Biology Reference
In-Depth Information
FIGURE 22.4
Preparation of microscope slides with agarose pads. See text for details.
8 s. With these conditions, time lapses can be captured for as long as 8 min without
significant photobleaching. Although precise measurement of microtubule lengths
will be somewhat compromised, time lapses can be extended by reducing excitation
intensity or exposure time and binning images 2
<
2.
Microtubule length can be measured using commercial (e.g., Slidebook/
Metamorph) or public domain (ImageJ) software. Over subsequent time points, the
lengths of individual microtubules are determined from the base of the microtubule,
where they anchor to the spindle pole body, to the opposite tip of the microtubule.
The measurement should accommodate the three-dimensional length by utilizing
the z-focal planes. Care should be taken that z-series encompass the whole cell,
