Olympus IX2-RFSPOT User manual

INSTRUCTIONS
PINHOLE UNIT
AX7383
IX2-RFSPOT
The IX2-RFSPOT has been designed to be used in place of the field iris diaphragm (FS) of a IX2 Series reflected
fluorescence illuminator (IX2-RFAL). When the IX2-RFSPOT is used, a desired pinhole*can be projected on the
specimen plane, which is convenient for canceling the cage on caged specimens.
*No pinhole is provided with the unit. Please fabricate a pinhole yourself (see the figure illustrated later in this
manual) or use the commercially available recommended product (pinhole manufactured by Melles Griot, outer
diameter 16 mm).
1External View
2Precautions for Operation
· Flare is produced around the pinhole. Ghost may be noticeable when the pinhole position is deviated from the center of
the field of view.
· The pinhole image may not be able to be focused perfectly depending on the objective and wavelength in use.
· Centering of the pinhole guarantees only the movement in the center of the field of view. Movement to the peripheral
section of the field is not possible.
· The pinhole position may be deviated slightly when the pinhole is engaged or disengaged. Perform centering whenever
strict positioning is required.
· The pinhole cannot be set to an intermediate position between the IN and OUT positions.
· The IX2-SFR stage gets in the way of centering the pinhole. To prevent it, remount the IX2-SFR with the front in the back or
replace it with the IX-SVL-2.
IX2-RFSPOT
Lamp housing side
Centering screws
Pinhole engagement knob
Pushed in: Pinhole engaged (IN)
Pulled out: Vacant hole (OUT)
Clamping screw
Focus adjustment screw
Microscope frame side
Washers
(x 4)
Clamping ring

3Installation
CAUTION: To ensure safety, be sure to turn off the reflected light source before attaching.
Removing the Field Iris Diaphragm
Using the Allen screwdriver provided with the microscope frame, loosen the field iris diaphragm clamping screw on the
IX2-RFAL and pull out the field iris diaphragm to remove it.
Attaching the Pinhole
1. Using the clamping ring mounting tool provided with the pinhole unit, turn the clamping ring counterclockwise to remove.
2. Drop in the specified number of washers, determined according to the objective and excitation wavelength to be used
(see the list shown later in this manual), in the position where the clamping ring has been placed.
}If the number of washers is wrong, it will not be possible to focus on the pinhole.
3. Place the pinhole, fabricated by you or purchased commercially, above the washers, then clamp by attaching the
clamping ring again.
For the orientation of the pinhole, follow the instructions given with the figure if the pinhole is fabricated by you or insert
with the indications facing down (the lamp housing side) if the pinhole is a commercially available product.
Attaching the Pinhole Unit
Insert the pinhole unit in the microscope position where the field iris diaphragm has been, and clamp the pinhole unit
using the field iris diaphragm clamping screw.
4Operation
}Perform centering in the same way as the field iris diaphragm centering method described in the instruction manual for
the reflected fluorescence system, so that the pinhole is visible in the center of the field of view.
Focusing the Pinhole
Fit the Allen screwdriver into the focus adjustment screw and turn to adjust. (The adjustment range is ±1 mm.)
Centering the Pinhole
Insert the Allen screwdriver alternately into the two centering screws and rotate them so that the pinhole comes at the
center of the field of view.
Engaging/Disengaging the Pinhole
1. Pull out the engagement knob completely to disengage the pinhole from the light path.
2. To re-engage the pinhole in the light path, gently push the engagement knob till the deep.

Relationship Between the Number of Washers and the Objective/Wavelength (This chart is for reference only.
Depending on the objective/wavelength, the pinhole image may not be brought into focus.)
Setting the pinhole diameter
The relationship between the actual pinhole diameter and the diameter of its projection on the specimen surface is as
expressed below.
Pinhole diameter on specimen = 2X (Pinhole diameter/Objective magnification)
(Example) When the pinhole diameter is 100 µm and objective magnification is 40X:
2X (100/40) = 5 µm
#If strict pinhole diameter setting is required, actually project it on a specimen and measure the diameter using
a scale.
Pinhole Design (Unit: mm)
Pinhole position
Microscope frame side
Size indication surface of
common commercially
available pinhole
Lamp housing side
1.6
1.1
16 -0.1
-0.3
UIS2 Series
Objective Type &
Magnification
Excitation Wavelength & Priority*(Brightness/Contrast)
340 nm 360 nm 480 nm
Brightness Contrast Brightness Contrast Brightness Contrast
Number of Washers Number of Washers Number of Washers
UPlanFLN40X X X X X 1 1
UPlanSApo40X X X 0022
PlanApoN60XO X X 2122
UPlanSApo60XW X 00022
UPlanSApo60XO 000022
UPlanSApo100XO 011022
X: Unusable because of impossibility of focusing the pinhole.
* The priority can be changed between the brightness (condition in which light is gathered in the center of the field of
view) and the contrast (position where the pinhole appears sharpest although it looks a little dim) by adjustment of the
mercury lamp’s collector lens.

Printed in Japan 2004 09 M 005–@
UIS Series
Objective Type** &
Magnification
Excitation Wavelength & Priority*(Brightness/Contrast)
340 nm 360 nm 480 nm
Brightness Contrast Brightness Contrast Brightness Contrast
X: Unusable because of impossibility of focusing the pinhole.
*The priority can be changed between the brightness (condition in which light is gathered in the center of the field of
view) and the contrast (position where the pinhole appears sharpest although it looks a little dim) by adjustment of the
mercury lamp’s collector lens.
**Usable regardless of the model number (3, 2 or none).
Number of Washers Number of Washers Number of Washers
UApo20XW3/340 000022
UApo20X3/340 001122
UApo40XOI3/340 000022
UApo40XW3/340 000022
UPlanApo40X X X 0022
UPlanApo40XOI3XXXX22
PlanApo40XWLSM X X X X 2 2
PlanApo60XO3 0 X 1022
UPlanApo60XW3 X X 0022
UPlanApo60XW3/IR X X 0022
PlanApo60XWLSM X X X 4 1 2
PlanApo60XOLSM X X X X 1 2
UPlanFl60XOI3 XXXX22
UPlanApo100XOI3XX0023