3D live and label-free cells: Mouse Ebryonic Stem cells (mESCs) mitosis.

[Evi_lonan]

Protocol:
The cells were grown to 60% confluency in complete DMEM medium in ibidi 35mm glass bottom dish. The time-lapse imaging experiment was conducted with a ibidi-nanolive top stage incubator tor set to 37°C and 5% CO2 for 8h, capturing images every 30 seconds.
The image was taken using Nanolive's technology: 3D Cell Explorer. A label-free holo-tomographic microscope able to reconstruct in 3D living cells based on their optical density.

Image description:
Nanolive digital staining allows the discrimination of chomosomes (yellow) and cell membranes (blue) based on their specific refractive index (RI) range.

[IanW]

This is astounding to look at. Is there any chance that you could explain the process - but aimed at one who is not all that well versed in the science and technology you speak of? Thanks for the posting of this.

[wporter]

What would help your exposition regarding this new and impressive technology (not to mention your marketing), is a few side-by-side images, comparing the same subject using the best of normal technology, to those taken with your new microscope, so people can see the improvement from the new tech. Otherwise, they're just as likely to say to themselves, "Well, can't that be already be done with [insert older technique here]?"

[mrsonchus]

Hmm, I was thinking, based upon these particular images, exactly that wporter old chap.

The description sounds a lot like a 'coloured' phase contrast, whilst the images are, to my eyes, suggestive of fluorescent-label imaging...

As suggested, direct-comparison would be a great aid to evaluation I think.

John B.

sounds good - looks familiar

[Evi_lonan]

This is astounding to look at. Is there any chance that you could explain the process - but aimed at one who is not all that well versed in the science and technology you speak of? Thanks for the posting of this.

In this video (here image) you can observe a cell division. During the cell division, the chromatin (the carrier of the genetic information) condenses and you can see how the chromosomes (displayed in yellow) are distributed equally to the two daughter cells. With our new revolutionary microscope, we are able to measure and visualize the refractive index of cells and subcellular structures – such as the chromosmes visualized here – and observe them in living cells over long time periods. All these images were recorded without any staining. It is possible to just color the structures according to their refractive index.

[Evi_lonan]

What would help your exposition regarding this new and impressive technology (not to mention your marketing), is a few side-by-side images, comparing the same subject using the best of normal technology, to those taken with your new microscope, so people can see the improvement from the new tech. Otherwise, they're just as likely to say to themselves, "Well, can't that be already be done with [insert older technique here]?"

Thank you very much for your input. You are absolutely right, it would be a nice addition to our images. If for example people could see our 3D images in contrast to the conventional 3D DIC images, or directly see the enhanced resolution compared to standard fluorescence images, as well as the reduced cytotoxicity compared to regular confocal approaches in side to side images.

[Evi_lonan]

Hmm, I was thinking, based upon these particular images, exactly that wporter old chap.

The description sounds a lot like a 'coloured' phase contrast, whilst the images are, to my eyes, suggestive of fluorescent-label imaging...

As suggested, direct-comparison would be a great aid to evaluation I think.

John B.

sounds good - looks familiar

Indeed you see images of unstained cells here. With the 3D Cell Explorer it is possible to pseudo-color the images according to the refractive indices of the structures. In this case the refractive index of the chromosomes is pseudo-colored in yellow, while the cell membrane is displayed in turquois and the cytoplasm in violet. And the whole process can be observed in 3 dimensions over time.