controlled holographic microscopy.

Quantitative phase imaging (QPI) is a powerful tool for label-free visualisation of living cells. Here, we compare two QPI microscopes - the Telight Q-Phase microscope and the Nanolive 3D Cell Explorer-fluo microscope. Both systems provide unbiased information about cell morphology, such as individual cell dry mass, perimeter and area. The Q-Phase microscope uses artefact-free, coherence-controlled holographic imaging technology to visualise cells in real time with minimal phototoxicity. The 3D Cell Explorer-fluo employs laser-based holotomography to reconstruct 3D images of living cells, visualising their internal structures and dynamics. Here, we analysed the strengths and limitations of both microscopes when examining two morphologically distinct cell lines - the cuboidal epithelial MDCK cells which form multicellular clusters and solitary growing Rat2 fibroblasts. We focus mainly on the ability of the devices to generate images suitable for single-cell segmentation by the built-in software, and we discuss the segmentation results and quantitative data generated from the segmented images. We show that both microscopes offer slightly different advantages, and the choice between them depends on the specific requirements and goals of the user.

Refractive Index Imaging Reveals That Elimination of the ATP Synthase C Subunit Does Not Prevent the Adenine Nucleotide Translocase-Dependent Mitochondrial Permeability Transition.

The mitochondrial permeability transition pore (mPTP) is a large, weakly selective pore that opens in the mitochondrial inner membrane in response to the pathological increase in matrix Ca2+ concentration. mPTP activation has been implicated as a key factor contributing to stress-induced necrotic and apoptotic cell death. The molecular identity of the mPTP is not completely understood. Both ATP synthase and adenine nucleotide translocase (ANT) have been described as important components of the mPTP. Using a refractive index (RI) imaging approach, we recently demonstrated that the removal of either ATP synthase or ANT eliminates the Ca2+-induced mPTP in experiments with intact cells. These results suggest that mPTP formation relies on the interaction between ATP synthase and ANT protein complexes. To gain further insight into this process, we used RI imaging to investigate mPTP properties in cells with a genetically eliminated C subunit of ATP synthase. These cells also lack ATP6, ATP8, 6.8PL subunits and DAPIT but, importantly, have a vestigial ATP synthase complex with assembled F1 and peripheral stalk domains. We found that these cells can still undergo mPTP activation, which can be blocked by the ANT inhibitor bongkrekic acid. These results suggest that ANT can form the pore independently from the C subunit but still requires the presence of other components of ATP synthase.

Cancer metabolic features allow discrimination of tumor from white blood cells by label-free multimodal optical imaging.

Circulating tumor cells (CTCs) are tumor cells that have penetrated the circulatory system preserving tumor properties and heterogeneity. Detection and characterization of CTCs has high potential clinical values and many technologies have been developed for CTC identification. These approaches remain challenged by the extraordinary rarity of CTCs and the difficulty of efficiently distinguishing cancer from the much larger number of white blood cells in the bloodstream. Consequently, there is still a need for efficient and rapid methods to capture the broad spectrum of tumor cells circulating in the blood. Herein, we exploit the peculiarities of cancer metabolism for discriminating cancer from WBCs. Using deuterated glucose and Raman microscopy we show that a) the known ability of cancer cells to take up glucose at greatly increased rates compared to non-cancer cells results in the lipid generation and accumulation into lipid droplets and, b) by contrast, leukocytes do not appear to generate visible LDs. The difference in LD abundance is such that it provides a reliable parameter for distinguishing cancer from blood cells. For LD sensitive detections in a cell at rates suitable for screening purposes, we test a polarization-sensitive digital holographic imaging (PSDHI) technique that detects the birefringent properties of the LDs. By using polarization-sensitive digital holographic imaging, cancer cells (prostate cancer, PC3 and hepatocarcinoma cells, HepG2) can be rapidly discriminated from leukocytes with reliability close to 100%. The combined Raman and PSDHI microscopy platform lays the foundations for the future development of a new label-free, simple and universally applicable cancer cells' isolation method.

Caspase-9-mediated cleavage of vimentin attenuates the aggressiveness of leukemic NB4 cells.

