Leica M205 FA User Manual.pdf

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Leica M165 FC

Leica M205 FA

User Manual

General Instructions

Safety concept

Before using your microscope for the first time, please read the "Safety concept" brochure included with your instrument. It contains additional information about handling and care.

Use in clean rooms

The Leica M series can be used in clean rooms without any problems.

Cleaning

ρρ Do not use any unsuitable cleaning agents, chemicals or techniques for cleaning.

ρρ Never use chemicals to clean colored surfaces or accessories with rubberized parts. This could damage the surfaces, and specimens could be contaminated by abraded particles.

ρρ In most cases, we can provide special solutions on request. Some products can be modified, and we can offer other accessories for use in clean rooms.

Servicing

ρρ Repairs may only be carried out by Leica Microsystems-trained service technicians. Only original Leica Microsystems spare parts may be used.

Responsibilities of person in charge of instrument

ρρ Ensure that the Leica stereomicroscope is operated, maintained and repaired by authorized and trained personnel only.

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Important Safety Notes

User Manual

The individual modules of the Leica M stereomicroscopy series include an interactive CD-ROM with all relevant user manuals in 20 other languages. Keep it in a safe place, and readily accessible to the user. User manuals and updates are also available for you to download and print from our web site at www.stereomicroscopy.com.

This User Manual describes the special functions of the individual modules of the Leica M stereomicroscopy series and contains important instructions for their operational safety, maintenance, and accessories.

The "Safety concept" booklet contains additional safety information regarding the service work, requirements and the handling of the stereomicroscope, accessories and electrical accessories as well as general safety instructions.

You can combine individual system articles with articles from external suppliers (e.g. cold light sources, etc.). Please read the user manual and the safety requirements of the supplier.

Before installing, operating or using the instruments, read the user manuals listed above. In particular, please observe all safety instructions.

To maintain the unit in its original condition and to ensure safe operation, the user must follow the instructions and warnings contained in these user manuals.

Special manuals

Special manuals are provided for a number of accessories:

ρρ User Manual M2-216-1 for lamp housing with high-pressure mercury burner.

ρρ User Manual for ebq 100 isolated supply unit for 100W high-pressure mercury burners­ .

ρρ User Manual M2-267-1 for the motorized focus system.

ρρ User Manual M2-166-2 for FluoCombi III™.

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Symbols Used

Warning of a danger

This symbol indicates especially important information that must be read and

complied with.

Failure to comply can cause the following:

ρρ

Hazards to personnel

ρρ

Functional disturbances or damaged instru-

 

ments

Warning of hazardous electrical voltage

This symbol indicates especially important information that is mandatory to

read and observe.

Failure to comply can cause the following:

ρρ

Hazards to personnel

ρρ

Functional disturbances or damaged instru-

 

ments

Danger due to hot surface

This symbol warns against touching accessible hot surfaces, e.g. those of

light bulbs.

Important information

This symbol indicates additional information or explanations that intend to

provide clarity.

Explanatory notes

ρρ This symbol within the text stands for additional information and explanations.

Figures

(1)Numbers in parentheses within the descriptions relate to the figures and the items within those figures.

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Safety Instructions

Description

The individual modules fulfill the highest requirements for observation and documentation of Leica stereomicroscopes of the M series.

Intended use

Refer to "Safety Concept" booklet

Non-intended use

Refer to "Safety Concept" booklet

Never use M series microscopes or their components for surgical procedures (e.g. on the eye) unless they are specifically intended for that purpose.

The instruments and accessories described in this operating manual have been tested for safety and potential hazards. The responsible Leica affiliate must be consulted whenever the instrument is altered, modified or used in conjunction with non-Leica components that

are outside of the scope of this manual.

Unauthorized alterations to the instrument or noncompliant use shall void all rights to any warranty claims.

Place of use

Refer to "Safety Concept" booklet

ρρ Electrical components must be placed at least 10 cm away from the wall and from flammable substances.

ρρ Avoid large temperature fluctuations, direct sunlight and vibrations. These conditions can distort measurements and micrographic images.

ρρ In warm and warm-damp climatic zones, the individual components require special care in order to prevent the build-up of fungus.

Responsibilities of person in charge of instrument

Refer to "Safety Concept" booklet

Ensure that:

ρρ The M series stereomicroscopes and accessories are operated, maintained and repaired by authorized and trained personnel only.

ρρ All operators have read, understood and observe this User Manual, and particularly the safety instructions.

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Safety Instructions (cont'd.)

Repairs, service work

Refer to "Safety Concept" booklet

ρρ Only original Leica Microsystems spare parts may be used.

ρρ Before opening the instruments, switch off the power and unplug the power cable.

Touching the live circuit can cause injury.

Transport

ρρ Use the original packaging for shipping or transporting the individual modules of the Leica M stereomicroscopy series and the accessory components.

ρρ In order to prevent damage from vibrations, disassemble all moving parts that (according to the user manual) can be assembled and disassembled by the customer and pack them separately.

Integration in third-party products

Refer to "Safety Concept" booklet

Disposal

Refer to "Safety Concept" booklet

Legal requirements

Refer to "Safety Concept" booklet

EC Declaration of Conformity

ρρ Refer to "Safety Concept" booklet

Health risks

Workplaces with stereomicroscopes facilitate and improve the viewing task,

but they also impose high demands on the eyes and holding muscles of the user. Depending on the duration of uninterrupted work, asthenopia and musculoskeletal problems may occur. For

this reason, appropriate measures for reduction of the workload must be taken:

ρρ

Optimal arrangement of workplace, work

 

assignments and work flow (changing tasks

 

frequently).

ρρ

Thorough training of the personnel, giving

 

consideration to ergonomic and organiza-

 

tional aspects.

ρρ

The ergonomic design and construction

 

of the Leica M stereomicroscopy series are

 

intended to reduce the exertion of the user

 

to a minimum.

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Safety Instructions (cont'd.)

Direct contact with eyepieces is a potential transmission method for bacterial

and viral infections of the eye.

The risk can be kept to a minimum by using personal eyepieces for each individual or detachable eyecups.

Light sources: Safety regulations

Protective measures of the manufacturer:

ρρ UV protection screen in front of the specimen plane prevents the user from looking directly into the UV rays.

ρρ

Dummy filter carriers in the free positions

 

of the rapid filter changer prevent direct UV

 

radiation from reaching the eyes.

ρρ

UV filters are installed in the observation

 

beam paths to protect the eyes.

ρρ

The stray-light protection on the lamp

 

housing prevents irradiation of the hands.

Warning

UV radiation could damage the eyes. Therefore:

ρρ

Never look into the light spot on the speci-

 

men plane without a UV protection screen.

ρρ

Never look into the eyepieces if no excita-

 

tion filter is in the beam path.

ρρ

Fill empty filter positions with dummy filter

 

carriers (e.g. M165 FC).

ρρ

Do not select a white, strongly reflective

 

background for the specimen.

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Safety Instructions (cont'd.)

Supply unit

Always unplug the supply unit from the power supply:

ρρ When installing and disassembling the lamp housing

ρρ

Before opening the lamp housing

ρρ

When replacing the high-pressure mercury

 

lamp and other parts, such as the heat-

 

absorbing filter or the collector

ρρ

During maintenance work on the supply

 

unit

Lamp housing

Never open the lamp housing while the lamp is switched on. Risk of explosion,

UV exposure, blinding!

Before opening the lamp housing, allow it to cool off for at least 15 minutes.

Danger of explosion!

Never cover the air duct on the lamp housing. Danger of fire!

Mercury lamp

ρρ Follow the user manual and safety instructions provided by the lamp manufacturer, and, in particular, the section on how to proceed if a lamp breaks and releases mercury.

ρρ

For transport, remove the mercury lamp,

 

transport it in its original packaging and

 

protect moving parts in the lamp housing

 

using the transport anchors.

ρρ

When it has reached the end of its rated life

 

(follow the manufacturer's specifications

 

and the minute meter on the supply unit).

ρρ

To minimize the risk of explosion, replace a

 

discolored mercury lamp promptly.

ρρ

Leica Microsystems assumes no liability

 

for damage caused by exploding, incor-

 

rectly installed or improperly used mercury

 

lamps.

