Digital Staining
Figure 7 : Fixed breast cancer cell caught during division. The abnormally large nucleus (pink) and the disorganized cytoplasmic vescicles (blue-green) are highlighted in the image. Scale bar = 5 µm.
more susceptible to radiation. Gold nanoparticles refract the light in a specific way, and the 3D Cell Explorer can provide a 3D image of the cell and determine the fractions of the nanoparticles within the cell. It is not known so far where exactly these gold nanoparticles accumulate in the cell. This microscope could help in identifying which tumor cells take up the gold nanoparticles. The first results obtained with the microscope in this study are encouraging but too prelim- inary to be published. Figure 7 shows a dividing breast cancer cell provided to us by Dr. Grassberger. T e cells in this case were fi xed with 4% PFA. T e image shows the loss of internal structure typical of cancer cells, together with a large and irregular nucleus (pink). For this image the cell membrane was not visible, probably because of damage from fi xation. Internal vesicles are stained in blue and green according to slightly diff erent refractive indices. Sperm cells . A potential application is related to in vitro fertilization, a common clinical procedure with a low success rate: less the 30%. T is is mainly due to a lack of contrast on sperm cells that are selected through observation with traditional white light microscopes because conventional staining of cells for human applications is not allowed. Nanolive’s technology enables a deeper analysis of the morphology of these cells ( Figure 8 ) and thus a more accurate selection. T is may lead to an improved success rate for this routine procedure. Yeast cells . Smaller cells can be observed with the Nanolive’s technology. Figure 9 shows a few living yeast cells imaged in 1×PBS where the nucleus, vacuole, and membrane are stained in yellow, red, and blue, respectively.
Figure 8 : Comparison between (a) a classical 2D image of a fi xed sperm cell obtained with a traditional phase contrast microscope and (b) a 3D image of a fi xed sperm cell taken with the 3D Cell Explorer. The nucleus and membrane were digitally stained by the software. These images were taken at the IVF center at Hopital Cochin, Paris. Scale bar = 5 µm.
Cancer cells . The second prototype was delivered in 2015 to Harvard Medical School where the group of Dr. Clemens Grassberger is studying how gold nanoparticles make tumors
16
Figure 9 : Living yeast cells imaged with the 3D Cell Explorer. Organelles have been digitally stained to reveal the nucleus (red), vacuoles (purple), and cell membrane (blue). Scale bar = 5 µm.
www.microscopy-today.com • 2015 July
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