Vimentin is a main type 3 intermediate filament protein. It seems that abnormal expression of vimentin is contributed to the appearance of the aggressive feature of cancer cells. So that it has been reported that malignancy and epithelial-mesenchymal transition in solid tumors, and poor clinical outcomes in patients with lymphocytic leukemia and acute myelocytic leukemia have been associated with the high expression of vimentin. Vimentin is a non-caspase substrate of caspase-9 although its cleavage by caspase-9 in biological processes has not been reported. In the present study, we sought to understand whether vimentin cleavage mediated by caspase-9 could reverse the malignancy in leukemic cells. Herein, to address the issue, we investigated vimentin changes in differentiation and took advantage of the inducible caspase-9 (iC9)/AP1903 system in human leukemic NB4 cells. Following the transfection and treatment of the cells using the iC9/AP1903 system, vimentin expression, cleavage, and subsequently, the cell invasion and the relevant markers such as CD44 and MMP-9 were evaluated. Our results revealed the downregulation and cleavage of vimentin which attenuates the malignant phenotype of the NB4 cells. Considering the favorable effect of this strategy in keeping down the malignant features of the leukemic cells, the effect of the iC9/AP1903 system in combination with all-trans-retinoic acid (ATRA) treatment was evaluated. The obtained data prove that iC9/AP1903 significantly makes the leukemic cells more sensitive to ATRA.

The PERCIVAL detector: first user experiments.

The PERCIVAL detector is a CMOS imager designed for the soft X-ray regime at photon sources. Although still in its final development phase, it has recently seen its first user experiments: ptychography at a free-electron laser, holographic imaging at a storage ring and preliminary tests on X-ray photon correlation spectroscopy. The detector performed remarkably well in terms of spatial resolution achievable in the sample plane, owing to its small pixel size, large active area and very large dynamic range; but also in terms of its frame rate, which is significantly faster than traditional CCDs. In particular, it is the combination of these features which makes PERCIVAL an attractive option for soft X-ray science.

Portable, Automated and Deep-Learning-Enabled Microscopy for Smartphone-Tethered Optical Platform Towards Remote Homecare Diagnostics: A Review.

Globally new pandemic diseases induce urgent demands for portable diagnostic systems to prevent and control infectious diseases. Smartphone-based portable diagnostic devices are significantly efficient tools to user-friendly connect personalized health conditions and collect valuable optical information for rapid diagnosis and biomedical research through at-home screening. Deep learning algorithms for portable microscopes also help to enhance diagnostic accuracy by reducing the imaging resolution gap between benchtop and portable microscopes. This review highlighted recent progress and continued efforts in a smartphone-tethered optical platform through portable, automated, and deep-learning-enabled microscopy for personalized diagnostics and remote monitoring. In detail, the optical platforms through smartphone-based microscopes and lens-free holographic microscopy are introduced, and deep learning-based portable microscopic imaging is explained to improve the image resolution and accuracy of diagnostics. The challenges and prospects of portable optical systems with microfluidic channels and a compact microscope to screen COVID-19 in the current pandemic are also discussed. It has been believed that this review offers a novel guide for rapid diagnosis, biomedical imaging, and digital healthcare with low cost and portability.

Application evaluation of mixed-reality holographic imaging technology in the surgical treatment of spinal cord glioma.

Objectives: To investigate the application effect of mixed reality (MR) holographic imaging technology in the clinical surgical treatment of spinal cord glioma. Methods: The clinical data of 53 patients with spinal cord glioma who underwent surgical treatment in the Neurosurgery Department of our hospital from January 2017 to May 2020 were retrospectively studied. All the patients were divided into two groups according to different assistive technologies during the surgery: the observation group and the control group, with 30 cases and 23 cases respectively. Patients in the observation group received MR holographic imaging technology intraoperatively, while those in the control group received ultrasound. The surgical conditions of the two groups: the rate of complete resection of tumor lesions and the evaluation accuracy of complete resection were compared. Patients were followed up for 12 months in the outpatient department after surgery, and the recovery of postoperative spinal physiological function was evaluated based on imaging review and MMS scale grading, and the recurrence was obtained. Results: There was no statistical significance in the basic clinical conditions between the two groups (P>0.05), and the total tumor resection rate in the experimental group was 96.67%, and that in the control group was 82.61%, showing a statistically significant difference (P<0.05). Based on enhanced MRI examination as the standard, the evaluation accuracy of intraoperative complete tumor resection in the experimental group was 93.33%, significantly higher than that in the control group (73.54%), with a statistical significance (P<0.05). The incidence of postoperative complications was 3.33% in the experimental group and 21.74% in the control group, with a statistically significant difference (P<0.05). Postoperative follow-up showed that good recovery rate of spinal cord function in the experimental group was 56.70%, and that in the control group was 41.09%, with a statistically significant difference (P<0.05). The recurrence rate was 0 in the experimental group and 4.34% in the control group at follow-up, with no statistically significant difference (P>0.05). Conclusions: With the application of MR holographic imaging technology in the surgical treatment of spinal cord glioma, critical clinical value can be realized. Specifically, the resection degree of spinal cord glioma can be displayed in real time, accurately, and three-dimensionally, the effect of surgical resection can be improved, surgical complications can be diminished, and the recovery of spinal cord function can be accelerated.