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Contents

General Instructions

2

Important Safety Notes

3

Symbols Used

4

Safety Instructions

5

 

 

The Leica M Series for Fluorescence

 

Congratulations!

13

A Step Towards Infinity

14

The Electronics: Comfort, Convenience and Safety

15

The Modular Design: Everything is Relative

16

Maximum Compatibility

17

On We Go

18

 

 

Assembly

 

Assembling the Focusing Column for MDG Bases

20

Assembling the Focusing Column with an Incident-light Base

21

Motorized Focus: Restricting the Travel Path

22

Optics Carrier

23

Tube

24

Eyepieces

25

Objective

26

UV Protection Screen – Assembly

27

Objective Nosepiece – Assembly

28

Objective Nosepiece – Adjusting Parfocality

29

Transmitted-light Base TL ST

30

FluoCombi III™ – Preparations

31

FluoCombi III™ – Assembly

32

FluoCombi III™ – Adjusting the Parfocality

34

FluoCombi III™ – Centering the Objectives

36

FluoCombi III™ – Filter and UV Protection Screen

37

Transmitted-light Base TL BFDF: Before First Use

38

Transmitted-light Base TL BFDF

39

TL RC™ / TL RCI™

40

IsoPro™ Manual Mechanical Stage: Assembly

41

Leica IsoPro™ Motorized Mechanical Stage: Assembly

44

Cables: Connections

48

Cables: Cable Duct

49

Cables: Diagram

50

Leica LED5000 MCI™

51

Leica LED5000 MCI™: Alternative Assembly

52

Leica LED5000 RL

53

Leica EL6000 – Safety Notes

54

Leica EL6000 – Preparations

57

Leica EL6000 – Connection to the Fluorescence Lamp Housing

59

Leica EL6000 – Replacing the Lamp

61

Leica EL6000 – Connection to the Fluorescence Lamp Housing (cont'd.)62

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Quick Start Guide

 

The Fastest Route to Success

64

Overview of an M Series Microscope

65

The Correct Interpupillary Distance

66

Using the Eyepieces

67

Focusing

68

Adjusting the Resistance of the Focus Drive

69

Changing Magnification (Zoom)

70

Ratchet Steps and Magnification Levels

71

Parfocality: More Comfort and Convenience for Your Work

72

Iris Diaphragm

73

 

 

Eyepieces

 

Magnification Factors of the Eyepieces

75

Health Notes

76

Dioptric Correction

77

Dioptric Correction and Parfocality

78

Graticules

79

 

 

Photography & Video

 

Photography & Video

81

Photo Tubes and C-mounts

82

Trinocular Video/Phototube 50%

83

Trinocular Video/Phototube 100%

84

 

 

 

 

Microscope Carrier

 

The Objective Nosepiece

86

 

 

Objectives andOptical Accessories

 

The Different Types of Objectives

88

 

 

Bases

 

Leica TL ST Transmitted-light Base: Controls

90

Leica TL ST Transmitted-light Base: Operation

91

Leica TL ST Transmitted-light Base: Changing Bulbs

92

Leica TL BFDF Transmitted-light Base: Controls

93

Leica TL BFDF Transmitted-light Base: Operation

94

Leica TL RC™ / TL RCI™: Controls

95

Leica TL RCI™: The Deflection Mirror

96

Leica TL RCI™: Color Intensity and Temperature

97

Leica TL RC™ / TL RCI™: Operation

98

Leica TL RCI™: Methods in Transmitted Light

99

Leica TL RCI™: Relief Images

100

Using Filters

102

Leica IsoPro™ (Non-motorized): Controls

103

Leica IsoPro™ (Motorized): Controls

104

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System Illumination

 

Leica LED5000 MCI™

106

Leica LED5000 RL

108

Leica EL6000 – About the Instrument

109

Leica EL 6000 – Operation

111

Leica EL 6000 – Troubleshooting

112

Cleaning and Maintenance

113

 

 

Fluorescence

 

Filter Changer

115

FIM – Fluorescence Intensity Manager

116

About Fluorescence Microscopy

117

Rapid Filter Changers and Filter Types

118

Simple filter holders

119

Equipping the Filter Changer

120

Observation without Fluorescence

121

Double Iris Aperture

122

Commissioning the Fluorescence System

123

 

 

Accessories

 

Leica SmartMove

125

Leica SmartTouch™

126

Leica PSC Controller

127

 

 

 

 

 

Dimensional Drawings

 

Leica M165 FC

129

Leica M205 FA

132

Leica LED5000 MCI

135

Leica LED5000 RL

136

MATS TL

137

 

 

Specifications

 

Leica M165 FC / Leica M205 FA

139

Leica EL6000 – Specifications

143

Leica TL ST Transmitted-light Base

145

Leica TL BFDF Transmitted-light Base

146

Leica TL RC™ / TL RCI™

147

Leica IsoPro™ Motorized XY Stage

148

 

 

Appendix

 

Calculating the Total Magnification and Field of View Diameter

150

Care, Maintenance, Contact Persons

151

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The Leica M Series for Fluorescence

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Congratulations!

Congratulations on obtaining your new Leica M series stereomicroscope. We are convinced it will exceed your expectations, as never before have we applied our decades of experience in the areas of optics, mechanical engineering and ergonomics in such an uncompromising manner.

The Leica M series embodies all the qualities you associate with the name Leica Microsystems: excellent objectives, high-quality engineering, and reliability. Furthermore, the modular design ensures that the M series adapts perfectly to your needs—no matter which accessories you require for your tasks.

The entire imaging system, including the zoom, objective and ErgoTube®, is apochromatically corrected with much technological effort. Contrast, sharpness, richness in detail, resolution, image and color fidelity are optimum. In addition, the patented illumination beam path guarantees at every zoomlevelthatlightutilizationisatamaximumandthatfluorescenceimages are intensely luminous on a jet black background.

Though the reliability and robustness of Leica stereomicroscopes is legendary, like any high-tech product, the Leica M series requires a certain degree of care and attention. Therefore, we recommend that you read this manual. It contains all the information you need regarding operation, safety and maintenance. Simply observing a few guidelines will ensure that even after years of intensive use, your stereomicroscope will continue to work as smoothly and reliably as on the very first day.

We wish you the best of success in your work—after all, you are now equipped with the best tool!

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A Step Towards Infinity

Ever since their introduction by Horatio S. Greenough, stereomicroscopes have worked according to the optical principles based primarily on Ernst Abbe's research. For over a century, ingenious optics designers and engineers have worked to push magnification, resolution and image fidelity to the limit permitted by optics.

In doing so, they have always been constrained by the interrelation between three factors: the higher a microscope's resolution, the lower the available working distance. If one increases the distance of the optical axes, the three-dimen- sional image seen by the observer becomes distorted—a sphere becomes an ellipse, a flat surface curves towards the observer.

Limits are made to be broken

The Leica M205 FA is the world's first fluorescence stereomicroscope with a zoom range of 20.5:1. This accomplishment, however, was not enough for Leica's engineers. With the new FusionOptic™ in the Leica M205 FA, they have succeeded in going yet another step beyond previous limits. In addition to the increase in magnification, the resolution, too, has been increased to up to 1050 lp / mm, which corresponds to a resolved structure size of 472 nm.

Of course, this increase in performance benefits your everyday work. Set up your specimens on the microscope table with comfortable freedom of movement and discover details in stereomicroscopy that you could never see before.

* Leica M205 FA with 1.0× planapochromat and 10× eyepieces

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The Electronics: Comfort, Convenience and Safety for Your Experiments

Never before have electronics been used as extensively in a Leica series as in the new M series lineup. Optics carrier, stand, base and illuminator are all connected using electrical contacts—which provides a number of advantages.

Reliability for your experiments

Exact reproducibility of results is an increasingly common demand, particularly in research and development. The continuous encoding captures parameters such as the magnification, the illumination, the position of the iris

diaphragm and more and transmits the information to the Leica LAS software. Thus you always know the conditions under which an image was acquired.

Fewer cables

A large part of the cables have been routed in the interior of the column. Data are also transferred via the interface between the column, the optics carrier and the carrier. The direct result is that you need fewer cables—this not only makes your workstation neater and more comfortable, it even makes it appear larger.

Contacts not only transmit data, but also supply the power.

