At the time of cell culture experiments, imaging stem cells in real-time can essentially provide crucial information on cell physiology but previously, there was no such technology without the risks of phototoxicity or the use of labeling dye agents. With holotomography technology, it is possible to perform 3D live-cell imaging of the stem cells under bio-physiological conditions for real-time experiments.

Live imaging of stem cells can help in understanding the signaling and mechanisms of stem cells and primary cells in terms of growth, differentiation, and migration as real-time observation can decipher complex cell processes rather than a one-time analysis. Fluorescent imaging has been the practiced method for live monitoring but it has major limitations in terms of fluorescent tags that might interfere with cellular processes or disturb the cellular physiological environment. In case photo-sensitive molecules are present in the cells, they might cause further complications. Moreover, laser beams can damage the cells and cause phototoxicity which limits the frequency of image acquisition, if used over a long period of time. Thus, researchers have developed microscopy techniques without the use of labeling agents for sensitive cells like stem cells and primary cells.

Holotomography: an innovative technique

Scatter field data on light diffraction has been used for reconstructing 3D images by many researchers but the optical diffraction limit has prevented them often from getting a dynamic observation of cell physiology. In 2013, it was reported that YannCotte and Christian Depeursingeintroduced a new method to determine quantitative optical data of unstained living cells bypassing the limit of optical diffraction. They added a holographic innovation to the method of microscopy to live-track cells in 3D. Now, for people who do not know what a hologram is in scientific terms, holograms are3D images formed due to the interference of light beams on a sample, which is captured by a digital camera and put together by a computer after eliminating noise. This laser-based innovation in real-time microscopy, without the use of exogenous dyes, was termed as holotomography.

Benefits of Holotomography

• Low-power lasers of holotomography measure the physical properties like the refractive index, of each cell organelle, and allow the dynamic observation of cellular structures. The results arenanoscale3D real-time images of unstained living cells (Frechin et al., 2015).Using holotomography, cell compartments can be monitored at intervals of every second during growth, division, and death cellular processes.
• The absence of fluorophores eliminates the risk of phototoxicity and dismisses the possibility of interference with cellular physiology. The technology involves 100-times less power than light-sheet microscopy and laser beans are not intense.
• Live-cell imaging using holotomography can not only decipher cell processes like migration and proliferation but this 3D imaging technology can also allow the discovery of new cell processes without interfering with their normal signaling.
• Besides new cell processes, such imaging techniques can also help in detecting new therapeutic targets and help in the assessment of cellular response to new drugs.

Holotomographyis a technique that can open up several research venues only if passionate research ideas exist and researchers strive to discover newer roads to uplift biomedical and clinical research in stem cells and primary cell culture. The only challenge is to decide on how to use this innovative technology to fuel new research. For any information on innovative applications of holotomography, contact info@kosheeka.com and if you are a researcher looking forward to procuring customized primary cells and stem cells of the best quality, visit http://kosheeka.com/