Leica DMI6000 B User manual

1
Leica DMI6000 B
Operating Manual

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Published October 2004 by:
Leica Microsystems Wetzlar GmbH
Ernst-Leitz-Straße
D-35578 Wetzlar (Germany)
Responsible for contents:
Bernard Kleine
(Marketing CM, Compound Microscopy, Product Manage-
ment)
Holger Grasse
(Safety Officer according to MPG §30) In case of questions, please contact the hotline:
Phone [phone]
Fax [phone]
E-mail: [email protected]

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Leica DMI6000 B
Operating Manual

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Copyrights
Copyrights
All rights to this documentation are held by
Leica Microsystems Wetzlar GmbH. Reproduc-
tion of text or illustrations (in whole or in part) by
print, photocopy, microfilm or other method (in-
cluding electronic systems) is not allowed with-
out express written permission from Leica Mi-
crosystems Wetzlar 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 respec-
tive owners.
The instructions contained in the following doc-
umentation reflect state-of-the-art technology
and knowledge standards. We have compiled
the texts and illustrations as accurately as pos-
sible. Nevertheless, no liability of any kind may
be assumed for the accuracy of this manual’s
contents. Still, we are always grateful for com-
ments and suggestions regarding potential mis-
takes within this documentation.
The information in this manual is subject to modifi-
cation at any time and without notification.

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Contents
6.11 Installation of Lamp Housing Mount
and Mirror Housing ................................... 42
6.12 Installation and Replacement
of Incident Light Lamps ............................ 44
6.13 Equipping the Incident Light
Turret Disk ................................................... 48
6.14 Installation of the Polarizer and
Analyzer....................................................... 50
6.15 Optional Accessories ............................... 52
6.16 Connection to the
Electronics Box CTR6000 ......................... 53
6.17 Connection to the Computer ................... 54
6.18 Connection to the Power Supply ............ 54
7. Start-up........................................................ 55
7.1 Functional Principle .................................. 55
7.2 Switching on the Microscope................. 59
7.3 The LeicaScreen........................................ 60
7.4 The Function Buttons on the Stand ....... 61
7.5 The SmartMove Remote Control
Module......................................................... 64
7.6 Illumination ................................................. 65
7.6.1 Transmitted Light ............................ 65
7.6.2 Incident Light - Fluorescence ...... 68
7.7 Checking Phase Contrast Rings ............. 69
7.8 Setting the Motorized Polarizer............. 70
7.9 Adjusting the Light Sources .................... 71
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
4. Overview of the Leica DMI
6000 B......... 12
4.1 Specifications ............................................ 12
4.2 Glossary....................................................... 16
5. Unpacking the Microscope .................... 22
6. Assembling the Microscope .................. 25
6.1 Assembly Tools .......................................... 25
6.2 Installation of the Transmitted Light
Illumination Carrier (DL) ........................... 26
6.3 Installation of the DIC Module
and DIC Objective Prisms ........................ 27
6.4 Installation of Specimen Stages............. 28
6.5 Installation of Condensers....................... 34
6.6 Installation of Eyepieces.......................... 39
6.7 Installation of Objectives ......................... 39
6.8 Installation of Filters
in the Illumination Arm ............................. 40
6.9 Installation of the
Transmitted Light Lamp Housing ............ 40
6.10 Installation and Replacement of the
Transmitted Light Lamps: .........................
Lamp Housing 107 or 107/2 ...................... 41

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Contents
8. Operation .................................................... 74
8.1 Switching On .............................................. 74
8.2. Contrast Methods...................................... 76
8.2.1 Brightfield (TL)................................. 76
8.2.2 Phase Contrast (TL) ........................ 77
8.2.3 Darkfield (TL) ................................... 77
8.2.4 Polarization (TL) .............................. 78
8.2.5 Differential
Interference Contrast (TL) ............ 79
8.3 Fluorescence.............................................. 80
8.4 Combination Methods .............................. 81
8.5 Focusing ...................................................... 82
8.6 Tubes............................................................ 84
8.7 Eyepieces .................................................... 85
8.8 Objectives ................................................... 85
8.9 Stages and Object Displacement ........... 88
8.10 Magnification Changer............................. 89
8.11 Light Sources ............................................. 89
8.12 Aperture and
Field Diaphragm ......................................... 90
9. Troubleshooting......................................... 91
10. Care of the Microscope ........................... 95
10.1 Dust Cover .................................................. 95
10.2 Cleaning....................................................... 95
10.3 Handling Acids and Bases ...................... 96
11. Major Consumable and Replacement
Parts ............................................................ 97
12. Dimensions................................................. 98
13. Abbreviations and Pictograms............... 99
14. Index ............................................................ 101
15. EU Declaration of Conformity ................. 104