Leica Application Suite (LAS) evaluates the transmitted data and can restore the test situation later.

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The Modular Design: Everything is Relative

The Leica M series provides maximum flexibility in choosing equipment, thanks primarily to the modular configuration and the compatibility that Leica has painstakingly maintained for decades. The optics carriers, eyepieces, bases and more can be combined in any way you choose, allowing you to create the microscope that best suits your needs.

Despite this, you will notice that the controls and individual components do not differ significantly. Whichever configuration you choose, you will quickly feel right at home.

Have a special request? Let us know!

Leica Microsystems enjoys an exceptional reputation when it comes to devising customerspecific solutions. If you have a special request that cannot be met with standard parts, contact your Leica consultant. We have a solution for every problem.

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Maximum Compatibility

Leica engineers were careful to ensure that the new Leica M series—like its predecessors— remains compatible with existing series. This means that objectives, bases, tubes and so on can be reused.

Objectives

All new objectives of the Leica M series are parfocal, meaning that when used with the objective nosepiece, they can be replaced while keeping the specimen in perfect focus.

If you prefer, you can continue to use the previous Leica objective series. In this case, parfocality is no longer guaranteed.

Tubes

The interface between the optics carrier and the tube has remained the same, so existing tubes fit the new M series. The new tubes are designed for eyepieces with field number 23, while the predecessor models were only designed for field number 21, resulting in a smaller object field.

Eyepieces

However, the new M-series eyepieces have an audible and tangible click to provide immediate feedback in case of accidental adjustment.

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On We Go

If your new Leica microscope has already been assembled and commissioned by your Leica consultant, click here to skip through the installation instructions and go directly to the Quick Start Guide on Page 63.

If, on the other hand, you are assembling the microscope yourself, continue with the "Assembly"­ chapter, which begins on Page 19.

For everything you need to know about the correct use of fluorescence-related parts, refer to Page 114.

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Assembly

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Assembling the Focusing Column for MDG Bases

The first step is to connect the focusing column

Assembling the column adapter

Assembling the focusing column

of the M series to the corresponding base.

1. Securely install the column adapter on the

2. Securely screw the focusing column to the

 

column using the four included screws.

base using the six included screws.

Tools used

 

 

ρρ Hex socket screwdriver, 3 mm

 

 

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Assembling the Focusing Column with an Incident-light Base

When using an incident-light base, the focus-

Assembly

3. Screw the base securely onto the focusing

ing column and motorized focus are installed

1. Place the focusing column on the side.

drive.

directly on the base; no extension plate is

 

 

required.

 

 

Tools used

 

 

ρρ Hex socket screwdriver, 3 mm

 

 

2.Insert the four screws provided into the outer holes of the base.

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Motorized Focus: Restricting the Travel Path

Depending on the work situation, it is useful to restrict the maximum travel path of the stereomicroscope. This prevents

injury during handling of the sample caused by fingers or hands becoming trapped or the specimen touching the objective or even being damaged by it.

Readjusting the motorized focus

The motorized focus is factory-adjusted and normally does not need to be readjusted— even if the maximum travel path is changed.

Exception: If the power fails while the motorized focus is moving, the position

data are lost. In this case, the calibration must be repeated using the Leica LAS software or the Leica SmartTouch™. To do so, please consult the respective manual.

Restricting the bottom travel range

1.Move the motorized focus into the lowest position you want to reach.

2.Unscrew the screw of the limit stop on the side of the focusing column.

3.Push the limit stop to the height of the motorized focus.

It is easiest to move the limit stop by keeping the screwdriver inserted and moving it upwards.

4. Tighten the screw of the limit stop.

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Optics Carrier

Tools used

Assembling the optics carrier

ρρ Hex socket screwdriver, 4 mm

1. Place the optics carrier on the focusing

 

column so that the screw fits into the thread

 

provided and the lug fits into the groove.

2.Press the optics carrier backwards to the focusing column and screw it in place using your other hand.

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Tube

All intermediate tubes that fit between the optics carrier and the binocular tube are fitted in the same manner.

Tools used

ρρ No tools required.

Preparations

1.Unscrew the positioning screw and remove the protective cover.

Assembling the tube

2.Push the tube (for example, the inclined binocular tube) into the dovetail ring and rotate it slightly in both directions until the positioning screw meshes with the guide groove.

3.While holding the tube only slightly, carefully tighten the positioning screw. It is automatically brought to the correct position.

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Eyepieces

Tools used

Preparation

Inserting the eyepieces

ρρ No tools required.

1.

If you want to use an optional graticule,

3. Push the eyepieces into the tubes as far as

 

 

insert it now (Page 79).

they will go and check to ensure that they

Magnification range

 

 

fit tightly and accurately.

You can extend the overall magnification range

2.

Remove the plastic tube guard.

 

using available 10×, 16×, 25× and 40× wide-

 

 

 

field eyepieces for persons wearing glasses.

 

 

 

4. Securely tighten the clamping screws.

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Objective

Tools used

ρρ No tools required.

Preparation

1.Remove the protective cap on the optics carrier by turning it.

Attaching the objective

Hold the objective firmly during assembly and disassembly so that it does not

fall onto the stage plate. This applies particularly to the 2x planapochromatic objective, which is very heavy. Remove all specimens from the stage plate first.

2.Screw the objective clockwise into the optics carrier.

Alternative fastening options

ρρ If using the objective nosepiece, read the instructions on Page 28.

ρρ If using the Leica FluoCombi III™, read the instructions on Page 31.

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UV Protection Screen: Assembly

Tools used

ρρ Allen key

Intended use

The UV protection screen in front of the specimen plane prevents the user from looking directly into the UV rays.

Safety Notes

UV radiation can damage the eyes. Therefore, it is mandatory to install the

UV filter and adjust it correctly.

Always position the UV protection screen so that the operator can never

look directly at the light spot.

Use with one objective

1.Adjust the UV protection screen laterally using the arm.

2.Unscrew the hexagon-head screw.

3.Adjust the UV protection screen using the arm.

4.Tighten the hexagon-head screw.

5.Fasten the UV protection screen using a hexagon-head screw to the left or right side of the microscope carrier.

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Objective Nosepiece – Assembly

Preparations

Hold the objectives firmly during assembly and disassembly so that they

do not fall onto the stage plate.

ρρ Move the drive housing all the way upwards and remove the optics carrier, if the carrier has already been installed.

Assembly

1.Remove the transport anchor from the objective nosepiece.

2.Rotate the moving part by 90° and attach the objective nosepiece to the drive housing from the front (!). Screw the objective nosepiece firmly into place.

3.Unscrew the three Phillips screws on the objective mount of the optics carrier and remove the intermediate ring.

4.Screw the optics carrier onto the objective nosepiece.

5.Screw both objectives onto the objective nosepiece. It makes no difference which position an objective occupies.

6.Unscrew the locking screws on both sides of the objective nosepiece.

You can now adjust the parfocality (see instructions on the next page).

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Objective Nosepiece – Adjusting Parfocality

The following procedure only has to be carried out once. Afterwards, both objectives are parfocal, meaning that the specimen remains in focus when the objective nosepiece is rotated.

This procedure must be repeated if you replace either of the two objectives with another.

The following example assumes the combination of the 1× and 2× planapochromats. If you are using another objective combination, replace the 2× objective in the description with the objective with the stronger magnification.

Preparation

ρρ

Open the iris diaphragm.

ρρ

Set the dioptric correction of the eyepieces

 

to "0".

Adjustment

1.Rotate the 2× objective into the beam path and set it to the lowest magnification.

2.Focus on the specimen.

3.Rotate the 1× objective into the beam path.

4.Turn the objective on the thread in both directions until the specimen appears sharp.

5.Toggle to the 2× objective.

6.Select the strongest magnification and refocus until the specimen appears absolutely sharp.

7.Toggle to the 1× objective.

8.Turn the objective on the thread in both directions until the specimen appears absolutely sharp.

By means of zooming, check that the behavior of the objective is parfocal. Repeat the check with the other objective. If it is not parfocal, repeat the procedure.

9. Tighten the locking screws.

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Transmitted-light Base TL ST

Unpacking the base

Focusing drive and column

2.

Attach your focusing drive column from

The base is delivered with the adapter plate

1. Unscrew the extension plate from the base

 

below using the four hexagon-head

installed. Make sure the instruments are

using the Allen key provided.

