Time Series
Dynamic processes in living specimens can be
recorded by means of time series. Data thus
acquired can be analyzed “off-line”, i.e. after image
acquisition, or “on-line”, i.e. right during the experi-
ment, for example in the Online Ratio mode.
Time series are defined by a start time and the
time interval between two successive images. The
series can be started by a mouse click, automati-
cally at a preselected time, or by some external
trigger. To analyze a time series, the Physiology
software option allows fluorescence intensity
changes to be quantified in defined regions of
interest (ROIs).
Within a time series, the LSM 510 permits selec-
tive, point-accurate illumination of ROIs with laser
light.
This function is useful for generating a photo-
bleaching routine within a FRAP experiment
(fluorescence recovery after photobleaching), for
analyzing dynamic processes, and for the photo-
activation or photoconversion of suitable fluo-
rochromes. Complex time series experiments, with
different images to be taken at different sites
within a specimen according to a defined time
pattern, can be defined by means of a special soft-
ware option.
The procedure for a time series
1
Define the image dimensions to be recorded versus time
(XY
image,
Z stack,
λ
stack)
2
Optimize the recording conditions at minimum laser output to avoid or minimize bleaching
3
Define the number of images to be taken and the time interval between
two successive images
(Time Interval
or
Time Delay
)
4
Combine with a photobleaching routine if required: define the region to be bleached,
the laser line and its power, the number of bleaching actions,
and the bleaching start time within the series
5
Start the time series with the Start button, at a preselected time, or by an external trigger
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