Olympus FV300 User Manual

CONFOCAL LASER SCANNING

BIOLOGICAL MICROSCOPES

FV300

FLUOVIEW

1

FV300 CONFOCAL LASER SCANNING MICROSCO
PERFORMANCE FOR THE PERSONAL USER

The FV300 is the ideal choice of laser scanning microscopes for personal users.
Its optical system is fully integrated, from scanner to microscope, and not only delivers
outstanding optical sectioning, but also ensures the easy, flexible expandability required
for any future upgrade.

With its wide choice of options and configurations, including the Olympus inverted,
upright and fixed-stage upright microscope platforms, the FV300 offers excellent
versatility as well as top-class laser scanning performance.

• Highest image quality (12 bit, 2048x2048 pixel resolution)
with economical cost

• Easy operation, with user-friendly software

• Simultaneous capture of 2 fluorescence and 1 transmitted
light detector images

• Capable of the most demanding tasks, with a direct and
efficient optical system

• Optical system chromatically corrects aberrations from UV
to NIR (near infrared red) spectrum

• Fiber illumination system separates fluorescence and
transmitted light sources from the microscope body for
improved temperature stability

FV300-IX71configuration

2

q

e

r

t

y

u

i

o

!0

!1

!2

w

q Optical fiber for laser introduction
w Beam collimator
e Polarizer
r Dichromatic mirror
t Excitation dichromatic mirror
y XY galvanometer mirror scanners
u Pupil lens
i Collector Lens
o Pinhole turret
!0 Emission beam splitter slider
!1 Barrier filter slider
!2 Photo multiplier

Preset the conditions for image
acquisition and loading

Storage of Acquisition Settings enables
immediate, one-touch recall of all the
relevant experimental settings and
conditions. Adding new conditions or
altering existing ones is quick and easy.

AOTF: flexible control of the laser
intensity to meet the specific
demands (optional)

The laser exposure will be limited within
the scanning area by default, minimizing
unnecessary bleaching of the specimen.
Option includes:

•Any laser intensity for any excitation area
("Region of Excitation" mode)

•Multiple laser applications

•AOTF controller that provides easy link
with external equipment

ZoomIn scanning

Zoom scanning can be
conducted over any
designated rectangular area.

Since only the region of the
targeted, zoomed-in area is acquired,
scan time and laser exposure of the
specimen is minimized.

Point scanning

The ultimate in fast scanning,

the point scan enables

accurate quantitation of

intensity changes during rapid

physiological events.

Line scanning

A single line may be scanned,
oriented at any angle in the XY
plane. This fast scanning

option permits accurate
quantitation of physiological events such
as Calcium waves or sparks.

Free line scanning

Intensity changes may be
measured over a given period
of time along the length of a

freely drawn line, such as the
trace of an axon or along a cellular
junction.

Clip scanning

By cropping the image, selected areas
can be cut out of complex image stacks.

3

Flexible setting of scanning size,
zoom, movement and rotation

The observation field and scanning area
are both displayed graphically. Settings
can be confirmed while scrolling through
the zoom ratios. The “pan” button lets the
operator move the image acquisition area
at will, and rotation scanning of images is
also possible.

Simplified toolbar

A newly designed toolbar with various
dedicated buttons has greatly improved
ease of use. The user can execute a
succession of selected processes with
one-click operation for each.

Ultimate ease of operation and monitor display.

Software Graphical User Interface

Dye selection display

When a fluorescence dye is chosen, the
laser and light path settings are selected
automatically, with each of the selected
fluorescence dyes displayed graphically
on the monitor.

Innovative scanning method for
improved performance

ZoomIn

Spot

SlantLine

FreeLine

20%

50%

B

Versatile display options

Exchange between condensed and full
image display modes can be performed
with a single touch. Individual panel
layouts can be changed at will, and the
panel in use can be placed in any desired
position.

Image tool bar

X-Y-Z scanning operations and time­lapse observations both produce multiple
images, which can be displayed in
sequence simply by clicking the
sequential mode button. Channel
selection and image zooming are also
available on the same menu.

4

Thumbnail display

Data stored in the gallery window are
displayed as thumbnails for easy viewing.

Sequential scanning to prevent cross talk

Sequential scanning may be used to minimize the fluorescence cross talk
often seen between channels in multicolored samples. This is achieved by
exciting each fluorochrome independently, one dye at a time. With the AOTF
function, line sequential scanning is available as well.

* Once optimized: steps q-r can be performed easily

Scanning unit set-up monitor display

Human colon crypt

Nuclei (Blue) TO-PRO-3
Actin (Green) Alexa 488
APC gene product (Red) Alexa 568

Christine Anderson,
Laboratory of Prof. Ray White,
Hunstsman Cancer Institute, Utah

q Excited by only HeNe633

w Excited by only Kr568

e Excited by only Ar488

r Composition

Easy exchange between display
methods

Independent navigation bars for each
image window enable the display method
to be changed quickly and easily.

Tiling display function for see-at-a­glance comparison of multiple images

The FV300's live tiling function, which is
especially valuable in time course
experiments, allows observations of
multiple images or changes in the
specimen while the experiment is in
progress. Images in series (e.g. XYt or
XYZ) can be freely displayed.

Single monitor display is also possible.