Long-Term Recordings of Live Human Cells Using Phase Contrast Microscopy - Biomedical Optical Phase Microscopy and Nanoscopy - page 50

stage to make enough distance for the scribe, and rotate the nosepiece so that the scribe
is centered over the field of interest. Select the diameter of the circle by adjusting the
rotatable ring on the barrel of the scribe. While watching from the side, carefully raise
the stage until the coverslip lightly contacts the diamond tip of the scribe. Gently rotate
the knurled ring on the scribe to rotate the diamond tip. Light contact is sufficient to
make an easily visible scribe groove without cracking the coverslip. After scribing,
lower the stage away from the tip of the scribe and rotate in an objective. Before taking
the preparation off the microscope it is important to take an image of the cells in
relation to the slightly out of focus scribed circle ( Figure 3.2 ). This image can be
printed and will serve as a map to identify and relocate cells of interest after fixation
and immunostaining.
2. Once filming is terminated, the cell bearing coverslip is carefully removed (see protocol
#8) and immediately fixed in whatever manner one is using.

References

[1] Y. Uetake, J. Loncarek, J.J. Nordberg, C.N. English, S. Laterra, A. Khodjakov, et al., Cell cycle
progression and de novo centriole assembly after centrosomal removal in untransformed human cells,
J. Cell Biol. 176 (2007) 173 182.
[2] C.L. Rieder, R.W. Cole, Cold shock and the mammalian cell cycle, Cell Cycle 1 (2002) 169 175.
[3] P.N. Rao, J. Engelberg, Hela cells: effects of temperature on the life cycle, Science 148 (1965)
1092 1093.
[4] G. Sluder, J.J. Nordberg, F.J. Miller, E.H. Hinchcliffe, A sealed preparation for long-term observations of
cultured cells, in: D.L. Spector, R.D. Goldman (Eds.), Live Cell Imaging: A Laboratory Manual, Cold
Spring Harbor Laboratory Press, Cold Spring Harbor, 2005, pp. 345 349.

Further Reading

http://micro.magnet.fsu.edu/primer/techniques/livecellimaging/index.html —Introduction to Live-Cell Imaging
Techniques by Florida State University.
http://www.microscopyu.com/articles/livecellimaging/index.html —Introduction to Live-Cell Imaging
Techniques by Nikon
http://www.olympusmicro.com/primer/resources/livecells.html —Live-Cell Imaging Resources by Olympus
J.C. Waters, Live cell fluorescence imaging, in: G. Sluder, D.E. Wolf (Eds.). Digital Microscopy, third ed.,
Methods in Cell Biology 81 (2007) 115 140, Elsevier, Academic Press, London.