18 Feb 2011,<qsg-wf1> p. 8/8
Auto Exposure
•
sets max pixels to 80% (not 100%)
(can also be applied to a selected area)
Auto Contrast
•
unlike
Gain
only
affects appearance of the image on the screen(!!!)
Keypad controls:
•
top level button 1 = transmitted light on/off (#2-4 = empty)
•
FW button switches between control of:
•
FW1 = excitation filter wheel (orbit excitation filters, only used for FRET or CFP+YFP)
•
FW2 = emission filter wheel in front of camera, underneath microscope
Æ
the orbit excitation filters must be in HOME position for use of the emission filters; if
they aren’t, press the HOME button on the external keyboard
Control buttons on the lefthand side of the microscope:
•
lowest button
:
•
switches between frontport (empty) <> eyepiece <> baseport (camera)
•
middle button
:
•
switches between sideport 100% (empty) <> 50/50 <> eye/ front/ base 100%
•
top:
Optovar in / out
File formats
•
The original Volocity libraries are stored as "mvd2" files together with a folder containing the
image data in "aisf" files
•
TIFF
:
looses all enhancements but keeps original ("raw") data
•
all stacks/ channels are stored in 1 multipage TIFF with the original bit-depth (8 or 12 bit)
•
TIFF for publication
•
screen version of image, including colours, merge, enhancements, on-screen image
resolution, it is always 8-bit
Æ
loss of original ("raw") data!!!
merge colours:
•
CREATE NEW IMAGE SEQUENCE > MULTIPLE CHANNELS
•
drag single channel (or z / timepoint) images onto new image sequence window
