Leica DMIRE2 Manual de usuario

CMN Imaging Core

January 16, 2013

LEICA DM IRE2 MICROSCOPE MANUAL

Neuroscience Imaging Core

Rightmire Hall

Ohio State University

Director: Tony Brown

Rightmire 060

292-1205

[email protected]

Facility Manager: Paula Monsma

Rightmire 062

292-3025

[email protected]

This manual prepared by Tony Brown.

CMN Imaging Core

January 16, 2013

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

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

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

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

OBJECTIVE PRISM TURRET POSITIONS

Objective

turret

position

Description

Matching objectives

Empty (for bright field

or phase contrast)

DIC objective prism

20x multi-immersion

DIC objective prism

63x water

immersion

100x oil immersion

DIC objective prism

40x oil immersion

63x oil immersion

CONDENSER TURRET POSITIONS

Condenser

turret

position

Description

Matching objectives

Empty (for bright

field)

PH1

phase ring

10x dry

PH2

phase ring

40x dry

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

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

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

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

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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