Leica DMIRE2 Manuel utilisateur
CMN Imaging Core
January 16, 2013
1
LEICA DM IRE2 MICROSCOPE MANUAL
Neuroscience Imaging Core
Rightmire Hall
Ohio State University
Director: Tony Brown
Rightmire 060
292-1205
Facility Manager: Paula Monsma
Rightmire 062
292-3025
This manual prepared by Tony Brown.
CMN Imaging Core
January 16, 2013
2
TABLE OF CONTENTS
LEICA DM IRE2 MICROSCOPE MANUAL................................ 1
Getting to know the microscope.............................................. 3
Microscope control panel......................................................... 7
LCD display panel...................................................................... 8
LCD display panel buttons ....................................................... 9
Changing filters .......................................................................10
Changing objectives................................................................ 11
To switch between dry and immersion objectives............... 12
Adjust halogen lamp brightness............................................13
To focus using the focusing knobs.......................................14
Using the focusing buttons....................................................15
Changing the coarseness of the focusing............................16
Eyepieces ................................................................................. 17
Placing a slide on the Z-galvo stage...................................... 17
Transmitted light detector selection knob............................ 18
Bright field observation .......................................................... 18
Koehler illumination ................................................................19
Phase contrast......................................................................... 20
Differential interference contrast (DIC) .................................21
Switching stage holders .........................................................22
Using immersion objectives...................................................23
Cleaning objectives................................................................. 24
Applying oil to objective without removing slide.................25
Specimen preparation............................................................. 26
DOs and DON’Ts......................................................................27
CMN Imaging Core
January 16, 2013
3
Getting to know the microscope
VIEW FROM LEFT
Microscope
control panel
Laser scan
head
with 3
fluorescence
detectors
Condenser
lens
Side port
Focus knob
Eyepieces
Transmitted
light detector
Z- galvo stage
(detachable)
Halogen lamp
intensity
adjustment
wheel
Filter cube
turret
Transmitted light
illumination column
(tilts back for better
access to stage)
CMN Imaging Core
January 16, 2013
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VIEW FROM RIGHT
Halogen lamp
housing
(transmitted light)
Mercury lamp
housing
(epi-fluorescence)
Focus knob
X-Y stage
movement
Filter holders
(empty)
Transmitted light
detector
selection knob
Condenser turret
Tube lens module
with Bertrand lens
Objective turret
CMN Imaging Core
January 16, 2013
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OBJECTIVE TURRET (DETAILED VIEW)
objective
turret
objective
prism turret
filter cube
turret
cuvesfou
DIC analyzer
Epi-fluorescence
field diaphragm
(controls area of
illumination)
Epifluorescence
Illumination
diaphragm
(controls
brightness of
illumination)
CMN Imaging Core
January 16, 2013
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OBJECTIVE PRISM TURRET POSITIONS
Objective
turret
position
Description
Matching objectives
BF
Empty (for bright field
or phase contrast)
C
DIC objective prism
20x multi-immersion
D
DIC objective prism
63x water
immersion
100x oil immersion
E
DIC objective prism
40x oil immersion
63x oil immersion
CONDENSER TURRET POSITIONS
Condenser
turret
position
Description
Matching objectives
BF
Empty (for bright
field)
PH1
phase ring
10x dry
PH2
phase ring
40x dry
20
DIC condenser
prism
20x mulit-immersion
40/63
DIC condenser
prism
40x oil immersion
63x water immersion
63/100
DIC condenser
prism
63x oil immersion
100x oil immersion
CMN Imaging Core
January 16, 2013
7
Microscope control panel
LCD display
switch filter cube
GFP: green fluorochromes
RED: red fluorochromes
CFP/: far red fluorochromes
SCAN=empty slot
select port
VIS=light to eyepieces
SIDE=light to scanner
BOTTOM: disabled
switch tube lens
(disabled)
LCD display
panel buttons
Red light
shows current
filter selection
Mercury lamp
shutter
(red light is on
when shutter is
closed)
CMN Imaging Core
January 16, 2013
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LCD display panel
Indicates
position of
objective
relative to
upper limit
Indicates
current
step size
setting for
focusing
arrow
indicates
upper limit
has been
set
arrow
indicates
lower limit
has been
set
Indicates
current filter
cube
Indicates
current
objective
Indicates
desired DIC
objective
prism
D= dry mode
I= immersion mode
Indicates
actual DIC
objective
prism in light
path
CMN Imaging Core
January 16, 2013
9
LCD display panel buttons
Do not press the LEARN button (this will enter you
into learn mode, which you must not play with). If you
accidentally press this button, you will see the word
“EXIT “ flashing. Press the LEARN button again to
exit the learn mode.
Learn mode
(DO NOT
USE!)
Set lower
limit of
travel for
objective
(DO NOT
USE!)
Toggle
between
voltage
readout
and
objective
readout
Adjust step
size for
coarseness
of focusing
Set upper
limit of
travel for
objective
(DO NOT
USE!)
CMN Imaging Core
January 16, 2013
10
Changing filters
Use the motorized fluorescent filter cube changer on
the microscope control panel:
Red light shows current filter cube selection
The filter cube turret contains three filters plus an
empty slot
GFP : cube for green fluorochromes
RED : cube for red fluorochromes
CFP/ : cube for far red fluorochromes
or cube for CFP (see Paula for details)
SCAN : empty slot
Notes:
The cube in the CFP/ position is normally the far
red cube, suitable for fluorochromes such as
TOTO-3 and Cy5
The SCAN position is used for confocal imaging
and for transmitted light observation through the
eyepieces
Press left button
to rotate filter
cube turret left
Press right button
to rotate filter
cube turret right
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