Leica DMI4000B User manual

Leica DMI3000B,
DMI4000B, DMI6000B
Instructions · Bedienungsanleitung · Mode d’emploi

Published August 2010 by:
Herausgegeben August 2010 von:
Edité en août 2010 par :
Leica Microsystems CMS GmbH
Ernst-Leitz-Straße 17-37
D-35578 Wetzlar (Germany)
Responsible for contents:
Verantwortlich für den Inhalt:
Responsable du contenu rédactionnel :
Bernard Kleine
(Marketing CMS, Life Science Research Microscopy, Product
Management)
(Marketing CMS, Life Science Research Microscopy, Produkt-
management)
(Marketing CMS, Life Science Research Microscopy,
chef de produit)
Dietmar Gnass
(R&D Manager)
In case of questions, please contact the hotline:
Bei Fragen wenden Sie sich bitte an die Hotline:
Pour toute question, contacter notre service d’assistance
téléphonique : Phone: [phone] - 29 42 53
Fax: [phone] - 29 22 55
E-Mail: [email protected]

3
Leica DMI3000B,
DMI4000B, DMI6000B
Instructions

4
Copyrights
Copyrights
All rights to this documentation are held by Lei-
ca Microsystems CMS GmbH. Reproduction of
text or illustrations (in whole or in part) by print,
photocopy, microfilm or other method (including
electronic systems) is not allowed without ex-
press written permission from Leica Microsys-
tems CMS GmbH.
The term „Windows“ may appear in the following
text without further identification. It is, however,
a registered trademark of Microsoft Corpora-
tion. The names of companies and products used
herein may be trademarks of their respective
owners.
The instructions contained in the following docu-
mentation reflect state-of-the-art technology
standards. We have compiled the texts and illus-
trations as accurately as possible. Nevertheless,
no liability of any kind may be assumed for the
accuracy of this manual’s contents. Still, we are
always grateful for comments and suggestions
regarding potential mistakes within this docu-
mentation.
The information in this manual is subject to modi-
fication at any time and without notification.

5
Contents
6.10 Installation and Replacement of the
transmitted Light Lamps:
107 or 107/2 Lamp Housing........................ 45
6.11 Installing the Lamp Housing Mount
and Mirror Housing)................................... 46
6.12 Installation and Replacement of
Incident Light Lamps.................................. 48
6.13 Equipping the Incident Light
Turret Disk.................................................... 52
6.14 Inserting the Front Module Slider............ 55
6.15 Installation of the Polarizer
and Analyzer................................................ 55
6.16 Optional Accessories................................. 57
6.17 Connection to the Electronics Box.......... 58
6.18 Connection to the Computer..................... 59
6.19 Connection to the Power Supply ............. 59
7. Start-up ........................................................ 60
7.1 Functional Principle .................................. 60
7.2 Switching on the Microscope.................. 64
7.3 The LeicaDisplay ........................................ 65
7.4 The Function Buttons on the Stand......... 66
7.5 The SmartMove
Remote Control Module ............................ 69
7.6 Illumination.................................................. 69
7.6.1 Transmitted light .............................. 69
7.6.2 Incident Light - Fluorescence........ 73
7.7 Checking Phase Contrast Rings............... 74
7.8 Checking modulation contrast
slit diaphragms............................................ 77
7.9 Setting the Motorized Polarizer ............... 77
7.10 Adjusting the Light Sources ..................... 78
Contents
1. Important Notes about this Manual........ 7
2. Intended Purpose of the Microscope..... 8
3. Safety Notes................................................ 9
3.1 General Safety Notes ................................ 9
3.2 Electrical Safety.......................................... 10
3.3 Safety Instructions
for Handling the Light Sources ................ 12
3.4 Notes on handling laser devices ............. 12
3.5 Safety Instructions
for Handling Acids and Bases.................. 12
3.6 Disposal........................................................ 13
4. Overview of the Leica DMI Series.......... 14
5. Unpacking the Microscope ..................... 27
6. Assembling the Microscope.................... 30
6.1 Assembly Tools........................................... 30
6.2 Installation of the Transmitted Light
Illumination Carrier (TL)............................. 31
6.3 Installation of the DIC Module
and DIC Objective Prisms ......................... 32
6.4 Installation of Specimen Stages.............. 33
6.5 Installation of Condensers........................ 38
6.6 Installation of Eyepieces........................... 43
6.7 Installation of Objectives .......................... 43
6.8 Installation of Filters
in the Illumination Arm............................... 44
6.9 Installing the transmitted Light
Lamp Housing ............................................. 44