 

screws.

unpacked on a flat, sufficiently dimensioned,

 

 

 

and non-slip surface.

 

3.

Re-attach the adapter plate to its origi-

 

 

 

nal position using the six hexagon-head

 

 

 

screws.

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FluoCombi III™ – Preparations

Before assembling the FluoCombi™, you have to disconnect all instruments from the power supply. Failure to observe these

instructions can result in damage to the microscope and the connected instruments.

Preparations

1.Remove the two screws used as transport anchors.

2.Screw the two screws into the holes provided for storage.

In this way, you can secure the FluoCombi III™ for transport whenever necessary.

3.Screw the 5× planapochromat into the FluoCombi III™.

4.Screw the other planapochromatic stereo objective into the FluoCombi III™.

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FluoCombi III™ – Assembly

1.Guide the FluoCombi III™ in from below, while holding it flat. Ensure that it is aligned with the holder on the focusing column at the left and right.

2.Hold the FluoCombi III™ securely and screw it to the focusing column using the provided screw.

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FluoCombi III™ – Assembly (cont'd.)

Assembling the optics carrier

1.Unscrew the three screws and the adapter ring on the optics carrier.

2.Attach the optics carrier to the FluoCombi III and screw it into place.

3.Open the iris diaphragm completely.

4.Assemble the tube (see Page 24).

5.Assemble the illuminator (see Page 51 ff.)

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FluoCombi III™ – Adjusting the Parfocality

Setting up the objective nosepiece

1.

Place a specimen under the microscope.

7. Unscrew

the clamping screw on the

The following configuration only has to

 

 

objective­

.

be carried out once. It ensures that the

2.

Rotate the stereo planachromatic objective

 

 

parfocality remains intact, and thus the speci-

 

under the beam path.

 

 

men does not need to be brought into focus

 

 

 

 

again.

3.

Reduce the view so that you can see the

 

 

 

 

specimen in the overview.

 

 

 

4.

Center the specimen under the beam path.

 

 

 

5.

Adjust the magnification to the highest

 

 

 

 

level and focus on a flat specimen.

 

 

6.Rotate the 5× planachromat under the beam path.

8.Reduce the view so that you can see the specimen in the overview.

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FluoCombi III™ – Adjusting the Parfocality (cont'd.)

9.By turning the objective ring, focus on the specimen until the specimen appears absolutely sharp.

10.Enlarge the view just enough for the center of the specimen to be visible in the field of view.

11.Refocus.

12.Tighten the clamping screw (see Step 7).

13.Rotate the stereo planachromatic objective under the beam path.

This causes the two objectives to be parfocal. This means that from then on, you can toggle between the two objectives without ­refocusing.

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FluoCombi III™ – Centering the Objectives

The following configuration only has to be carried out once. It ensures that the specimen remains in the optical center when

the objective is changed.

To attain optimum results for the follow- ing settings, insert a graticule into the eyepiece. This makes locating the center vastly

easier.

Centering the objective

1.Rotate the 5× planachromat under the beam path.

2.Focus on the midpoint of the specimen.

3.Rotate the stereo planachromatic objective under the beam path.

4.Center the objective over the specimen by loosening or tightening the screw on one side of the objective holder while turning the screw on the other side in the opposite direction.

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FluoCombi III™ – Filter and UV Protection Screen

Inserting the dichroic mirror

1.Pull the dummy filter carrier out of the holder.

Assembling the UV protection screen

1.Screw the UV protection screen to the holder provided on the optics carrier.

2. Insert the corresponding dichroic mirror.

2.Orient the UV protection screen so that you cannot look directly into the UV light.

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Transmitted-light Base TL BFDF: Before First Use

Removing the transport anchors

 

Before you can use the transmitted-

 

light base for the first time, it is abso-

 

lutely necessary to remove the two transport

Anchor of mirror

anchors.

 

Anchor of switching slide

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Transmitted-light Base TL BFDF

Standard delivery

The base is delivered with the adapter plate installed. The selected stage (Leica IsoPro™ mechanical stage or Leica standard stage 10 447 269), and the focusing drive will have to be mounted later.

Stage assembly

The Leica TL BFDF transmitted-light base can be equipped with three different stages.

ρρ

Leica IsoPro™ manual mechanical stage

ρρ

Leica IsoPro™ automatic mechanical stage

ρρ

Leica standard stage 10 447 269

The selected stage is mounted on the base before commissioning. You can switch between the stages at any time with just a few hand movements.

The following paragraph assumes use of the base without the stage mounted. Disassembly is performed in reverse order of the following steps.

Standard stage

1.Take the glass plate from the rectangular gap in the standard stage.

2.Position the stage on the transmitted-light base in such way that the four holes align over those in the base.

3.Attach the stage to the base with the four supplied hexagon-head screws.

4.Insert the glass plate back into the standard stage.

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TL RC™ / TL RCI™

The base is delivered with the adapter plate installed. The selected stage (Leica IsoPro™ mechanical stage or Leica standard stage 10 447 269), and the focusing drive will have to be mounted later.

Ensure that the instruments are unpacked on a level, adequately sized, and nonskid underlay.

Stage assembly

The Leica TL RC™/ RCI™ transmitted-light base can be equipped with three different Leica stages. The selected stage is mounted on the base before commissioning. You can switch between the stages at any time with just a few hand movements.

The following paragraph assumes use of the base without the stage mounted. Disassembly is performed in reverse order of the following steps.

Standard stage

1.Take the glass plate from the rectangular gap in the standard stage.

2.Position the stage on the transmitted-light base in such way that the four holes align over those in the base.

3.Attach the stage to the base with the four supplied hexagon-head screws.

4.Insert the glass plate back into the standard stage.

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IsoPro™ Manual Mechanical Stage: Assembly

Leica IsoPro™ Mechanical Stage

Before the Leica IsoPro™ mechanical stage is mounted to the base, the axis containing the control buttons is attached either on the left or the right side of the mechanical stage.

If the controls are to be mounted on the lefthand side, the gear rod on the bottom side of the mechanical stage must be unscrewed and reattached in reverse.

1.Take the glass plate from the mechanical stage.

2.Turn the mechanical stage around and place it onto a non-slip surface.

3.Change the gear rod from the left to the right-hand side.

4.Skip the next two steps to install the controls.

Before the Leica IsoPro™ mechanical stage is mounted to the base, the axis containing the control buttons is attached either on the left or the right side of the mechanical stage.

Left or right operation

If the controls are to be mounted on the lefthand side, the gear rod on the bottom side of the mechanical stage must be unscrewed and reattached in reverse.

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Leica IsoPro™ Manual Mechanical Stage: Assembly (cont'd.)

1.Take the glass plate from the mechanical stage and turn it around.

2.Change the gear rod from the left to the right-hand side.

Control assembly

1.Take the glass plate from the mechanical stage and turn it around.

2.Attach the axis with the control buttons to the desired side. The fastener snaps into the mechanical stage magnetically.

4.Attach the axis with the two supplied hexa- gon-head screws.

5.Attach the cover rail to the mechanical stage.

Mechanical stage assembly

1.Place the mechanical stage onto the base.

2.Pull the upper part of the mechanical stage carefully towards the user, fixing the lower part onto the transmitted-light base.

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Leica IsoPro™ Manual Mechanical Stage: Assembly (cont'd.)

3.Attach the mechanical stage evenly to the three threaded holes.

4.Now move the mechanical stage as far as it will go in the direction of the column.

5.Insert the glass plate into the mechanical stage.

Focusing drive and column

1.Unscrew the extension plate from the base using the Allen key provided.

2.Attach your focusing drive column from below using the three hexagon-head screws.

3.Re-attach the adapter plate to its original position using the three hexagon-head screws.

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Leica IsoPro™ Motorized Mechanical Stage: Assembly

Basics

The transmitted light bases of the Leica TL series (TL BFDF, TL RC™, TL RCI™) are supplied with an installed extension plate. The selected stage (Leica IsoPro™ mechanical stage or standard stage 10 447 269) and the focusing drive will have to be mounted later.

The motorized mechanical stage is a sensitive precision instrument. During installation, avoid subjecting the stage to impact or severe vibrations.

