First Sign of Spring: Our Cells' Beauty In Bloom

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SUPPLEMENTARY FIGURE 22. Three-color multi-harmonic SI mode rendering of nuclear histones (blue), the nuclear membrane (red), and the actin cytoskeleton (green) in a fixed LLC-PK1 cell. Histones are labeled with mNeptune / H2B; the nuclear envelope is labeled with mEmerald / lamin B1; and the actin cytoskeleton is labeled with Alexa 568 / phalloidin. Scale bar: 10 micrometers.

I live in the Northeast region of the US, and around this time of year am yearning for the sight of a blossum - any sign of impending Spring. And this morning - pop! - one reveals itself in a surprising place - the March 2011 issue of {no, not Sports Illustrated, another harbinger of Spring, yes} Nature Methods.

According to Nature Methods:

Betzig and colleagues demonstrate high-speed isotropic three-dimensional imaging of living cells using a light sheet produced with a Bessel beam combined with structured illumination or two-photon excitation.

To capture these living "cells in bloom," they used an extraordinary microscope illustrated below:
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SUPPLEMENTARY FIGURE 3. Simplified schematic of the Bessel beam plane illumination microscope.
Light from laser (L) is reflected from x-axis galvanometer (XG) and transmitted in turn by relay lenses (RL) to z-axis galvanometer (ZG) and annular apodization mask (AM). XG, ZG, and AM are all at conjugate planes, so that the Gaussian beam falling on AM does not oscillate as XG and ZG are scanned. Similarly, AM is conjugate to the rear pupil plane of excitation objective (XO) so that the thin annular illumination transmitted through AM produces a Bessel beam within specimen (S) that translates along x and z without tilting. The light sheet created by scanning XG creates fluorescence at the focal plane of detection objective (DO), which is imaged at camera (C) by tube lens (TL). Different planes within S are imaged by translating DO with z-axis piezoelectric collar (ZP) in synchronization with the z axis motion of the Bessel beam provided by ZG. XO, DO and S reside in medium-filled specimen chamber (SC), and epiobjective (EO) provides a conventional view of specimen (S), for view finding purposes.

Here's a HeLa cell (human cancer cell) in all its 3D glory:

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SUPPLEMENTARY FIGURE 23. Two-color TPE sheet mode rendering of filamentous actin (orange) and connexin-43 (green) in a fixed HeLa cell. Gap junctions are labeled with mCerulean3 / connexin-43, and filamentous actin is labeled with mEmerald / Lifeact.

Want to see more? You can view 12 videos, in 3D, at the journal's website, here.

I hope this Spring brings you many beautiful blossoms. Now it's time for a nature walk.

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