Production and Multi-Parameter Live Cell Fluorescence Lifetime Imaging Microscopy (FLIM) of Multicellular Spheroids (JOVE protocol)

By -


ALERT: New publication from our group!

Angela, Gabriele, Hang, Nore, Nina and Irina teamed up to produce a video protocol paper, focusing on comparing various multicellular spheroid formation methods, suitable for 'downstream' live multi-parameter fluorescence lifetime imaging microscopy (FLIM) and imaging cell oxygenation. This protocol is illustrated with examples of autofluorescence-based FLIM microscopy of NAD(P)H, oxygenation and cell death, as well as imaging of visible flavin-based cell autofluorescence, performed in tumour and stem cell-derived spheroids. The article is also a part of the Methods collection 'Quantitative Live Cell Imaging of 3D Models', which is going to expand in the future. 


 

Link to the publication:

https://www.jove.com/t/66845/production-multi-parameter-live-cell-fluorescence-lifetime-imaging

The Methods Collection (Journal of Visualised Experiments, JOVE): https://app.jove.com/methods-collections/902


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