1.Unpack the mechanical stage from the transport packaging and position it on the transmitted-light base.

1

2

2.Secure the motorized mechanical stage to the base using three M4 screws.

1

Microscope base

3× M4 screws

2

Motorized mechanical stage

 

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Leica IsoPro™ Motorized Mechanical Stage: Assembly (cont'd.)

3.Remove the two M3 screws and the holder from the mechanical stage.

4.Remove the two M4 screws and the sleeve from the mechanical stage.

5.Remove the four shock-absorbing cartons from the mechanical stage.

After removal, keep all the transport anchors in the plastic bag provided for

future transport.

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Leica IsoPro™ Motorized Mechanical Stage: Assembly (cont'd.)

The bases

The TL BFDF, TL RC™ and RCI™ transmitted-light bases can be equipped with three different stages: Standard stage, manual and automated IsoPro™ mechanical stage. The selected stage is mounted on the base before commissioning. You can switch between the stages at any time with just a few hand movements.

The following paragraph assumes use of the base without the stage mounted. Disassembly is performed in reverse order of the following steps.

Mechanical stage and base

Never move the sledge of the motorized mechanical stage manually in the

X direction, as otherwise the mechanical system will be damaged!

1.Place the mechanical stage onto the base.

2.Pull the upper part of the mechanical stage carefully towards the user and fasten the lower part onto the transmitted-light base.

3.Attach the mechanical stage evenly to the three threaded holes.

4.Now move the mechanical stage as far as it will go in the direction of the column.

5.Insert the glass plate into the mechanical stage.

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Leica IsoPro™ Motorized Mechanical Stage: Assembly (cont'd.)

Mechanical stage for X-Y stage control module

1.Plug the CTL2 plug of the mechanical stage motor into one of the three available CTL2 interfaces.

2.Plug the CTL2 plug of the Leica PSC controller into another CTL2 interface.

ρρ The third CTL2 interface is available for connecting other instruments, such as the Leica foot switch (10 447 398) or TL RCI™ transmitted light base (10 446 352).

ρρ The 15-pin Sub-D interface is intended for use with the Leica SmartMove™ controller (11 501 197).

3.Plug a power cable into the socket provided and into a grounded power socket.

As soon as all instruments have been plugged into the control box and it has been plugged into the power supply, the mechanical stage initializes and automatically drives to the mid position.

4.Connect the PC (where applicable) and X/Y Stage DCI Module (using a suitable USB cable).

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Cables: Connections

The new Leica M-series features extensive encoding with which various microscope data and settings can be read out, transferred to the PC and reproduced later.

ρρ The interface to the optics carrier is on the column.

The terminals

1

3

2

The connection to the PC and to other instruments is made using the terminals on the rear side of the column:

1.2 CTL2 terminals for auxiliary equipment such as the ring illuminator, the base and other accessories from the Leica product range.

2.Terminal for the 50-watt power supply provided.

3.USB terminal for the connection to the PC.

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Cables: Cable Duct

The integrated cable duct in the column enables a neat cable layout around the microscope. For example, the USB or FireWire cables of the camera can be stowed in the cable duct.

Feeding the cables

1.Unscrew the three screws on the cable duct.

2. Remove the cover of the cable duct.

3.Place the cables in the cable duct and screw the cover on tightly.

Tip: Estimate the length of the cable ends you will need before screwing on the cover. For thick cables, it is difficult to change the length retroactively.

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Cables: Diagram

Alternative installation

Please use the Leica Application Suite to configure the functions of the system.

LED5000 RL

LED5000 MCI™

DCI Controlbox

Power/USB

115/230 V

TL RCI™

PC USB

 

Footswitch(es)

Smart Move

PSC

SmartTouch

Cross Stage

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Leica LED5000 MCI™

The Leica LED5000 MCI™ (for "Multi Contrast

Assembly

2. Connect the CAN-bus cable provided to

Illumination") is installed using two screws. For

1. Hold the LED5000 MCI™ with one

either of the two sockets. (The flat part of

optimum accessibility, the optics carrier should

hand and tightly screw the retain-

the plug must be facing downwards.)

be removed during assembly (see Page 23.)

ing stirrups on both top holes on the drive

 

 

­housing.

 

3. Plug the other end of the cable into one of the two "CTL2" sockets on the column.

Constraints

The Leica LED5000 MCI™ cannot be used together with the objective nosepiece.

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Leica LED5000 MCI™: Alternative Assembly

Under certain circumstances, the light source must not be moved along with the optics carrier. A typical example is multifocus images in which the Z-stack changes while the angle of incidence of the light must remain the same. For such purposes, the Leica LED5000 MCI™ is directly fastened to the column.

Installation on the column

1.Pull the retaining stirrup out of the Leica LED5000 MCI™.

2.Screw the retaining stirrup into the column at the notch using the single screw. If you

tighten the screw, the retaining stirrup is automatically moved into the correct position­ .

3.Push the Leica LED5000 MCI™ onto the retaining stirrup.

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Leica LED5000 RL

The ring illuminator, the Leica LED5000 RL ("Ring Light"), is installed on the objective using a single screw. It has been optimized for a working distance between 60 and 70 mm.

Constraints

The Leica LED5000 RL can be used in conjunction with the planapochromat 1× and planapochromat 0.63× objectives. With all other objectives, the working distance is too low for adequate illumination.

The ring illuminator cannot be used together with the objective nosepiece.

Assembly

1.Connect the CAN-bus cable to the ring illuminator. The flat part of the plug must be facing upwards.

2.Push the ring illuminator over the objective as far as it will go and screw it into place.

3.Plug the other end of the cable into one of the two "CTL2" sockets on the column.

For optimum accessibility of the specimen, the ring illuminator should be installed with the cable facing backwards. However, it is also possible to turn the ring illuminator sideways, for example if simultaneously using the Leica LED5000 MCI™ system illumination. In this case, the ring illuminator cannot be connected directly to the Leica LED5000 MCI™.

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Leica EL6000 – Safety Notes

The manufacturer assumes no responsibility or liability for any use outside of

the intended use or use outside of the specifications from Leica Microsystems Wetzlar GmbH or any risks resulting from such use.

In such cases, the Declaration of Conformity shall be invalid.

This (IVD) device is not intended for use in the patient environment defined by

DIN VDE 0100-710. It is also not intended to be combined with medical devices as defined by EN 60601-1. If a microscope is electrically connected to a medical instrument in accordance with EN 60601-1, the requirements defined in EN 60601-1-1 shall apply.

General safety notes

This safety class 1 device was built and tested in accordance with

EN 61010-2-101:2002 EN 61010-1:2001, IEC 1010-1:2001,

Safety requirements for electrical equipment for measurement, control and laboratory use.

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 conjunction with non-Leica components that are outside of the scope of this manual.

Unauthorized alterations to the instrument or noncompliant use shall void all

rights to any warranty claims.

If any safety defects or malfunctions of the Leica EL6000 are identified, the

instrument must be immediately disconnected from the power system and secured against additional use. For repair, the Leica EL6000 must be sent to the supplier or an authorized representative of the supplier.

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Leica EL6000 – Safety Notes (cont'd.)

The Leica EL6000 compact light source generates high-energy light with invisi-

ble components. There is a risk of being dazzled or blinded by light! Never look directly into the light guide output of the instrument or into the output of the light guide connected to the instrument. Always ensure that the output of the connected light guide is securely connected to the system to be illuminated before switching on the compact light source. Furthermore, before switching on the compact light source, dim the light output all the way using the intensity switch (left limit stop of the switch) to prevent damage to the connected system.

To prevent damage to the light guide connected to the device, the heat-

absorbing filter inserted into the device must be free of damage (gaps, cracks etc.). If the heat-absorbing filter is not inserted or is defective, the service life of the light guide will be decreased.

The interior of the lamp inserted into the unit contains highly toxic substances.

At the end of its service life, dispose of the lamp according to applicable regulations. The manufacturer of the lamp provides corresponding information, for example in the lamp's shipping documents. Follow the manufacturer's instructions in case the lamp bursts.

The Leica EL6000 compact light source is designed for a voltage range of

100 – 240 VAC, 50 – 60 Hz. Within this voltage and frequency range, the instrument always adapts to the connected power supply. Operating the instrument with a power supply voltage outside this range can destroy the instrument and the connected components!