6
Contents
8. Operation..................................................... 81
8.1 Switching on................................................ 81
8.2 Contrast Methods....................................... 83
8.2.1 Bright Field (TL)................................ 83
8.2.2 Phase Contrast (TL) ....................... 85
8.2.3 Dark Field (TL) .................................. 86
8.2.4 Polarization (TL)............................... 87
8.2.5 Differential
Interference Contrast (TL) ............. 88
8.2.6 Integrated Phase Contrast (TL)..... 89
8.2.7 Integrated
Modulation Contrast (TL)................ 90
8.3 Fluorescence............................................... 91
8.4 Combination Methods ............................... 93
8.5 Focusing....................................................... 94
8.6 Tubes ............................................................ 96
8.7 Port selection ............................................. 96
8.8 Eyepieces..................................................... 97
8.9 Objectives.................................................... 98
8.10 Stages and Object Displacement ............ 101
8.11 Magnification Changer.............................. 102
8.12 Light sources............................................... 103
8.13 Aperture and Field Diaphragm ................ 104
9. Troubleshooting.......................................... 105
10. Care of the Microscope ............................ 109
10.1 Dust Cover ................................................... 109
10.2 Cleaning ....................................................... 109
10.3 Handling Acids and Bases........................ 110
11. Major Consumable
and Replacement Parts............................. 111
12. Dimensions.................................................. 112
13. Abbreviations and Pictograms................ 113
14. Index ............................................................ 115
15. EU Declaration of Conformity .................. 117

7
1. Important Notes about this Manual
(1.2)
→p. 20
!
*
Numbers in parentheses, such as „(1.2)“, corre-
spond to illustrations (in the example, Figure 1,
Item 2).
Numbers with pointer arrows (for example
→ p. 20), point to a certain page of this manual.
Caution!
Special safety instructions within this manual
are indicated with the triangle symbol shown
here, and have a gray background.
Caution! The microscope and accessories can
be damaged when operated incorrectly.
Notes on the disposal of the microscope, ac-
cessories and consumable materials.
Explanatory note.
Item not contained in all configurations.
Text symbols, pictograms and their meanings:
Caution!
This operating manual is an essential com-
ponent of the microscope, and must be read
carefully before the microscope is assem-
bled, put into operation or used.
1. Important Notes about this Manual
This operating manual contains important in-
structions and information for the operational
safety and maintenance of the microscope and
accessories. It must therefore be kept safely for
future reference.
A separate manual is available on CD-ROM cov-
ering the operation of the Leica Application Suite
(LAS).

