Applications

Other interesting applications

We've captured a few other utilities of IncuCyte® from our users to illustrate the breadth, versatility and value of kinetic live cell imaging. Hopefully, this will help inspire new thought and creativity.


Viral Titers and Antiviral Screening

IncuCyte ZOOM® enables fast, efficient and accurate measurement of infectious viral titres. A highly flexible fixed cell analysis protocol suitable for antiviral screening.

A novel method for the measurement of hepatitis C virus infectious titres using the IncuCyte ZOOM and its application to antiviral screening.

Stewart H, Bartlett C, Ross-Thriepland D, Shaw J, Griffin S, Harris M. J Virol Methods. 2015 Jun 15;218:59-65


Viral Replication

IncuCyte™ was used to quantify replication of the Foot-and-mouth disease virus replicon system within mammalian cells. A rapid and facile alternative to RT-qPCR and CAT assays.

FMDV replicons encoding green fluorescent protein are replication competent

Tulloch F, Pathania U, Luke GA, Nicholson J, Stonehouse NJ, Rowlands DJ, Jackson T, Tuthill T, Haas J, Lamond AI, Ryan MD. Journal of Virological Methods, 209:35-40, 2014

Inhibition of the FMDV sub-genomic replicon by RNA aptamers Forrest,

Forrest , Lear , Herod MR, Ryan M, Rowlands DJ, Stonehouse NJ. The Journal of General Virology, 2014 Dec;95(Pt 12):2649-57


Immune Cell Killing

Cancer cell specific antigens are promising targets for tumour immunotherapy. A novel bi-specific T cell engaging protein was used to direct T cells to kill antigen expressing tumour cells. IncuCyte was used to selectively quantify the kinetics of melanoma cell apoptosis.

Bi-specific TCR-anti CD3 redirected T-cell targeting of NY-ESO-1- and LAGE-1-positive tumors.
McCormack E, Adams KJ, Hassan NJ, Kotian A, Lissin NM, Sami M, et al. Cancer Immunol Immunother, 62(4):773–85, 2013


Cell Tracking

IncuCyte time-lapse imaging has been used to track the movements of individual cells within populations. Analysis of IncuCyte images with ImageJ enables measurement of total and vectoral distance covered in addition to cell speed.

N-cadherin expression level modulates integrin-mediated polarity and strongly impacts on the speed and directionality of glial cell migration.
Camand, E., Peglion, F., Osmani, N., Sanson, M. & Etienne-Manneville, S. J. Cell Sci. 125, 844–57 (2012).

Defining Akt actions in muscle differentiation.
Gardner, S., Anguiano, M. & Rotwein, P. Am. J. Physiol. Cell Physiol. 303, C1292–300 (2012).


Cell Clustering Dynamics

IncuCyte was used to quantify the kinetics of T-cell clustering based on size and shape metrics. A hallmark of T-cell activation is the clustering of T-cells via interaction of the LFA-1 integrin with ICAM-1.

ICAM-1-dependent homotypic aggregates regulate CD8 T cell effector function and differentiation during T cell activation.
Zumwalde, N. a, Domae, E., Mescher, M. F. & Shimizu, Y. J. Immunol. 191, 3681–93 (2013).


Cell Fate

IncuCyte reveals marked variability in myoblast proliferation and fate.

Live cell imaging reveals marked variability in myoblast proliferation and fate.
Gross, S. M. & Rotwein, P. Skelet. Muscle 3, 10 (2013).


Spheroid Invasion

IncuCyte High Definition phase contrast imaging enables dynamic visualisation and analysis (ImageJ) of spheroid invasion through 3D biomatrices.

A functional genetic screen reveals new regulators of β1-integrin activity.
Pellinen, T. et al. A. J. Cell Sci. 125, 649–61 (2012).

Chemical biology drug sensitivity screen identifies sunitinib as synergistic agent with disulfiram in prostate cancer cells.
Ketola, K., Kallioniemi, O. & Iljin, K. PLoS One 7, e51470 (2012).