Only fuses of the specified type and the listed rated current may be used as

replacements. The use of incorrect fuses may result in a fire hazard.

The instrument is intended exclusively for operation in dry rooms. Do not

use the instrument in rooms with explosion hazard.

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Leica EL6000 – Safety Notes (cont'd.)

Electrical safety

For indoor use only.

Supply voltage

100–240 V~ (±10%)

 

 

Frequency

50–60 Hz

 

 

Power consumption

Max. 210 VA

 

 

Fuses

5×20, 2.5 A, slow-blowing, breaking capacity H, see Page 113

 

 

Ambient temperature

0 – 40 °C

 

 

Relative humidity

10%–90%

 

 

Overvoltage category

II

 

 

Pollution degree

2

 

 

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Leica EL6000 – Preparations

Setting up the instrument

Set up the Leica EL6000 so that the front is readily accessible and visible.

ρρ

The ventilation slots on the sides and back

 

panel of the device must not be covered.

ρρ

Maintain a free room of at least 150 mm on

 

the rear panel of the instrument.

Instructions for lamp replacement

The lamp contains mercury! Be absolutely certain to follow the handling

instructions and safety notes provided with the lamp.

Maintain cleanliness when inserting the lamp. Remove any clinging packaging

materials.

The lamp used in the instrument becomes very hot during operation and

has high internal pressure when hot. Before changing the lamp, it is mandatory to allow the instrument to cool off for at least 20 minutes.

The Leica EL6000 compact light source comes with a factory-supplied lamp,

but the lamp is not installed when shipped. This minimizes the danger of damage to the lamp during transport.

Before transporting the Leica EL6000 compact light source, you first have to

remove the lamp.

Inserting the lamp

1.Pull the power plug out of the socket, so that the EL6000 is disconnected from the power system.

2.Unscrew both screws of the cover using a 3 mm Allen key.

3. Remove the housing cover.

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Leica EL6000 – Preparation (cont'd.)

4.Carefully lay the Leica EL6000 on its side so that the opening faces you.

5.Pull the pressure bolt back towards the front plate using the lever.

Ensure that the groove in the contact surface of the lamp matches up with the corresponding plug in the lamp mount.

This makes it easier to insert the lamp.

6. Insert the lamp.

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Leica EL6000 – Connection to the Fluorescence Lamp Housing

Always connect the light guide to the lamp housing of the stereomicroscope

in order to prevent danger to the user from the high-energy light.

1.Unscrew the positioning screw and remove the protective cover of the fluorescence lamp housing.

2.Insert the adapter into the fluorescence lamp housing and retighten the locking screw.

3.Unscrew the three red protective caps on the light guide.

4.Insert the short end of the light guide into the lamp housing of the stereomicroscope as far as it will go.

5.Securely tighten the clamping screw.

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Leica EL6000 – Preparation (cont'd.)

7.When the lamp makes full contact with the lamp mount, release the lever.

8.Connect the plug of the lamp with the coupling in the instrument.

9.Check to make certain that a heat-absorb- ing filter has been inserted into the shaft in order to protect the connected light guide.

Ensure that no wires are touching the reflector of the lamp.

10.Close the housing cover and insert the screws.

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Leica EL6000 – Replacing the Lamp

The procedure for replacing the lamp does not differ from the installation procedure outlined on Page 57, except for the fact that the existing lamp has to be removed first.

Replacing the lamp

The lamp used in the instrument becomes very hot during operation and has high internal pressure when hot. Before changing the lamp, it is mandatory to allow the

instrument to cool off for at least 20 minutes.

1.Pull the power plug out of the socket, so that the EL6000 is disconnected from the power system.

2.Unscrew both screws of the cover using a 3 mm Allen key.

3.Remove the housing cover.

4.Carefully lay the Leica EL6000 on its side so that the opening faces you.

This makes it easier to insert the lamp.

5.Pull the pressure bolt back towards the front plate using the lever.

6.Pull out the lamp and disconnect the plug from the coupling in the instrument.

7.Install the new lamp as outlined on Page 58, beginning with item 6.

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Leica EL6000 – Connection to the Fluorescence Lamp Housing (cont'd.)

6.Insert the long end of the light guide into the light output of the Leica EL6000. There must be a noticeable click.

Only use a light guide with a light input of the type "Storz long", as otherwise

damage to the instrument and danger to the user can result (risk of being dazzled).

Before you open the shutter, the light guide must be connected on both

sides. Otherwise, the escaping light can cause injury to eyes and skin and damage to furniture. Never look into the light escaping from the light guide!

5.Connect the Leica LA6000 to the power system using the power cable.

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Quick Start Guide

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The Fastest Route to Success

Your Leica stereomicroscope has been delivered in completely assembled condition by your Leica partner, and naturally you want to get right to work. Therefore, your next step should be to study the Quick Start Guide, which outlines the most important steps at a glance.

This manual will then familiarize you with the finer details of your microscope. The following pages contain important, practical information that makes using it every day easier.

Take time to read it – it's worth it!

The M series Quick Start Guide (included with instrument).

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Overview of an M Series Microscope

1Eyepieces for spectacle wearers with dioptric correction and eyecups

2Trinocular tube

3Fluorescence housing

4Filter Changer

5Display

6Motorized Focus

7Objective Nosepiece

8Leica SmartTouch

9UV protection screen

1

2

3

4

5

6

7

8

9

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The Correct Interpupillary Distance

The interpupillary distance is correctly set if you see a single circular image field when looking at a specimen.

If you are still a novice microscope user, you may need a short time to become accustomed to this. Not to worry—after a little while, it will become automatic.

Reference value

The distance between eye and eyepiece measures approx. 22 mm for 10/23B wide-field eyepieces for persons wearing glasses.

Adjusting the interpupillary distance

1.Look into the eyepieces.

2.Hold the eyepieces with both hands. Push the eyepieces together or separate them until you see a circular image.

3.Slowly approach the eyepieces with your eyes until you can see the complete image field without corner cutting.

 

 

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Using the Eyepieces

The eyepieces form the connection between the tube and the eye of the observer. Simply push them into the tube and they are ready to use.

Each eyepiece offers a certain magnification factor that has a determinative effect on the total magnification. Furthermore, all Leica eyepieces can be equipped with practical graticules that enable measuring and quantifying of specimens.

Dioptric correction

A built-in dioptric correction is available for eyeglass wearers. For more information, refer to Page 77.

If you do not wear glasses:

1.Hold the eyepiece firmly and rotate the eyecups forwards counterclockwise.

2.If an eyepiece is equipped with the inte-

grated dioptric correction, turn the value to the "0" mark.

If you wear glasses:

1.Hold the eyepiece firmly and rotate the eyecups clockwise towards the rear, as otherwise the viewing distance is too great.

2.If an eyepiece is equipped with the integrated dioptric correction, turn the value to the "0" mark.

By the way, one benefit of viewing with eyeglasses is a drastically lower risk of bacterial transmission (see Page 76). The soft material of the eyecup also ensures that your glasses will not be scratched, even if they contact the eyepiece.

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Focusing

Focusing raises or lowers the stereomicroscope using the focusing drive. The specimen detail is brought into sharp focus as soon as it is in the focal point of the objective.

ρρ The focusing drive can be operated either leftor right-handed.

Focusing

ρρ The inner, coarse adjustment is used for covering great distances.

ρρ The outer, fine adjustment is used for fine focusing.

Coarse/fine adjustment

The sharpness is adjusted using the coarse/fine adjustment. The resolution of the coarse/fine adjustment is 1 µm.

2. The coarse/fine adjustment carries a load of up to 15 kg.

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Adjusting the Resistance of the Focus Drive

Adjusting the resistance

Is the focus movement too loose or too tight? Does the outfit tend to slide downwards? The resistance can be adjusted individually depending on the equipment weight and personal preferences as follows:

1.Grip the outer drive knobs with both hands and turn them towards each other until the desired resistance is reached during focusing­ .

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Changing Magnification (Zoom)

All M series microscopes have an integrated zoom. The name indicates the zoom range covered:

ρρ

Leica M165 C = 16.5:1

ρρ

Leica M205 C = 20.5:1

The rotary knob for the zoom can be used either left or right-handed.