8
2. Intended Purpose of the Microscope
2. Intended Purpose of the Microscope
The Leica DMI Series microscopes covered in
this manual are designed for biological, routine,
and research applications. This includes the ex-
amination of samples taken from the human body
in order to provide information on physiological
or pathological states or congenital abnormali-
ties; to determine the safety and compatibility
with potential recipients; or to monitor therapeu-
tic measures.
The Leica DMI Series is an additional develop-
ment of Leica’s proven inverted research mi-
croscopes, designed for cellular and tissue
examination, micromanipulation and microinjec-
tion techniques, microdissection, and confocal
microscopy. The Leica DMI Series is suitable
for universal deployment. All contrast methods
such as dark field, bright field, phase contrast,
DIC, fluorescence, and modulation contrast are
integral to the microscope and can be adapted
or changed quickly and easily. Variable illumi-
nation and imaging beam paths, as well as HCS
optics, modular accessories, and a comprehen-
sive range of peripherals complement the Leica
Microsystems inverted research stand.
The above-named microscope series complies
with the Council Directive 98/79/EEC concern-
ing in vitro diagnostics. They also conform to the
Council Directives 73/23/EEC concerning electri-
cal apparatus and 89/336/EEC concerning elec-
tromagnetic compatibility for use in an industrial
environment.
Caution!
The manufacturer assumes no liability for
damage caused by, or any risks arising from,
using the microscopes for purposes other
than those for which they are intended or not
using them within the specifications of Leica
Microsystems CMS GmbH.
In such cases the declaration of conformity
shall cease to be valid.
Caution!
These (IVD) devices are not intended for use
in the patient environment defined by DIN
VDE 0100-710. Neither are they intended for
combining with medical instruments accord-
ing to EN 60601-1. If a microscope is electri-
cally connected to a medical instrument
according to EN 60601-1, the requirements
defined in EN 60601-1-1 shall apply.
8

9
3. Safety Notes
3. Safety Notes
3.1 General Safety Notes
This safety class 1 device is constructed and
tested in accordance with
EN 61010-2-101:2002,
EN 61010-1:2001,
IEC 61010-1:2001,
Safety regulations for electrical measuring, con-
trol, and laboratory devices.
Caution!
In order to maintain this condition and to en-
sure safe operation, the user must follow the
instructions and warnings contained in this
operating manual.
Caution!
The devices and accessories described in
this operating manual have been tested for
safety and potential hazards.
The responsible Leica affiliate or the main
plant in Wetzlar must be consulted whenever
the device is altered, modified or used in con-
junction with non-Leica components that are
outside of the scope of this manual.
Unauthorized alterations to the device or
noncompliant use shall void all rights to any
warranty claims!
9

10
3. Safety Notes
3.2 Electrical Safety
General Specifications
Leica CTR4000, CTR5000, CTR5500, CTR6000,
CTR6500, CTR7000, CTR6500 HS, CTR7000 HS
Electronics Boxes
For indoor use only.
Supply voltage:
Frequency:
Power input:
Fuses:
Ambient temperature:
Relative humidity:
Over voltage category:
Pollution degree:
Microscope
For indoor use only.
Supply voltage:
Frequency:
Power input:
Fuses:
Ambient temperature:
Relative humidity:
Over voltage category:
Pollution degree:
90–250 V~
50–60 Hz
max. 290 VA
T6.3 A
(IEC 60127-2/3)
15–35°C
max. 80% to 30°C
II
2
90–250 V~
50–60 Hz
see CTR4000–7000
see CTR4000–7000
15–35°C
max. 80% to 30°C
II
2
ebq 100 supply unit*
For indoor use only.
Supply voltage:
Frequency:
Power input:
Fuses:
Ambient temperature:
Relative humidity:
Over voltage category:
Pollution degree:
(see enclosed manual)
Leica EL6000*
For indoor use only.
Supply voltage:
Frequency:
Power input:
Fuses:
Ambient temperature:
Relative humidity:
Overvoltage category:
Pollution degree:
(see enclosed manual)
100–240 VAC
50–60 Hz
max. 200 VA
5x20, 2.5 A, slow,
breaking capacity H
0°–40°C
10–90%
non-condensing
II
2
90–250 V~
50–60 Hz
See CTR4000–7000 HS
See CTR4000–7000 HS
15–35°C
max. 80% to 30°C
II
2

