Stem Cell ResearchStem cell research with label-free 3D live cell imaging
Advancing stem cell research with the 3D Cell Explorer
Scientists can discover more about the behavior of stem cells with Nanolive’s 3D live cell microscope – the 3D Cell Explorer. This microscope offers the best contrast, resolution and versatility that a label-free microscopic approach can offer today, coupled to unmatched time resolution. Last but not least, it’s 3D.
Please have a look at our stunning material: stem cells as never seen before!
Long Term Live Imaging of Mouse Embryonic Stem Cells for 15 hours
Long Term Live Imaging of Mouse Embryonic Stem Cells for 34 hours
Spectacular Mitosis in Mesenchymal Stem Cells
Mesenchymal stem cells are multipotent cells that can be obtained from umbilical cord tissue, adipose tissue1, dental pulp or amniotic liquid. The principal source of mesenchymal stem cells is the human bone marrow, which constitutes 4% of the total body mass of an individual2. They are able to differentiate to a variety of mesenchymal tissues lineages such as cartilage, fat, bone, muscle, tendon, and stromal tissue2.
This capacity to nourish tissue regeneration have positioned MSCs as promising medical treatments, with some studies already giving results in their applications in inflammatory bowel disease3 and other immune disorders4, or in ischemic heart disease5.
The initial enthusiasm, though, has been partly shadowed by the lack of a standardised and well-detailed cell processing and culture conditions protocols, which led to repeatability and scaling problems6.
Besides, mesenchymal stem cells are sensitive to experiment induced stresses such as phototoxicity or bleaching, present in fluorescence microscopy, the current method of choice for stem cells imaging. These types of stresses lead to a limitation in the cultured cells imaging possibilities. Implementation of the use of the 3D Cell Explorer microscope would help avoid these perturbations and improve this fundamental research as the samples need no preparation, which allows for a fast, non-invasive and expertise-independent live observation of mesenchymal stem cells. In addition, the 3D Cell Explorer laser uses 100 times less energy than the least energetic laser in the current fluorescent imaging approaches, which makes long-term imaging (up to weeks) possible.
In the provided example of a spectacular cell division taking place in a living sample of human mesenchymal stem cells kindly provided by Promocell GmbH cultured with low-serum cell growth medium7 and observed under the 3D Cell Explorer. The characterization of the different steps and structures of mitosis was possible.
High-frequency, long-term live imaging
One of the biggest challenges in cell biology is long-term imaging of fine cellular dynamics. In particular, the 3D Cell Explorer generates no phototoxicity as it injects in the sample ~100 times less energy (~0.2 nW/µm2) than light sheet microscopes (~1nW/µm2) that are the reference in the matter. With a resolution of 195nm (higher than standard confocal) it enables high resolution and high-frequency imaging even with sensitive material, such as stem cells*.
Unique dynamics of cellular organelles or highly dynamic and small cellular structures can now be clearly analyzed, such as mitochondria, lipid droplets, filopodia, dendrites, axons, nucleus and nucleoli etc.
With Nanolive’s live cell imaging tool, it is possible to perform endless live imaging at the maximal acquisition speed (1 image per 1.7 seconds).
This certainly exceeds any live cell imaging capabilities of any other microscope and will revolutionize live cell imaging of stem cells.
(*plus, the next generation holotomographic technology used to make these movies is sold at a fraction of the cost of a standard confocal)
Long Term Live Imaging of Mouse Embryonic Stem Cells for 34 hours!
Long Term Live Imaging of Mouse Embryonic Stem Cells for 48 hours! Part 2
Watch our Webinar: Long-term 3D imaging and quantification of stem cell dynamics
Dr. Mathieu Frechin, Head of Quantitative Biology at Nanolive introduces you to our holotomographic microscopy and its implications in long-term imaging of stem cells. He outlines the advantages that Nanolive’s holotomographic technology has to study sensitive cell lines. In particular, the following topics are being covered:
- Challenges in live cell imaging of stem cells
- State of the art of label-free microscopy for stem cells
- Highlights of our holotomographic approach on stem cell research presenting movies and interesting biological dynamics.
Stem Cell Research with the 3D Cell Explorer-fluo
Live FUCCI mESC imaged for over 48 hours
The video shows live FUCCI mESC that were imaged for over 48 hours with Nanolive’s 3D Cell Explorer-fluo. A holotomographic image was taken every 14 seconds while a double channel (green and red) epifluorescence image was only taken every 30 holotomographic frames (7:30 minutes). The difference in acquisition rate was performed to reduce fluorescence related phototoxicity.
*Please note that the latest fluorescence image is displayed over the following 30 holotomographic images until a new one is acquired, for visualization purpose. The mismatch between the two signals is due to this image processing choice.
The 3D Cell Explorer-fluo
Multimodal complete solution for 3D live cell exploration
Combine 3D refractive index analysis with a fully integrated 3 channel fluorescence module to image your live cells as they are and as long as you want. Put chemical information into structural context for new biological insights.
Transform 2D fluorescence into 3D cell tomography: Identify cell organelles through fluorescence and monitor non-invasively their structures and dynamics in 3D and marker-free. Explore fluorescence (3 channels) and Digital Stains (8 channels) simultaneously.
Extended live cell imaging
Image your live cells as long as you need. Limit cell damages caused by fluorescent markers, bleaching and phototoxicity.
Application Note: Growing and Filming Stem Cells with the 3D Cell Explorer
This application note focuses on the proper set up of the 3D Cell Explorer with its incubation system to perform long-time imaging (up to weeks) of mammalian stem cells, at high spatio-temporal resolution and in 3D.
- Bunnell, B. A., Flaat, M., Gagliardi, C., Patel, B. & Ripoll, C. Adipose-derived stem cells: Isolation, expansion and differentiation. Methods 45, 115–120 (2008).
- Pittenger, M. F. et al. Multilineage potential of adult human mesenchymal stem cells. Science 284, 143–7 (1999).
- Research Supports Promise of Cell Therapy for Bowel Disease | Wake Forest Baptist Medical Center. Available at: https://newsroom.wakehealth.edu/News-Releases/2013/02/Research-Supports-Promise-of-Cell-Therapy-for-Bowel-Disease. (Accessed: 1st May 2019)
- Wang, M., Yuan, Q. & Xie, L. Mesenchymal Stem Cell-Based Immunomodulation: Properties and Clinical Application. Stem Cells Int. 2018, 1–12 (2018).
- Ward, M. R., Abadeh, A. & Connelly, K. A. Concise Review: Rational Use of Mesenchymal Stem Cells in the Treatment of Ischemic Heart Disease. Stem Cells Transl. Med. 7, 543–550 (2018).
- Robinson, P. G. et al. Reporting of Mesenchymal Stem Cell Preparation Protocols and Composition: A Systematic Review of the Clinical Orthopaedic Literature. Am. J. Sports Med. 47, 991–1000 (2019).
- Mesenchymal Stem Cell Growth Medium 2 | PromoCell. Available at: https://www.promocell.com/product/mesenchymal-stem-cell-growth-medium-2/. (Accessed: 6th May 2019)