Zooming

1.Look into the eyepieces.

2.Focus on the specimen.

3.Rotate the magnification changer until the desired magnification is configured.

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Ratchet Steps and Magnification Levels

The zoom button can optionally be operated

Enabling and disabling ratchet steps

Magnifications and fields of view

either with or without ratchet steps. Continu-

1.

Push the top button downwards to disable

The formula on Page 147 provides additional

ous zoom is possible when the ratchet steps are

 

the ratchet steps.

information about the magnifications and field

disabled, which many users find convenient.

 

 

of view diameters, with consideration given to

On the other hand, when the ratchet steps are

2.

Push the bottom button upwards to enable

the position of the magnification changer and

enabled, photographs, measurement results

 

the ratchet steps.

the eyepiece and objective combination used.

etc. can be reproduced more accurately.

 

 

 

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Parfocality: More Comfort and Convenience for Your Work

All Leica stereomicroscopes are parfocally matched, meaning that you can view a focused specimen from the lowest to the highest magnification without having to refocus. The focus needs only be readjusted if you want to see a specimen location that is located higher or lower.

Requirements for parfocal work

ρρ If you are using an eyepiece with dioptric correction, the procedure differs from this description. For more information, refer to Page 78.

ρρ

For the procedure for adjusting the parfo-

 

cality for the objective nosepiece, refer to

 

Page 29.

ρρ

For the procedure for adjusting the parfo-

 

cality for the Leica FluoCombi III™, refer to

 

Page 34.

Parfocality

1.Enlarge the view to the maximum level.

2.Focus on the specimen.

You are done! Even if you select a smaller working distance, the specimen remains pin-sharp.

The parfocality is maintained until you focus on another level of the specimen.

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Iris Diaphragm

The iris diaphragm in the optics carrier of your M series microscope fulfills the same purpose as the iris diaphragm in a camera: it regulates the available light, which changes the depth of field. The "depth of field" (or "focus depth") is the area of a specimen that is brought into sharp focus.

Closing the iris diaphragm

ρρ Close the iris diaphragm by turning the knob to the left. The subject appears darker and the depth of field increases.

Opening the Iris Diaphragm

ρρ Open the iris diaphragm by turning the knob to the right. The subject now appears brighter, but the depth of field decreases.

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Eyepieces

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Magnification Factors of the Eyepieces

An eyepiece not only makes it possible to look passively into the microscope, but also has a critical effect on the maximum magnification. The magnification factor is between 10x and 40x.

The following eyepieces are available for the M series:

Magnification

Dioptric correction

Order number

 

 

 

10×

± 5 diopter settings

10 450 023

 

 

 

16×

± 5 diopter settings

10 450 024

 

 

 

25×

± 5 diopter settings

10 450 025

 

 

 

40×

± 5 diopter settings

10 450 026

 

 

 

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Health Notes

Potential sources of infection

Direct contact with eyepieces is a potential transmission method for

bacterial and viral infections of the eye. The risk can be kept to a minimum by using individual eyepieces or detachable eyecups. Eyecups can be ordered separately. Please contact your Leica partner.

Separate eyecups are an effective way of preventing infections.

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Dioptric Correction

All Leica eyepieces are also available with builtin dioptric correction, allowing the microscope to be used without glasses even by those with vision problems. The correction comprises ±5 diopter settings.

Using the Dioptric Correction

1.Set the dioptric correction of both eyepieces to the mid position ("0" diopter settings).

2.While wearing your glasses, look through the eyepieces and focus on the specimen.

3.Rotate both eyepieces to the maximum value of "+5".

4.Hold one eye closed and rotate the other eyepiece in the "-" direction until the specimen appears sharp.

5.Then, open the other eye and correct the diopter settings until the image is uniformly sharp.

Note that when using dioptric correction, the advantage of parfocality is lost—thus you have to manually refocus each time you

change the zoom level. To also use parfocality with dioptric correction, refer to the instructions on Page 78.

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Dioptric Correction and Parfocality

Leicastereomicroscopesareparfocallymatched. The prerequisite for this is the correct setting of the diopters and the parfocality. The following adjustments only have to be carried out once by each user.

Preparations

ρρ Move the lever of the video/phototube to the "observation" position and open the diaphragm.

ρρ If you are using the microscope carrier AX, set it to stereoscopic observation.

Adjusting

1.Set the dioptric correction for both eyepieces to "0".

2.Select the lowest magnification and focus on a flat specimen.

3.Select the highest magnification and readjust the sharpness.

4.Select the lowest magnification again, but do not look into the eyepieces.

6. Rotate the eyepieces counterclockwise in the "+" direction as far as they will go (+5 diopter settings).

7.Look into the eyepieces.

8.Slowly rotate each eyepiece individually in the "–" direction until each eye sees the object sharply imaged.

9.Select the highest magnification and refocus­ if necessary.

Now, if you adjust the magnification from the lowest to the highest level, the specimen is always brought into sharp focus. If not, repeat the process.

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Graticules

Use

Leica graticules make length measurements and counting easier, particularly for workstations that are not equipped with a digital camera and LAS software.

The Leica graticules for length measurements and numbering are fitted in mounts and are inserted into the eyepieces.

1. Screw the insert off of the eyepiece.

2.Clamp the graticule on the insert, applying moderate pressure. Ensure that the graticule fits tightly.

3.Screw the insert and graticule firmly into place and replace the eyepiece in the tube.

4.You can now align the graticule by rotating the eyepiece in the tube and then tightening it using the clamping screw.

Use with the Leica AX carrier

If possible, measure with the micro- scope carrier AX in vertical position. The measurements are more accurate without the

convergence angle in the stereoscopic image.

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Photography & Video

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Photography & Video

For most microscope users, digital documentation has become an invaluable part of their work. Research results can be presented in an attractive manner; measurements on the digital image provide clarity and, in conjunction with the motorized IsoPro™ mechanical stage, even images of large specimens can be captured step by step and automatically joined to create a new complete image.

Adapter

If camera control using the Leica Application Suite is not required, conventional mirror reflex and rangefinder cameras from third-party manufacturers can be used. For this purpose, Leica Microsystems offers a variety of adapters that can be used together with the 50% and 100% trinocular tubes.

Leica DFC cameras

However, if you require absolute control over the camera and need the capability for measurement, evaluation and more in addition to photography, the digital Leica DFC cameras are exactly right for you. Together with the Leica Application Suite, they provide virtually limitless freedom of use. For additional information about Leica cameras, refer to the camera's documentation.

Leica Application Suite

The "Leica Application Suite", or "LAS" for short, is, as it were, the digital extension of the Leica M series microscopes. In addition to capturing images, it lets you control the microscope, illuminator, stages, cameras and more. For additional information, refer to the LAS online help.

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Photo Tubes and C-mounts

Application

All Leica DFC cameras are equipped with a standardized C-mount interface. In turn, the C-mount adapter for the respective trinocular tube is connected to this interface. This adapter creates a solid mechanical connection between the microscope and camera and ensures optimum rendering of the microscopic image on the image sensor of the camera.

Usually, the ideal is for the digital camera to capture as much of the field of vision as possible, while excluding as much of the black edge of the field of vision as possible. To do so, the magnification factor of the C-mount adapter must match the image format of the sensor as closely as possible (see table).

If there is unwanted shading at the corners even with a compatible C-mount adapter installed, it can be corrected using the "Shading function" of the camera software.

Alternatively, you can also use a C-mount adapter with higher magnification. This primarily avoids the critical border area of the field of vision and concentrates on the center of the field of vision.

Camera

optimal

suitable

 

(large

(smaller

 

image field)

image field)

 

 

 

DFC290

0.5×

0.63×

 

 

 

DFC420

0.5×

0.63×

 

 

 

DFC490

0.63×

0.8×

 

 

 

DFC500

0.63×

0.8×

 

 

 

Cameras from third-party suppliers

In addition to Leica DFC cameras with the standardized C-mount interfaces, you can connect third-party cameras to the microscope using a T2 bayonet adapter. To do so, instead of the C-mount adapter, simply use the corresponding SLR adapter with T2 connection. However, these third-party cameras are not integrated into the Leica Application Suite and have to be operated using the corresponding software from the camera manufacturer.