11
3. Safety Notes
Caution!
The microscope’s electrical accessory com-
ponents are not protected against water. Wa-
ter can cause electric shock.
Caution!
Protect the microscope from excessive tem-
perature fluctuations. Such fluctuations can
lead to the accumulation of condensation,
which can damage the electrical and optical
components.
Ambient temperature: 15–35°C.
Caution!
Before exchanging the fuses or lamps, be ab-
solutely certain to switch off the main power
switch and remove the power cable.
Caution!
Power plugs may only be plugged into an out-
let equipped with a grounding contact.
Do not interfere with the grounding function
by using an extension cord without a ground
wire. Any interruption of the ground wire in-
side or outside of the device, or release of
the ground wire connection, can cause the
device to become hazardous. Intentional
ground interruption is not permitted!
Caution!
Peripheral devices with their own or sepa-
rate power supplies that are connected to
the microscope can have the same protec-
tive conductor potential by connecting them
to the ground screw on the back of the Leica
CTR4000, CTR6000, CTR6500 and CTR7000
electronics boxes. For connections without
a ground connector, Leica Service must be
consulted.
Caution!
Never use any fuses as replacements other
than those of the types and the current rat-
ings listed here. Using patched fuses or
bridging the fuse holder is not permitted. The
use of incorrect fuses may result in a fire haz-
ard.

12
3. Safety Notes
3.3 Safety Instructions
for Handling the Light Sources
Caution!
Light sources pose a potential irradiation risk
(glare, UV-radiation, IR-radiation). Therefore,
lamps have to be operated in closed hous-
ings.
Never look directly into the beam path (blind-
ing hazard).
Connect the light guide to the microscope
first to prevent exposing the user to the high-
energy light output of the Leica EL6000 com-
pact light source.
Never look directly into the light emitted by
the light guide.
3.4 Notes on handling laser devices
The microscope is not suitable for coupling laser
devices into the camera ports (refer to Chapter
4), as this creates a danger to the user from laser
radiation.
For use of the microscope with lasers, Leica
Microsystems offers special microscopes with
additional safety devices.
For further information, please contact your au-
thorized Leica Microsystems representative.
Caution!
Follow safety instructions for immersion oil!
3.5 Safety Instructions
for Handling Acids and Bases
For examinations using acids or other aggressive
chemicals, particular caution must be taken.
Caution!
Be absolutely certain to prevent coming into
contact with these chemicals.

13
3. Safety Notes
3.6 Disposal
To dispose of the product at the end of its service
life, please contact Leica Service or Sales.
Please observe national laws and regulations,
such as those implementing and enforcing the
WEEE EU Directive.
Note!
Like other electronic devices, the micro-
scope, its accessories and consumable ma-
terials must not be disposed of as regular
household waste.

14
4. Overview of the Instruments
4.1 Specifications
4. Overview of the Leica DMI Series
Contrast Methods
Transmitted Light Axis
Leica DMI Series
• transmitted light (TL): BF, DF, PH, DIC, Pol
• intermediate pupil: IMC (integrated modulation contrast)
IPH (Integrated phase contrast)
• incident light (IL): Fluo
Leica DMI4000 B and DMI6000 B
• combination (TL/IL): Fluo/DIC, Fluo/PH
Leica DMI Series
• Manual and coded transmitted light illumination arm with inte-
grated mechanical tilt mechanism to provide adequate space for
specimens and micromanipulators, integrated field diaphragm,
filter magazine for 2 replaceable filters, condenser quick-chang-
er
•
Illumination Manager (aperture diaphragm, field diaphragm, light in-
tensity)
• manual shutter
• lamp housing mount for interchangeable lamp housings.
• with integrated cable channel
Leica DMI4000 B and Leica DMI6000 B
• Motorized or manual/coded transmitted light illumination arm
with integrated mechanical tilt mechanism to provide adequate
space for specimens and micromanipulators, integrated motor-
ized field diaphragm, motorized filter magazine for 2 replaceable
filters, condenser quick-changer
• with integrated cable channel
• automatic Illumination Manager
(aperture, field diaphragm, intensity, process switching)
• manual or motorized shutter
• lamp housing mount for interchangeable lamp housings.
• automatic, electronic condenser identification