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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 manu-
al are indicated with the triangle symbol
shown here, and have a gray background.
Caution! The microscope and accessories can
be damaged when operated incorrectly.
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 6000 B microscope described in
these instructions is intended for biological rou-
tine and research applications. This includes the
examination of samples taken from the human
body to provide information on physiological or
pathological states or congenital abnormalities,
or determing the safety and compatibility with po-
tential recipients, or monitoring therapeutic
measures.
The Leica DMI
6000 B is a further development
of Leica’s proven inverted research micro-
scopes. It is designed for cellular and tissue ex-
amination, micromanipulation and microinjec-
tion techniques, microdissection, and confocal
microscopy. The Leica DMI
6000 B is suitable for
universal deployment, all contrast methods such
as darkfield, brightfield, phase contrast, DIC, flu-
orescence, and modulation contrast are integral
to the microscope and can be adapted or
changed quickly and easily. Variable illumination
and imaging beam paths, as well as HCS optics,
modular accessories and a comprehensive
range of peripherals, complement the Leica DMI
6000 B inverted research stand.
The above-named microscope complies with
the Council Directive 98/79/EEC concerning in
vitro diagnostics. It also conforms to the Council
Directives 73/23/EEC concerning electrical ap-
paratus and 89/336/EEC concerning electromag-
netic compatibility for use in an industrial envi-
ronment.
Caution!
The manufacturer assumes no liability for
damage caused by, or any risks arising from
using the microscope for other purposes
than those for which it is intended or not us-
ing it within the specifications of Leica Mi-
crosystems Wetzlar GmbH.
In such cases the declaration of conformity
shall cease to be valid.
Caution!
This (IVD) device is not intended for use in
the patient environment defined by DIN VDE
0100-710. Neither is it intended for combin-
ing with medical instruments according to
EN 60601-1. If a microscope is electrically
connected to a medical instrument accord-
ing to EN 60601-1, the requirements de-
fined in EN 60601-1-1 shall apply.

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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 1010-1:2001,
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 whenev-
er the device is altered, modified, or used in
conjunction 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!

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3. Safety Notes
Caution!
Power plugs may only be plugged into an
outlet 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
CTR6000 electronics box. 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
hazard.
90-250V~
50-60 Hz
max. 290VA
T6.3 A
(IEC 60127-2/3)
15-35°C
max. 80% to 30°C
II
2
90-250V~
50-60 Hz
See CTR6000
See CTR6000
15-35°C
max. 80% to 30°C
II
2
90-250V~
50-60 Hz
max. 155VA
2xT2A (IEC 127)
10-36°C
max. 80% to 30°C
II
2
3.2 Electrical Safety
General specifications
Leica CTR6000 Electronics Box
For indoor use only.
Supply voltage:
Frequency:
Power input:
Fuses:
Ambient temperature:
Relative humidity:
Overvoltage category:
Pollution degree:
Microscope
For indoor use only.
Supply voltage:
Frequency:
Power input:
Fuses:
Ambient temperature:
Relative humidity:
Overvoltage category:
Pollution degree:
ebq 100 Supply Unit*
For indoor use only.
Supply voltage:
Frequency:
Power input:
Fuses:
Ambient temperature:
Relative humidity:
Overvoltage category:
Pollution degree:
(See enclosed manual)

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3. Safety Notes
Caution!
The microscope’s electrical accessory com-
ponents are not protected against water.
Water 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
absolutely certain to switch off the main
power switch and remove the power cable.

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4. Overview of the Instrument
4. Overview of the Leica DMI 6000 B
4.1 Specifications
Contrast Methods
Transmitted Light Axis
Incident Light Axis
Tube
•Transmitted light (DL): BF, DF, PH, DIC, Pol
•Incident light (IL): Fluo
•Combination (DL/IL): Fluo/DIC, Fluo/PH
•Intermediate pupil: IMC (integrated modulation contrast)
IPH (integrated pos./neg. phase contrast)
•Automatic Illumination Manager
(aperture, field diaphragm, intensity, process switching)
•Automatic, color-neutral intensity control
•Manual or motorized shutter
•Lamp housing mount for interchangeable lamp housings.
•Automatic, electronic condenser identification
•Aautomatic Illumination Manager
(aperture, field diaphragm, intensity, process switching)
•Automatic, color-neutral intensity control
•Motorized shutter (switching speed < 50ms)
•Lamp housing mount for up to 3 interchangeable light sources
•Motorized 6-place filter turret
•Fluorescence Intensity Manager (FIM)
(reduction of incident illumination intensity)
•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
•Ultrafast filter wheel for 3 excitation wavelengths
(switching speed < 50 ms)
•Ergonomic with or without camera port at left
•2switching positions: 100%VIS and 50%VIS / 50%CAM
•Optional Bertrand lens
•Eye spacing adjustment
•Height and angle adjustment (10°- 45°)