The Hippo pathway drives the cellular response to hydrostatic pressure.

Cells need to rapidly and precisely react to multiple mechanical and chemical stimuli in order to ensure precise context-dependent responses. This requires dynamic cellular signalling events that ensure homeostasis and plasticity when needed. A less well-understood process is cellular response to elevated interstitial fluid pressure, where the cell senses and responds to changes in extracellular hydrostatic pressure. Here, using quantitative label-free digital holographic imaging, combined with genome editing, biochemical assays and confocal imaging, we analyse the temporal cellular response to hydrostatic pressure. Upon elevated cyclic hydrostatic pressure, the cell responds by rapid, dramatic and reversible changes in cellular volume. We show that YAP and TAZ, the co-transcriptional regulators of the Hippo signalling pathway, control cell volume and that cells without YAP and TAZ have lower plasma membrane tension. We present direct evidence that YAP/TAZ drive the cellular response to hydrostatic pressure, a process that is at least partly mediated via clathrin-dependent endocytosis. Additionally, upon elevated oscillating hydrostatic pressure, YAP/TAZ are activated and induce TEAD-mediated transcription and expression of cellular components involved in dynamic regulation of cell volume and extracellular matrix. This cellular response confers a feedback loop that allows the cell to robustly respond to changes in interstitial fluid pressure.

Combination of holographic imaging with robotic partial nephrectomy for renal hilar tumor treatment.

OBJECTIVES: To evaluate the clinical value of the holographic imaging technology in combination with robotic-assisted partial nephrectomy (RAPN) for renal hilar tumor treatment. PATIENTS AND METHODS: From Dec. 2018 to Dec. 2021, patients diagnosed with renal hilar tumor were included in this retrospective study. Before the surgery, the engineers established the holographic image models based on the enhanced CT data. The models were used in patient consultation, pre-surgery planning and surgery simulation. During the RAPN, the navigation was achieved by real-time overlapping of the holographic images on the robotic surgery endoscopic views. The navigation technique helped the surgeon to identify the important anatomic structures such as tumor, renal vein, renal artery, and pelvis. RESULTS: There were total of eight patients with renal hilar tumor who underwent RAPN combined with holographic imaging technique. The mean age was 57.3 years, the median ASA score was 2. The mean tumor size was 42.4 mm and the median RENAL Nephrometry score was 9.5. The clinical stages were cT1a (37.5%) and cT1b (62.5%). All the procedures were performed uneventfully by one surgeon. The mean operative time was 144.3 min, and the mean warm ischemia time was 27.9 min. The mean estimated blood loss was 86.3 ml. There was no conversion to open surgery or radical nephrectomy. There were no Clavien-Dindo ≥ 3 perioperative complications. CONCLUSIONS: Using the holographic imaging technique, the pre-surgery planning, simulation of renal arterial clamp and excision of the tumor, and intraoperative navigation were feasible and helpful in facilitating RAPN.

Nanosecond timing and synchronization scheme for holographic pump-probe studies at the MID instrument at European XFEL.

Single-pulse holographic imaging at XFEL sources with 1012 photons delivered in pulses shorter than 100 fs reveal new quantitative insights into fast phenomena. Here, a timing and synchronization scheme for stroboscopic imaging and quantitative analysis of fast phenomena on time scales (sub-ns) and length-scales (≲100 nm) inaccessible by visible light is reported. A fully electronic delay-and-trigger system has been implemented at the MID station at the European XFEL, and applied to the study of emerging laser-driven cavitation bubbles in water. Synchronization and timing precision have been characterized to be better than 1 ns.

