RI segmentation (D-stains)

On this page you will have the possibility to explore a new way to analyze your data: RI segmentation (D-stain)! Enjoy and stay tuned: the list of results grows every week…

Nanolive’s technology allows you to:

  • Differentiate sub cellular populations in co-colture
  • Use the chromatin RI as a marker of each mitotic phase
  • Discriminate different microorganism strains like yeast and bacteria

Segmentation of intact cells is a critical task in automated image analysis and quantification of cellular microscopic images. Automatic segmentation in 3D microscopy images is essential for many biological studies, including high throughput analysis of gene expression level, morphology, and phenotypes at single cell level. Our digital staining panel allows us to discriminate different cell types and cellular components based only on their specific refractive index (RI) range. The D-stain remains robust and consistent among several acquisitions and during the whole real-time acquisition, allowing an automatic segmentation of cellular populations.  

Monitoring of DNA refractive index changes during mitosis 

Fibroblastic Reticular Cells (FRCs) were grown to 60% confluency in complete DMEM medium (Dulbecco’s Modified Eagle Medium) in 35mm glass bottom culture dishes (FluoroDishes™ WPI, #FD35-100). The time-lapse imaging experiment was conducted with a standard top-stage incubator set to 37°C and 5% CO2 for 45 minutes, capturing images every 30 seconds.

The refractive index of the DNA changes during mitosis. The 3D Cell Explorer is able to discriminate different mitotic phases based on these chromatin refractive index changes. 

Co-culture: Melanoma cancer cells and amoeba cells 

Mouse skin melanoma cancer cells (B16, p35) were grown to 40% confluency in complete DMEM medium (Dulbecco’s Modified Eagle Medium) in 35mm glass bottom culture dishes (FluoroDishes™ WPI, #FD35-100). They were incubated overnight with dictyostelium amoebae cells (WT1, p14) grown in HL-5 medium. The time-lapse imaging experiment was conducted with a standard top-stage incubator set to 37°C and 5% CO2 for 8 hours, capturing images every minute. Our digital staining panel allows us to discriminate mammalian (blue and violet) and amoeba cells (green) based on their specific refractive index (RI) range. The d-stain remains robust and consistent among several acquisitions and during the whole time-lapse (8h) allowing an automatic segmentation of cell populations.

Bacteria and Yeast automatic segmentation 

Fibroblastic Reticular Cells (FRCs) were grown to 60% confluency in complete DMEM medium (Dulbecco’s Modified Eagle Medium) in 35mm glass bottom culture dishes (FluoroDishes™ WPI, #FD35-100) and fixed with PFA 4% (15min). Then the cell culture was contaminated by wild bacteria and yeast strains. Mammalian cells (big, flat, purple borders) and the other biological contaminants can be easily distinguished by shape and dimensions. More than that, our digital staining panel allows us to make a segmentation of the different yeast stains (yellow and blue) based on their specific refractive index (RI) range.

Learn more about the 3D Cell Explorer

Read more about the 3D Cell Explorer here and learn how to use it in 1 minute with this video

Discover all the secrets of the 3D Cell Explorer on our Introductory webinar

Learn how to digitally stain your cells with our software STEVE, in 1 minute

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Related topics

Read our successful blog post about “Cancer cells and amoeba in co-culture

Automatically segment yeast and bacteria with the 3D Cell Explorer!

Check out our blog all about segmenting Circulating Tumor Cells in the blood!

Did you know that the chromatin’s refractive index changes depending on its cell cycle phase? 

Find our latest publications here