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4. Overview of the Instrument
Magnification Changer
Objective Turret
Stages
•Motorized
•3switching positions
(choice of magnifications: 1x, 1.5x, 1.6x or 2.0x)
•Effective for all camera ports
or
•Manual
•2switching positions
(choice of magnifications: 1x, 1.5x, 1.6x or 2.0x)
•Effective on tube port
•Motorized and coded
•6x for objectives with M25 thread and 45mm parfocal distance
•For DIC: motorized/coded Wollaston prism carousel
•Anti-vibration locking
Fixed regular stages
•Ceramic-coated stage plate (248mm x 204mm)
•Heated stage plate (from 3°C above room temperature to 60°C)
(248 x 212mm)
•Temperature-controlled stage plate (0°C to 60°C) (248mm x
212mm)
Fixed micromanipulation stages
•Ceramic-coated stage plate (248mm x 204/122mm)
•Heated stage plate (from 3°C above room temperature to 60°C)
(248mm x 204/122mm)
•Temperature-controlled stage plate (0°C to 60°C)
(248mm x 204/122mm)
Regular manual 3-plate cross-stage
•Positioning range: 83mm x 127mm
•20 optional inserts (standard, heating, cooling) for a variety
of applications, size of inserts:160mm x 110mm (compatible
with scanning stages)
Manual micromanipulation 3-plate cross-stage
•Positioning range: 40mm x 40mm
•3optional inserts for a variety of applications
Motorized micromanipulation 3-plate cross-stage
•Positioning range: 40mm x 40mm
Scanning stage IM 120 x 100 (motors on top)
•1mm, 2mm, 4mm spindle pitch
(higher resolution v. higher speed)
•20 optional inserts (standard, heating, cooling) for a variety
of applications, size of inserts:160mm x 110mm

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4. Overview of the Instrument
Condensers
Z focus
Observation Ports
Scanning stage IM 120 x 100 (motors on bottom)
•1mm, 2mm, 4mm spindle pitch
(higher resolution vs. higher speed)
•20 optional inserts (standard, heating, cooling) for a variety
of applications, size of inserts:160mm x 110mm
•Motorized and coded
•Motorized or manual aperture diaphragm
•Contrast methods: BF, DF, PH, DIC, Pol, IMC
•Automatic method switching
•Condenser turret with 7 positions for contrast methods
•2condenser housings (S1-S28 and S70)
•Condenser heads: S1/1.4 oil, S1/0.9 dry, S23/0.53, S28/0.55
•Condenser heads can be swung out
•Condenser 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
•Motorized and coded
•9mm travel (1mm below, 8mm above the stage)
•Maximum travel speed: 5mm/s
•5focus 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
•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

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4. Overview of the Instrument
Controls
Electronics Box
Leica CTR6000
Interfaces
Software Tools
•7fixed control buttons for illumination and apertures
•7variable function buttons behind the focus controls
•3fixed control buttons for focus steps
•2focus hand wheels
•7buttons for fluorescence cubes and shutters
•4buttons for magnification changers and ports
•SmartMove: ergonomic controller for x, y, z and 4 additional vari-
able function buttons
•Separate control unit for all motorized and electronic elements of
the microscope such as:
•Objective turret
•Ffocus
•Ports
•Magnification changer
•Fluorescence
•Condenser
•Motorized stages
With
•Power supply for 100W halogen lamp
•Power supply for SmartMove
•2x RS232C
•2x USB
•4x external/internal peripherals
•Leica Application Suite (LAS) for WindowsTM 2000, XP with plug-ins
For:
•Microscope and camera configuration
•Microscope and camera control
•Image acquisition