Lysophosphatidic acid and sphingosine-1-phosphate promote morphogenesis and block invasion of prostate cancer cells in three-dimensional organotypic models.
Härmä, V. et al. Oncogene 31, 2075–89 (2012).


Senescence

Kinetic cell senescence measurements were made using IncuCyte to quantify nuclear area and cell number in long term cell cultures.

A high-content cellular senescence screen identifies candidate tumor suppressors, including EPHA3.
Lahtela, J. et al. Cell Cycle 12, 625–34 (2013).

Alternative measurements of cell senescence have been made using the confluence metric to assess proliferation and monitor drug induced senescence & washout.

A non-catalytic role of choline kinase alpha is important in promoting cancer cell survival.
Falcon, S. C. et al.Oncogenesis 2, e38 (2013).


Intracellular Vesicle Dynamics

Use of IncuCyte High Definition phase contrast images to monitor the dynamics of intracellular vesicle formation.

Label-free detection and dynamic monitoring of drug-induced intracellular vesicle formation enabled using a 2-dimensional matched filter.
Aftab, O. et al. Autophagy 10, 57–69 (2014).


Hypoxia

Fluorescence-based HIF-1 analysis. IncuCyte was used to detect accumulation of GFP in a reporter cell line that expresses EGFP under the control of an artificial hypoxia-responsive promoter (HRP).

Induction of mitochondrial dysfunction as a strategy for targeting tumour cells in metabolically compromised microenvironments.
Zhang, X. et al. Nat. Commun. 5, (2014).

Use of IncuCyte to monitor dynamic changes in hypoxia in 3D spheroids using a hypoxia reporter cell line

Loss of cancer drug activity in colon cancer HCT-116 cells during spheroid formation in a new 3-D spheroid cell culture system.
Karlsson, H., Fryknäs, M., Larsson, R. & Nygren, P. Exp. Cell Res. 318, 1577–85 (2012).


Reactive Oxygen Species (ROS)

Intracellular ROS production monitored in real-time using DCFH-DA as an indicator for oxidative stress.

Identification of anti-allergic effect of Clonorchis sinensis-derived protein venom allergen-like proteins (CsVAL).
Jeong, Y.-I. et al. Biochem. Biophys. Res. Commun. (2014). doi:10.1016/j.bbrc.2014.01.189


Colony Formation

Use of IncuCyte to verify the single cell origin of colonies.

Clonogenicity: holoclones and meroclones contain stem cells.
Beaver, C. M., Ahmed, A. & Masters, J. R. PLoS One 9, e89834 (2014).


Anchorage-Independent Growth

IncuCyte used to image 3D anchorage-independent cell growth in polyHEMA-coated wells.

High-throughput 3D screening reveals differences in drug sensitivities between culture models of JIMT1 breast cancer cells.
Hongisto, V. et al. PLoS One 8, e77232 (2013).


Chromosomal Dynamics

IncuCyte was used to visualize chromosomal dynamics and measure mitotic timing in H2BGFP transfected HeLa cells. IncuCyte images were used to measure the time from nuclear envelope break down (NEB) to anaphase.

ATM-mediated Mad1 Serine 214 phosphorylation regulates Mad1 dimerization and the spindle assembly checkpoint.
Yang, C. et al. Carcinogenesis (2014). doi:10.1093/carcin/bgu087


Megakaryocyte Pro-platelet Production

IncuCyte was used to automatically monitor megakaryocyte adherence and proplatelet formation in the presence of different well coatings.

Serum response factor is an essential transcription factor in megakaryocytic maturation.
Halene, S. et al. Blood 116, 1942–50 (2010).


Intracellular F-actin Dynamics

IncuCyte was used to monitor intracellular F-actin dynamics in mouse bone marrow-derived osteoclast cultures.

The phosphoinositide-3-kinase isoform PI3Kβ regulates osteoclast-mediated bone resorption.
Győri, D. et al. Arthritis Rheumatol. 1443, (2014).

No file selected
0%
Stay up-to-date with what's happening at Essen BioScience

© 2017 Essen BioScience

Privacy Policy