15
4. Overview of the Instruments
Leica DMI3000 B
• manual shutter
• lamp housing mount for up to 3 interchangeable light sources
• manual 5-place filter turret
• Fluorescence Intensity Manager (FIM)
(reduction of incident illumination intensity)
Leica DMI4000 B and Leica DMI6000 B
• automatic Illumination Manager
(aperture, field diaphragm*, intensity, process switching)
• motorized shutter (switching speed < 50 ms)
• lamp housing mount for up to 3 interchangeable light sources
• motorized 6-place filter turret
• Fluorescence Intensity Manager (FIM)
(reduction of incident illumination intensity)
• Optional: Interface for structured illumination
• Leica DMI6000 B:
mechanical booster lens for central boosting of
fluorescence or uniform distribution
• motorized Excitation Manager* to monitor fluorescence emission
when using double and triple filter cubes
• ultra fast filter wheel for 3 excitation wavelengths
(switching speed < 50 ms)
Leica DMI Series
• ergonomic with or without camera port at left
• 2 switching positions: 100%VIS and 50%VIS / 50%CAM or
• 2 switching positions: 100%VIS and 0%VIS / 100%CAM
• optional Bertrand lens
• eye spacing adjustment
• height and angle adjustment (30° - 45°)
Leica DMI4000 B and Leica DMI6000 B
• motorized
• 3 switching positions
(choice of magnifications: 1x; 1.5x; 1.6x or 2.0x)
• effective on all camera ports and eyepieces
or Leica DMI Series
• manual
• 2 switching positions
(choice of magnifications: 1x; 1.5x; 1.6x or 2.0x)
• effective on tube port and eyepieces
Incident Light Axis
Tube
Magnification Changer
* not in combination with structured Illumination

16
4. Overview of the Instruments
Leica DMI6000 B
• motorized and coded
• 6x for objectives with M25 thread and 45 mm parfocal distance
• for DIC: motorized or manual/coded Wollaston prism carousel
• anti-vibration locking
Leica DMI4000 B
• manual and coded
• 6x for objectives with M25 thread and 45 mm parfocal distance
• for DIC: motorized or manual/coded Wollaston prism carousel
Leica DMI3000 B
• manual
• 6x for objectives with M25 thread and 45 mm parfocal distance
• for DIC: manual Wollaston prism carousel
Leica DMI Series
Fixed regular stages
• Ceramic-coated stage plate (248 mm x 204 mm)
• heating stage plate (3°C above room temperature to 60°C)
(248 x 212 mm)
• temperature-controlled stage plate (0°C to 60°C)
(248 mm x 212 mm)
• fixed micromanipulation stages
• ceramic-coated stage plate (248 mm x 204/122 mm)
• heated stage plate (from 3°C above room temperature
to 60°C) (248 mm x 204/122 mm)
• temperature-controlled stage plate (0°C to 60°C)
(248 mm x 204/122 mm)
• regular manual and motorized 3-plate cross-stage
• positioning range: 83 mm x 127 mm
• 20 optional inserts (standard, heating, cooling) for a variety
of applications, size of inserts:160 mm x 110 mm
(compatible with scanning stages)
• narrow manual and motorized micromanipulation
3-plate cross-stage
• positioning range: 40 mm x 40 mm
• 3 optional inserts for a variety of applications
• Scanning stage 120 x 100 (motors on bottom)
• 1 mm, 2 mm, 4 mm spindle pitch
(higher resolution vs. higher speed)
• 20 optional inserts (standard, heating, cooling) for a variety
of applications, size of inserts:160 mm x 110 mm
Objective Turret
Stages