16
4. Overview of the Instrument
4.2 Glossary
The Stand
Four basic versions of the Leica DMI stand (DMI
3000 B, DMI 4000 B, DMI 5000 B, DMI 6000 B) are
available which can be combined into a wide
range of microscope variants.
The basic building blocks of the Leica
DMI6000 B stand are:
•DMI6000 B electronic stand
•Integrated fluorescence axis with motorized
filter cube changer (6 positions)
•With or without bottom port
•Lateral camera ports, 100%, 80%, or 50%
•Optional top port at the left side of the tube
•Integrated motorized tube lens changer
•Optional Bertrand lens
The individual variants and their components,
differences, and applications are described in
this manual. The function and performance of all
microscopy techniques and required accesso-
ries of the Leica DMI6000 B will be described in
detail in the section of this manual that covers
the operation of the microscope.
Tube
The tube and tube lens create the primary image
together with the objective.
The tubes are an integral part of the stand and
consist of a basic body and a binocular section.
The trinocular tube also features a photo/video
port. A switchable mirror diverts either 100% of
the light to the eyepieces or camera port, or
splits it, with 50% each going to the eyepieces
and camera port.
A Bertrand lens is also available as an option.
Eyepieces
The eyepieces create an enlarged, virtual image
of the actual intermediate image created by the
objective. The eyepiece serves as a magnifier in
this respect.
Intensity Controller
The stand contains a 12V 100 W transformer for
continuous regulation of the intensity via the in-
tensity controller.
The intensity can be adjusted using the controls
(1.6).
Focus Wheel
The focus wheel allows quick, precise focusing
of the microscopic image. Focusing is realized
by the vertical travel of the objective turret, with
a total range of 9mm.
Incident Light-Fluorescence Unit
The stand features an integrated fluorescence
axis and a motorized filter cube changer with
6positions.
Aperture Ddiaphragm
The aperture diaphragm determines the resolu-
tion, depth of field, and contrast of the micro-
scopic image. The best resolution is obtained
when the apertures of the objective and the
condenser are roughly the same.
Caution:
The aperture diaphragm in the illumination
light path is not for setting the image bright-
ness. This should be done only with the inten-
sity controls or neutral filters.

17
4. Overview of the Instrument
Condenser
The condenser is a lens system that gathers
light and projects it onto the specimen from
above. The condenser is designed for the utiliza-
tion of the numerical aperture in the objective.
Condenser Height Adjustment
The markings of the transmitted-light column indi-
cate the height to be set for the used condenser.
Stages and Accessories
The stage is designed to accommodate the
specimens to be observed. Mechanical and mo-
torized 3-plate cross-stages are available for the
Leica DMI 6000 B.
Motorized Objective Turret and Objectives
The motorized objective turret is designed to ac-
commodate the objectives. The L-objectives
with their long working distances especially
take into consideration the correction of varying
container bottom thicknesses.
All microscope objectives are usable, from
1.25:1 to 100 : 1 magnification. All objectives in
the Leica product range with a 25mm thread and
coverslip correction are compatible. For perfor-
mance data on Leica objectives, please refer to
the most current valid objective lists available
from your Leica representative.
Transmitted Light Illumination Unit
The transmitted light illumination unit consists of
an illumination carrier and the transmitted light
illumination column. The transmitted light illumi-
nation carrier does not contain a lamp housing,
but a filter module for two swing-in filters and,
depending on the condenser used, a field dia-
phragm.
Filter
The filters are generally used to enhance the
contrast of the specimen and are installed in the
illumination carrier. A selection of various filters
can be interchanged as required.
Field Diaphragm
The field diaphragm is used to realize Koehler il-
lumination.
Lamp Housing for Transmitted Light
The lamp housings 107/2 and 107 (both for 12V
100W halogen) are available for the Leica DMI
6000 B. For their descriptions and applications,
please refer to the section of this manual de-
scribing the use of the microscope.
The letter L indicates a lamp housing designed
for left-handed operation.
Lamp Housing for Incident Light
The 106 z L lamp housing (for halogen or xenon)
is available for the Leica DMI 6000 B. For their
descriptions and applications, please refer to
the section of this manual describing the use of
the microscope.
The letter L indicates a lamp housing designed
for left-handed operation.
Leica CTR6000 Electronics Box
The Leica CTR6000 electronics box contains the
power supply for the lamp and the circuit boards
required to control the motorized functions of
the stand.

18
4. Overview of the Instrument
Fig. 1 Leica DMI 6000 B left view
1Eyepiece
2Eyepiece tube
3Top port
4Intermediate pupil interface
5LeicaScreen
6Light intensity
7Field diaphragm
8TL/IL switching
9Aperture diaphragm
10 Focus wheel
11 Variable function buttons
12 Right side port
13 Booster lens
14 Lamp mount
15 Condenser head
16 Condenser base
17 Field diaphragm
18 Transmitted light lamp housing
19 DIC objective prism disk
1
2
3
4
5
678910111213
14
15
17
16
18

19
4. Overview of the Instrument
Fig. 2 Leica DMI 6000 B right view
1E-focus control buttons
2Focus wheel
3Variable function buttons
4Opener for drawer
5Drawer
6Right side port
7Analyzer slot
8Centering window
9Field diaphragm centering
10 Incident light lamp housing
11 Objective turret
12 Stage with attachable mechanical stage
12 3
4
5
678
12
910
11

20
4. Overview of the Instrument
Fig. 3 Leica DMI 6000 B front view
1LeicaScreen
2Front control panel
3Port switching
4Top port
5Manual transmitted light filters
6Field diaphragm centering
1
2
3
4
6
5