Leica DMIRE2 Manual de usuario

Cell Imaging Unit – UIC

LEICA DMIRE2 MICROSCOPE MANUAL

TABLE OF CONTENTS

ANATOMY OF THE LEICA DMIRE2 INVERTED MICROSCOPE 3

INITIALISATION PROCEDURE 4

LEFT SIDE OF MICROSCOPE (DETAILED VIEW) 5

DMIRE2 FRONT PANEL 6

LCD DISPLAY OF MICROSCOPE STATUS 6

LCD DISPLAY CONTROL BUTTONS 7

L.E.D. PANEL SHOWING ACTIVE FLUORESCENCE CUBE 7

FILTER CUBE SELECTION SWITCH 7

MERCURY LAMP SHUTTER 8

PORT SELECTION SWITCH 8

TUBE LENS 8

CONTRAST METHODS 9

BRIGHTFIELD 9

FLUORESCENCE 9

MAGNIFICATION 11

CHANGING OBJECTIVE 11

FOCUSSING 12

SCANNING MODE 13

ANATOMY OF THE LEICA DMIRE2 INVERTED MICROSCOPE

1. Binocular phototube 16. Filters

2. Eyepiece adapter tube 17. Aperture diaphragm adjustment

3. Eyepieces 18. Lamp housing mount

4. Tube mount 19. Lamp housing

5. Tube port for photo/TV connect 20. Stage plate

6. Beam splitter 21. Analyser

7. Mains switch 22. Tube lens module

8. Brightness adjustment 23. Switch rod for lateral TV port

9. Lateral TV port 24. Transmitted light illumination

10. Coaxial coarse & fine focus 25. Condenser

11. Fluorescence module 26. Transmitted light lamp housing

12. ICT prism adjustment 27. Transmitted light field diaphragm

13. Sextuple objective nosepiece 28. SLR port

14. Centring buttons for incident light 29. Second lamp housing

15. Field diaphragm adjustment

INITIALISATION PROCEDURE

1. Ensure that both the DMIRE2 control box (CTRMIC) and the mercury arc lamp

power supply have been switched on (shown below). After switching on the

CTR6000 control box the microscope will take a few seconds to initialize

2. During microscope initialisation the stage position will be reset and recently used

settings will be recalled. After initialisation the microscope status menu will appear

on the L.C.D. display screen (shown below).

LEFT SIDE OF MICROSCOPE (DETAILED VIEW)

A Halogen lamp intensity adjustment wheel

B Coaxial coarse & fine focus

*Beam splitter (not functional)

C Epifluorescence illumination diaphragm (controls brightness of illumination)

DEpifluorescence field diaphragm (controls area of illumination)

EObjective prism turret

FFilter cube turret

DMIRE2 FRONT PANEL

1. LCD DISPLAY OF MICROSCOPE STATUS (ALPHA-NUMERIC).

Displays the following information:

•Position of objective lens relative to upper limit (e.g. -145µm)

•Step size setting for focussing (S0-3)

•Upper & lower limits for stage position (↑↓)

•Current objective settings (5x, 10x, 25x, 40x, 63x)

•Current filter cube. For brightfield (BF) or fluorescence optics (A, N21, I3; below)

•Dry/ immersion mode (D/I)

2. LCD DISPLAY CONTROL BUTTONS

LEARN Puts microscope in learn mode (DO NOT USE). If you press

this button by accident, you will see the word “EXIT” flashing.

Press the LEARN button again to exit the learn mode.

CHANGE Toggles between voltage readout and objective readout.

Set lower limit of travel for objective (DO NOT USE)

Set upper limit of travel for objective (DO NOT USE)

STEP Adjust step size for coarseness of focussing. Four settings are

available (S0, fine; S1, medium fine; S2, medium coarse & S3,

coarse).

3. L.E.D. PANEL SHOWING ACTIVE FLUORESCENCE FILTER CUBE

FILTER COLOUR OF LIGHT FLUOROCHROMES

(1) --------------- ---------------

(2)

DAPI 31000UZ

UV light (BP350/50) DAPI

(3)

GFP

Blue light (BP470/40) FITC, Alexa 488

(4)

mRFP

Green light (BP 584/50) TRITC, Texas Red, Alexa 546

4. FILTER CUBE SELECTION SWITCH (TWO-WAY)

Pressing the left button will rotate the motorised filter cube turret left, whereas the

right switch will rotate the filter cube turret right. The active filter cube will be

illuminated (red) in the horizontal LED panel (see above).

5. MERCURY LAMP SHUTTER SWITCH (ONE WAY)

Red LED is on when shutter is closed

6. PORT SELECTION SWITCH (TWO WAY)

VIS Light directed to binocular eyepieces

SIDE Light directed to scanner

BOTTOM Disabled

The active port will be illuminated (red) in the vertical LED panel.

CONTRAST METHODS

BRIGHTFIELD

Use the wheel on the front left side of the microscope stand near the base to turn on

and adjust the halogen lamp brightness (shown below).

The lamp voltage will be displayed automatically on the microscope control panel

when the intensity is adjusted. To switch off transmitted light illumination adjust lamp

intensity to 2.5V then continue rotating the wheel beyond this point. 0V on control

panel display indicates illumination is off. To switch on, rotate the wheel in the

opposite direction.

FLUORESCENCE

Ensure transmitted light has been switched off (0V on LCD display panel) and the

mercury lamp is switched on. Cycle through fluorescent cubes by pressing the

fluorescent cube selection switches (*).

Press the shutter switch to close the mercury lamp shutter (red light is on when shutter

is closed).

MAGNIFICATION

The following objective lenses are fitted on the DMIRE2:

TUR

Ret

pos

Objective type Mag N.A. Immersion Cover-glass

DIC prisms

(upper S1)

DIC

prisms

(upper

S23)

DIC

prisms

(lower)

Pixel size

(1xc-

mount)

(um/pixl)

1 N PLAN 2,5x 0.07 DRY With/without -K1 C 2.58

2 HC PL FLUOTAR 10x 0.30 DRY With /without Not avail. (K11) K3 A 0.64

3 HC PL APO 20x 0.70 DRY 0.17 #1.5 K3 K3 D

4 HCX PL APO 40x 0.85 DRY 0,17 #1.5 K3 K7 A 0.16

5 HCX PL APO CS 63x 1.30 GLYC 0.17 #1.5 K7 K7 D

6 HCX PL APO 100x 1.40-

0.70 OIL 0,17 # 1.5 K7 Not avail.

(K10)

HCX APO

40x 0.80 WATER With /without - - - -

CHANGING OBJECTIVE

To reduce the chance of immersion medium getting on to a dry objective, the

microscope will not allow you to move freely between oil immersion and dry lens

mode. To switch from one mode to another, simultaneously press the upper limit and

lower limit buttons on the microscope control panel (asterisks).

The words “CHANGE OBJECTIVE” will flash on the LCD display. Now you can

change the objective using the objective turret control buttons located behind the

coaxial coarse and fine focus controls on the LEFT side of the microscope (upper

button increases the magnification; lower button decreases the magnification)

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