Ka Band Holographic Imaging System Based on Linear Frequency Modulation Radar.

Millimeter wave (MMW) technology is expanding rapidly into security screening for dangerous items concealed under clothing. It uses safe non-ionizing radiation and penetrates clothing well. We present a new planar system at Ka band for the three-dimensional simultaneous imaging of both sides of an inspected person where the images are produced in real time by a recently proposed generalized holographic reconstruction algorithm. Low-cost linear frequency modulation (LFM) radar technology is used along with a simple but efficient method for system calibration. Experimental characterization of the spatial resolution and the sensitivity of the system prototype has been carried out. It is established that the achieved spatial resolution is 6 mm or better if the item is not obscured by clothing and it may deteriorate to 7 mm depending on the clothing hiding the item. The spatial sensitivity is confirmed to be at least 2 mm.

Application of holographic image navigation in urological laparoscopic and robotic surgery / 中华泌尿外科杂志

Objective To evaluate the clinical value of holographic image navigation in urological laparoscopic and robotic surgery.Methods The data of patients were reviewed retrospectively for whom accepted holographic image navigation laparoscopic and robotic surgery from Jan.2019 to Dec.2019 in Beijing United Family Hospital and other 18 medical centers,including 78 cases of renal tumor,2 cases of bladder cancer,2 cases of adrenal gland tumor,1 cases of renal cyst,1 case of prostate cancer,1 case of sweat gland carcinoma with lymph node metastasis,1 case of pelvic metastasis after radical cystectomy.All the patients underwent operations.In the laparoscopic surgery group,there were 27 cases of partial nephrectomy,1 case of radical prostatectomy,2 cases of radical cystectomy and 2 cases of adrenalectomy.In the da Vinci robotic surgery group of 54 cases,there were 51 cases of partial nephrectomy,1 case of retroperitoneal lymph node dissection,1 case of retroperitoneal bilateral renal cyst deroofing and 1 case of resection of pelvic metastasis.There were 41 partial nephrectomy patients with available clinical data for statistic,with a median age of 53.5 years (range 24-76),including 26 males and 15 females.The median R.E.N.A.L score was 7.8 (range 4-11).Before the operation,the engineers established the holographic image based on the contrast CT images and reports.The surgeon applied the holographic image for preoperative planning.During the operation,the navigation was achieved by real time fusing holographic images with the laparoscopic surgery images in the screen.Results All the procedures had been complete uneventfully.The holographic images helped surgeon in understanding the visual three-dimension structure and relation of vessels supplying tumor or resection tissue,lymph nodes and nerves.By manipulating the holographic images extracorporeally,the fused image guide surgeons about location vessel,lymph node and other important structure and then facilitate the delicate dissection.For the 41 cases with available clinical data including 23 cases of robotic-assisted partial nephrectomy and 18 cases of laparoscopic nephrectomy,the median operation time was 140 (range 50-225) min,the median warm ischemia time was 23 (range 14-60) min,the median blood loss was 80(range 5-1 200) ml.In the robotic surgery group,the median operation time was 140 (range 50-215)min,the median warm i schemia time was 21 (range 17-40)min,the median blood loss was 150(range 30-1 200)ml.In the laparoscopic surgery group,the median operation time was 160(range 80-225)min,the median warm ischemia time was 25 (range 14-60)min,the median blood loss was 50 (range 5-1 200) ml.All the patients had no adjacent organ injury during operation.There were 2 cases with Clavien Ⅱ complications.One required transfusion and the other one suffered hematoma post-operation.However,the tumors were located in the renal hilus for these 2 cases and the R.E.N.A.L scores were both 11.Conclusions Holographic image navigation can help location and recognize important anatomic structures during the surgical procedures..This technique will reduce the tissue injury,decrease the complications and improve the success rate of surgery.

Digital Holographic Imaging as a Method for Quantitative, Live Cell Imaging of Drug Response to Novel Targeted Cancer Therapies.

