Analysis of cell cycle (S phase) without the need of fixation and antibodies

By -

Dr. I. Okkelman, Dr. R. Dmitriev, Prof. D. Papkovsky and T. Foley (Dept. of Anatomy and Neuroscience) have published article describing new method for analysis of cell cycle in live cells. It is based on the discovery that fluorescence lifetime of conventional DNA-staining dye Hoechst 33342 changes upon incorporation of 5-bromo-2-deoxyuridine (BrdU). This allows simple, direct and real-time analysis of live cells, growing in broad range of tissue cultures, from conventional adherent to complex 3D organoid models. The method must be also compatible with in vivo imaging, two-photon excitation and other state-of-the-art imaging platforms and must represent important framework for applications of live cell FLIM in life science areas. It should also inspire other research groups on design of better and more compatible with live cell imaging DNA-binding fluorescent dyes. The team has demonstrated applicability of the method in study of the effect of anti-cancer and life-prolonging drug metformin on the culture of mouse intestinal organoids.

The research has been published in high impact PLoS One journal (ranked number 25 across all disciplines, by Google Scholar Metrics in 2016).
The free full text is available here.

Comments

Post a Comment

Popular posts from this blog

Affordable microscopy of spheroid oxygenation (video protocol)

By -
Image
Our lab has published a new protocol paper, this time with the video, in the Journal of Visualized Experiments (JoVE)! In this research method paper, we report the use of red/ near-infrared emitting biocompatible nanosensors to monitor oxygenation of the live multicellular spheroids (made of one or more cell types), together with the tracers of cell death. While this methodology is typically available only for users with 'high-end' PLIM microscopes and alike, we show that using of ratiometric readout with the red and near infrared emitting fluorescent nanosensors makes this possible on broadly available 'widefield' fluorescence microscope platforms. This method helps comparing oxygenation of spheroids, assessing this in a long-term and kinetic assays, comparing their variability and viability, essential in follow-up 3D printing applications. This is the first article in our collection ' Quantitative live cell imaging of 3D models' , guest-edited together with
Read more

JoVE Methods collection: Quantitative live cell imaging of 3D models

By -
Image
Together with Prof. Margarida Barroso (Albany Medical College), we guest edit a collection of video protocols, hosted by the Journal of Visualised Experiments ( JoVE ), and  dedicated to the quantitative live cell imaging in 3D .  We expect bringing together some leaders and innovators in various aspects of the quantitative live fluorescence microscopies applied to ex vivo animal models, organoids and 3D engineered tissue blocks. The collection, which just has been started, can be assessed here .
Read more