Ever wondered how to measure cell volume with high precision? Ask STEVE!

Cell volume is a critically fundamental parameter in biology and medicine. Optical technology offers many possibilities for cytometry, but obtaining an accurate optical measurement of cell volume is still very challenging. The most used method so far is indirect measurement, where the cell is submerged in a fluid-filled container and the displaced volume is estimated. 

Nanolive’s 3D Cell Explorer goes beyond these limits and allows for direct volume measurements of biological samples. In this post we will show you how to monitor, in real-time, the cell volume in a precise and quantitative way using the 3D Cell Explorer and its software STEVE!

How does it work? 

  1. Digitally stain (D-stain) your region of interest (in our case the whole cell)
  2. Click on the “i” button under the selected stain color
  3. Obtain in this way the total volume (μm3) covered by that specific D-stain (Fig.1). This corresponds to the exact volume of your cell!
Fig.1

Figure 1. How to obtain volume measurement through STEVE

 

The accuracy of the tool was tested by using Sigma-Aldrich micro-particles based on silicon dioxide (44054): based on datasheet, the average bead diameter is of 4.8±0,15 μm and which correspond to 52,6 – 63,5 μm3 in volume range. The beads volume measured with STEVE resulted in 56μm3, falling within the estimated measurement.

Figure 2. Sphere measurement. Four silica spheres with a known volume were measured as a control. The average diameter size was of 4.7±0.4.

 

We applied this tool to evaluate the volume of a variety of biological samples.

From the big to small:

A big eukaryotic protozoa, Colpidium: 

colpidum plus

Figure 3. Colpidium colpoda cell. The total cell volume was digitally stained in red with STEVE. On the left is shown the D-stain panel and on the right the 3D reconstruction of the whole cell volume. The volume of the D-stained pixels give us the total cell volume.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Mammalian skin cancer cell:

cell plus

Figure 4. Mammalian skin cancer cell. The total cell volume was digitally stained in red with STEVE. On the left is shown the D-stain panel and on the right the 3D reconstruction of the whole cell volume. The volume of the D-stained pixels give us the total cell volume.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Human sperm cell:

sperm plus

Figure 5. Human sperm cell. The total cell volume was digitally stained in red with STEVE. On the left is shown the D-stain panel and on the right the 3D reconstruction of the whole cell volume. The volume of the D-stained pixels give us the total cell volume.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Lactobacillus (bacteria cell):

bacteria plus

Figure 6. Lactobacillus (bacteria). The total cell volume was digitally stained in red with STEVE. On the left is shown the D-stain panel and on the right the 3D reconstruction of the whole cell volume. The volume of the D-stained pixels give us the total cell volume. The total volume was following divided by four since there are four cells in the field of view.

 

 

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