Application of the live-cell imager Holomonitor M4 in holographic dynamic monitoring of organoid growth and migration

Organoids refer to tissue analogues

However, there are many challenges in filming and quantifying organoid cultures:

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Holomonitor M4's powerful holographic dynamic monitoring and analysis functions help you easily solve the pain points in the process of organoid shooting, and the small body of Holomonitor M4 can monitor organoids in the incubator for a long time without labels to avoid environmental changes; Multiple free selection points can be performed for different well plates to ensure experimental reproducibility, and up to 96 well plates can be monitored; 3D imaging and powerful analysis functions can not only provide information on morphological changes in organoid areas, but also provide rich information

Here's a case

Oropharyngeal squamous cell carcinoma (OPSCC) stem cells have the ability to self-renew and differentiate into tumor heterogeneous three-dimensional structures

Monitor the number of organoids formed to assess the optimal inoculation density

Highlights: The Holomonitor M4's small body allows long-term label-free real-time monitoring of organoids in the incubator to avoid environmental changes

In this paper, the time for cells to aggregate to form organoids at different seed densities was observed continuously for 15 hours, and the overall growth status was

Figure 1: Determination of inoculation density conditions and optical properties

Long-term monitoring and real-time quantification of organoid growth status

Highlights: HoloMonitor M4 3D imaging and powerful analysis capabilities to quantify organoid area and volume changes in real time

This paper evaluates the events affecting organoid formation and growth through 15-hour linked observations, and the results show that organoids begin to form 2 h after inoculation, and monitor overall growth

Figure 2: Monitor organoid formation over a 15-hour period using M4

Analysis of the ability of different genotypes of cells to form organoids and their movements

Highlights: HoloMonitor M4 can be freely selected for different well plates, which can ensure experimental reproducibility; Powerful analytical functions provide rich information
such as cell migration distance and speed.

During OPSCC organoid formation, 14% of the cells did not form any 3D structure, indicating that there may be subpopulations
of cells with different organoid-forming abilities.
It is assessed
by classifying organoid capacity and motility differences based on CD44 and NGFR (expression) for individual OPSCC cells.
Cells of three different phenotypes, CD44+/NGFR+, CD44+/NGFR−, and CD44−/NGFR−, were inoculated in a hydrogel and tracked
using HoloMonitor M4.
The results showed that CD44+ expressed cells could form organoids within 15 h and showed greater mean motility (140-240 μm), while CD44− tumor cells were unable to form organoids and moved slowly in hydrogels (~30 μm).

Figure 3: Organoid capacity and motility of different oropharyngeal squamous cell carcinoma-cellular phenotypes
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(b) Average cumulative motility of individual cells is obtained based on the expression of CD44 and NGFR, with repeated monitoring
of organoids of five phenotypes.

References:

[1] Gomez Jimenez D, Carreira Santos S, Greiff L, et al.
Subpopulations of Organoid-Forming Cells Have Different Motility.
Applied Sciences.
2020; 10(13):4673.

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