Chapter 2 Microscopy
22
2.4 Episcopic Fluorescence (Epi-FL) Microscopy
1
Configure the microscope to the basic
state.
1-1 Turn on the power to the system.
[a] Power switch (
☞
3.1)
▼
1-2 Place the 10x objective into the optical path.
[b]
Nosepiece
(
☞
3.9.1)
▼
1-3 Switch the optical path of the tube base unit
to
(binocular part).
[c] Tube base optical path changeover knob
(
☞
3.7.1)
▼
1-4 Switch the optical path to the required
observation port.
[d] Optical path changeover dial (
☞
3.2.1)
▼
1-5 Configure the following controls to the basic
state.
[e] Intermediate magnification dial
(Set to 1x:
☞
3.2.2)
[f] Objective with a correction collar
(Set the correction collar to the reference position:
☞
3.10.1)
▼
1-6 Remove unnecessary optical elements from
the optical path.
• Analyzer slider (
☞
3.5.2)
• DIC slider (
☞
3.9.2)
Configuration of the microscope
For epi-fl microscopy, change the illumination to the epi
illumination, arbitrarily select a filter cube, and place it into
the optical path.
Note: Letters within [ ] correspond to those in the procedure.
2
Change illumination to epi, set a sample, and
focus on it.
2-1 To prevent the dia-illumination LED from
emitting autofluorescence due to
epi-illumination, close the dia-illumination
shutter.
[g] Dia-illumination shutter knob (
☞
3.3.1)
[h] Condenser turret (
☞
3.4.6)
▼
2-2 Specify the FL turret address and place the
filter cube suitable for the sample into the
optical path.
[i] FL turret (
☞
3.12.1)
▼
2-3 Fully open the epi aperture diaphragm.
[j] Aperture diaphragm open/close lever (
☞
3.13.1)
▼
2-4 Place the field diaphragm slider into the
optical path, and fully open the epi field
diaphragm.
[k] Field diaphragm slider (
☞
3.13.3)
[k] Field diaphragm open/close lever (
☞
3.13.3)
▼
2-5 Place the sample on the stage.
[l]
Stage
(
☞
3.8.1)
When placing a large sample on the stage, tilt the pillar
for dia-illumination backward to obtain sufficient space.
(
☞
3.3.5)
▼
2-6 Open the FL turret shutter.
[m] FL turret shutter knob (
☞
3.12.2)
The epi illumination reaches the observation port.
▼
2-7 Turn on the epi illumination and adjust the
brightness.
[n] Light source for episcopic illumination (
☞
3.14.1)
▼
2-8 Move the required part of the sample into the
center of the field of view.
[o] Stage handle (
☞
3.8.3)
▼
2-9 Darken the room.
When the contrast shield is used, close it to shield light
from the above.
(
☞
3.3.6)
▼
2-10 Focus on the sample.
[p] Focus knob (
☞
3.11.1)
▼
2-11 Adjust the diopters and the interpupillary
distance if required.
[q]
Eyepiece
(
☞
3.6.2)
[r] Binocular part (
☞
3.6.3)
[a]
[b]
[c]
[l]
[g]
[p]
[q]
[e]
[k]
[n]
[f]
[r]
[d]
[i]
[o]
[j]
[h]
[m]