Digital holographic imaging (DHI) is a noninvasive, live cell imaging technique that enables long-term quantitative visualization of cells in culture. DHI uses phase-shift imaging to monitor and quantify cellular events such as cell division, cell death, cell migration, and drug responses. In recent years, the application of DHI has expanded from its use in the laboratory to the clinical setting, and currently it is being developed for use in theranostics. Here, we describe the use of the DHI platform HoloMonitorM4 to evaluate the effects of novel, targeted cancer therapies on cell viability and proliferation using the HeLa cancer cell line as a model. We present single cell tracking and population-wide analysis of multiple cell morphology parameters.

Holographic Three-Dimensional Imaging of Terra-Cotta Warrior Model Using Fractional Fourier Transform.

Holographic three-dimensional (3D) imaging of Terra-Cotta Warrior model using Fractional Fourier Transform is introduced in this paper. Phase holograms of Terra-Cotta Warrior model are calculated from 60 horizontal viewing-angles by the use of fractional Fourier transform (FRT). Multiple phase holograms are calculated for each angle by adding proper pseudorandom phase to reduce the speckle noise of a reconstructed image. Experimental system based on high-resolution phase-only spatial light modulator (SLM) is built for 3D image reconstruction from the calculated phase holograms. The texture of the Terra-Cotta Warrior model is rough. The calculation of rough texture is optimized in order to show better model details. The effects of computing distance and layer thickness on imaging quality are analyzed finally.

Near-Field Three-Dimensional Planar Millimeter-Wave Holographic Imaging by Using Frequency Scaling Algorithm.

In this paper, a fast three-dimensional (3-D) frequency scaling algorithm (FSA) with large depth of focus is presented for near-field planar millimeter-wave (MMW) holographic imaging. Considering the cross-range range coupling term which is neglected in the conventional range migration algorithm (RMA), we propose an algorithm performing the range cell migration correction for de-chirped signals without interpolation by using a 3-D frequency scaling operation. First, to deal with the cross-range range coupling term, a 3-D frequency scaling operator is derived to eliminate the space variation of range cell migration. Then, a range migration correction factor is performed to compensate for the residual range cell migration. Finally, the imaging results are obtained by matched filtering in the cross-range direction. Compared with the conventional RMA, the proposed algorithm is comparable in accuracy but more efficient by using only chirp multiplications and fast Fourier transforms (FFTs). The algorithm has been tested with satisfying results by both simulation and experiment.

Wavelength-coded volume holographic imaging endoscope for multidepth imaging.

A wavelength-coded volume holographic imaging (WC-VHI) endoscope system capable of simultaneous multifocal imaging is presented. The system images light from two depths separated by 100 µm in a tissue sample by using axial chromatic dispersion of a gradient index probe in combination with two light-emitting diode sources and a multiplexed volume hologram to separate the images. This system is different from previous VHI systems in that it uses planar multiplexed gratings and does not require curved holographic gratings. This results in improved lateral imaging resolution from 228.1 to 322.5 lp/mm. This letter describes the design and fabrication of the WC-VHI endoscope and experimental images of hard and soft resolution targets and biological tissue samples to illustrate the performance properties.

Applications of label-free, quantitative phase holographic imaging cytometry to the development of multi-specific nanoscale pharmaceutical formulations.

A label-free, high content, time-lapse holographic imaging system was applied to studies in pharmaceutical compound development. Multiple fields of cellular images are obtained over typically several day evaluations within standard CO2 incubators. Events are segmented to obtain population data of cellular features, which are displayed in scattergrams and histograms. Cell tracking is accomplished, accompanied by Cartesian plots of cell movement, as well as plots of cell features vs. time in novel 4-D displays of X position, Y position, time, and cell thickness. Our review of the instrument validation data includes 1) tracking of Giant HeLa cells, which may be undergoing neosis, a process of tumor stem cell generation; 2) tracking the effects of cell cycle related toxic agents on cell lines; 3) using MicroRNAs to reverse the polarization state in macrophages to induce tumor cell killing; 4) development of liposomal nanoformulations to overcome Multi-Drug Resistance (MDR) in ovarian cancer cells; and 5) development of dual sensitive micelles to specifically target matrix metalloproteinase 2 (MMP2) over-expressing cell lines. © 2017 International Society for Advancement of Cytometry.