17
4. Overview of the Instruments
Condensers
Z Focus
Observation Ports
Leica DMI4000 B and Leica DMI6000 B
(identical for Leica DMI3000 B, but manual)
• motorized and coded or manual and coded
• motorized or manual aperture diaphragm
• contrast methods: BF, DF, PH, DIC, Pol, IMC, IPH
• automatic method switching
• condenser turret with 7 positions for contrast methods
• 2 condenser housings (S1-S28 and S40,S70)
• condenser heads: S1/1.4 oil, S1/0.9 dry, S23/0.53, S28/0.55
• condenser heads can be swung out
• condenser S40/S70 with additional lens for low magnifications
• all condensers suitable for magnifications from 1.25x to 100x
• with or without motorized or manual polarizer
• with or without motorized or coded Wollaston prism disk
Leica DMI6000 B
• motorized and coded
• 9 mm travel (1 mm below, 8 mm above the stage)
• maximum travel speed: 5 mm/s
• 5 focus steps: 0.05 µm; 0.1 µm; 0.7 µm; 1.5 µm; 5.0 µm
• electronic focus repositioning
• automatic lowering prior to objective change
• electronic parfocality
• Optional: Adaptive Focus Control (AFC)
Leica DMI3000 B and Leica DMI4000 B
• manual
• 9 mm travel (1 mm below, 8 mm above the stage)
Leica DMI6000 B
• motorized and coded
• left side ports (100%, 80% or 50% transmission)
• left side port dichroic splitting at 680 nm
• right side ports (100%, 80% or 50% transmission)
• bottom port
optional
• top port with 2 switching positions
• 100% to eyepieces
• 50% to eyepieces/ 50% to port
Leica DMI4000 B
left side port, manual (100% or 80% transmission)

18
4. Overview of the Instruments
Observation Ports
Controls
Electronics Box
Leica DMI3000 B
(a manual side port is a standard feature of the Leica DMI3000 B stand)
• manual
• left side port (80% or 100% transmission)
Leica DMI4000 B and Leica DMI6000 B
• 7 fixed control buttons for illumination and apertures
• 7 variable function buttons behind the focus controls
• 3 fixed control buttons for focus stops (Leica DMI6000 B only)
• 2 focus hand wheels
• 7 buttons for fluorescence cubes and shutters
• 4 buttons for magnification changer and ports
• SmartMove: ergonomic remote control module for x,y,z control
and four additional variable function buttons
• STP6000
Leica DMI3000 B
• 2 focus hand wheels
• 1 illumination hand wheel
• 2 turning knobs for field diaphragm and FIM adjustment
• 1 On/Off switch
• separate control unit for all motorized and electronic elements of
the microscope such as:
For CTR6500 (HS)/CTR7000 (HS) only
• scanning stages
For CTR6000 only
• motorized 3-plate cross-stages
For CTR6000/7000
• objective turret
• focus
• ports
• magnification changer
• fluorescence
• condenser
• power supply for SmartMove
For all CTR boxes
with
• power supply for 100W halogen lamps

19
4. Overview of the Instruments
Leica DMI4000 B and Leica DMI6000 B
• 2 x RS232C
• 2 x USB
• 4 x external/internal peripherals
• CTR boxes
• SmartMove
• STP6000
Leica DMI4000 B and Leica DMI6000 B
• Leica Application Suite (LAS) for WindowsTM
with plug-ins for:
• microscope and camera configuration
• microscope and camera control
• image acquisition
Interfaces
Software Tools

20
4. Overview of the Instruments
Fig. 1 Left side, Leica DMI4000 B and DMI6000 B
1Eyepiece
2Eyepiece tube
3Top port
4Intermediate pupil interface
5 LeicaScreen
6Light intensity
7Field diaphragm
8TL/IL switching
9Aperture diaphragm
10 Focus wheel (motorized Leica DMI6000 B,
manual (fine and coarse) Leica DMI4000 B)
11 Variable function buttons
12 Left side port
13 Booster lens
(Leica DMI6000 B fluorescence microscopes only)
14 Lamp mount (fluorescence microscopes only)
15 Condenser head
16 Condenser base
17 Field diaphragm
18 Transmitted light lamp housing
19 DIC objective prism disk
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