The Leica digital cameras are detailed in a separate user manual along with instructions for their assembly and use.

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Trinocular Video/Phototube 50%

Use

With its third beam path, the trinocular video/ phototube 50% enables you to simultaneously view and photograph a specimen. The available light is divided as follows:

ρρ

50% is available for the two eyepieces.

ρρ

50% of the light is diverted to the video/

 

photo beam path.

Assembly

Fasten the "trinocular tube 50%" to the optics carrier instead of the binocular observation tube (refer to Page 24).

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Trinocular Video/Phototube 100%

Use

With its third beam path, the trinocular video/ phototube 100% enables you to either view or photograph a specimen. This means that 100% of the light is available to one or the other beam path. The other beam path remains opaque or black.

Assembly

Fasten the "trinocular tube 100%" to the optics carrier instead of the binocular observation tube (refer to Page 24).

Switchover

ρρ Turn the controller on the right side of the tube into the horizontal position in order to guide all available light into the eyepieces. You can now observe the specimen.

ρρ Turn the controller on the right side of the tube into the vertical position in order to guide all available light into the camera. You can now photograph the specimen.

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Microscope Carrier

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The Objective Nosepiece

Use

The objective nosepiece enables you to switch between two objectives using just one hand movement, for example between a 1× and a 2× planapochromat.

Parfocal work

With the new M series objectives, the parfocality is maintained even when objectives are changed, meaning that the specimen remains in sharp focus during the change.

Older Leica objectives can continue to be used, but without parfocality during the

objective change.

Technical constraints

The objective nosepiece cannot be used together with the LED5000 MCI™

system illumination, as the objective nosepiece can no longer be rotated.

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Objectives and

Optical Accessories

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The Different Types of Objectives

To meet the various requirements regarding imaging properties, there is a choice of highquality interchangeable planachromatic and planapochromatic objectives and also lowerpriced interchangeable achromatic objectives.

ρρ Achromatic objectives are particularly suited for specimens with high-contrast structures.

ρρ Flat-field (planachromatic) objectives are particularly well suited for studying flat objects such as wafers and thin sections.

ρρ With planapochromatic objectives, the finest structures are visible with high contrast. The sophisticated apochromatic correction allows these objectives to attain the highest color brilliance and fidelity.

Achromatic objectives

The 0.32×, 0.5×, 0.63×, 0.8×, 1×, 1.5×, 2× achromatic objectives offer countless variants for selecting the object field diameter, magnification ranges and working distances (see Page 147).

Planachromatic objective 1×

For the highest requirements for overall image quality, we recommend equipping the microscope with the 1× plan (flat-field) objective, which returns sharp, contrast-rich object fields.

Achromatic objectives with a long focal length

For special applications, achromatic objectives with long working distances and focal lengths of f=100 mm to 400 mm are available.

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Bases

 

 

 

 

 

 

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Leica TL ST Transmitted-light Base: Controls

 

1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

1

2

3

 

3

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

4

 

 

 

 

 

Extension plate of the transmitted-light base TL ST

Rear side of the transmitted-light base TL ST

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

1 Screws for changing the halogen lamp

 

 

 

 

 

 

 

 

 

 

 

2

Power connection socket

 

 

 

 

1

Adapter plate for easy assembly of focusing

 

3

Power switch

 

 

 

 

 

 

 

 

 

 

 

 

 

drives

 

 

 

 

 

 

 

 

2

Removable glass plate

 

 

 

 

 

 

 

 

3

Controller for light intensity

 

 

 

 

 

 

 

 

4

Adjustment for deflection mirror

 

 

 

 

 

 

 

 

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Leica TL ST Transmitted-light Base: Operation

Light intensity control

The left control adjusts the intensity of the 12 V/20 W halogen illumination.

1.Switch on the illumination of the base at the power switch.

2.Focus on the specimen.

3.Set the illumination to the desired intensity using the left control.

Transmitted-light control

The transmitted-light base TL BFDF has a slider that automatically moves the deflection mirror in the base when moved. The mirror is kept in the correct position at all times and permits smooth changeover between bright field and opaque transmitted light.

Bright field

Bright field is suitable for examining translucent objects featuring contrasting structures. The object is directly illuminated from below and is seen in its natural colors against a bright background.

ρρ Move the slider backwards until the desired effect is achieved.

Inclined transmitted light

Transmitted light that traverses the object obliquely will provide additional resolution and information when observing semitransparent, opaque objects.

ρρ Slowly pull the slider towards yourself until the desired effect is achieved.

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Leica TL ST Transmitted-light Base: Changing Bulbs

Changing the halogen lamp

Changing bulbs

Precautionary measures

 

 

Before you change the lamp, it is abso-

1. Unscrew the two screws on the heat sink

When inserting the lamp, ensure that

 

 

lutely necessary to unplug the power

and pull the heat sink out, along with the

the cables are inside the two metal

plug from the base to prevent the risk of electric­

lamp.

clamps. This prevents the cables from getting

shock!

 

caught during insertion.

 

 

The halogen lamp becomes very hot

 

 

 

 

 

 

during operation. Therefore, to avoid

 

 

 

 

 

 

 

 

 

 

being burned, let the base cool off for approx.

 

 

 

 

 

 

 

 

10 minutes after switching it off!

 

 

 

 

Do not touch new halogen lamps with your bare fingers—this drastically

reduces the service life of the lamp!

2.

Carefully pull out the lamp and mount by

 

pulling them upwards.

 

 

 

3.

Disconnect the lamp from the mount.

 

4.

Insert the new lamp into the mount and

 

 

reinsert the lamp holder.

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Leica TL BFDF Transmitted-light Base: Controls

1

2

3

1Adapter plate for easy assembly of focusing drives

2Standard stage 10 447 269

3Button to toggle between bright field and dark field

Extension plate of the Transmitted-light base TL BFDF

Button to toggle between bright field and dark field

Adapter at the focusing drive

Connector for cold light sources (light conductor active f = 10mm, end tube f = 13mm)

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Leica TL BFDF Transmitted-light Base: Operation

Light intensity control

Observe the user manual—in particular, all safety regulations—from the

manufacturers of the light guide and cold light source.

ρρ Switch on the cold light source according to the manufacturer's user manual and adjust the brightness.

Transmitted-light control

The Leica TL BFDF transmitted-light base has a control that switches the light from "bright field" to "dark field".

Bright field

Bright field is suitable for examining translucent objects featuring contrasting structures. The object is directly illuminated from below and is seen in its natural colors against a bright background.

ρρ Turn the control as far as it will go towards "BF" ("bright field").

Dark field

In dark-field illumination, a ring illuminator is used in such a way that the direct light does not reach the objective without a specimen. Only the structure of semitransparent, opaque objects disperses the light, making the object visible against a dark background.

ρρ Turn the control as far as it will go towards "DF" ("dark field").

Fingertip with bright field illumination

Identical subject with dark field illumination

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Leica TL RC™ / TL RCI™: Controls

 

 

 

 

 

 

 

 

 

1

 

2

 

 

 

 

 

1

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

2

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

3

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

4

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

6

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

3 4 5

 

 

6

 

 

7

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Vertical column - transmitted-light base

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

adapter plate

1

Power switch

 

 

 

1

Heat sink of the integrated halogen illumina-

2

Power connection socket

 

tion (only TL RCI™)

3

USB socket, type B

2

Extension plate for focusing drives

4

USB socket, type A

3

Standard stage 10 447 269

5

2 CAN bus

 

 

 

4

Filter holder

6

Screws for changing the halogen lamp

5Control of top and bottom flaps of the Rottermann Contrast™

6Button for mirror and horizontal movement of the mirror

7Transmitted-light base

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Leica TL RCI™: The Deflection Mirror

1

Inverted world?

Depending on the properties of the specimen (refractive index of the environment) and the perception of the viewer, it is possible that the switches described below for positive and inverted relief contrast are to be operated in reverse. This means that the lower switch, rather than the upper switch, controls inverted the relief contrast and vice-versa.

Tilted mirror

The built-in mirror features one flat and one concave side and can be rotated and moved. The concave side has been specially designed for the optical requirements of objectives with a high numerical aperture. The black rotary knob on the left side of the transmitted-light base can be used to rotate the built-in path-folding mirror and move it forwards/back.

The concave cutout on the handle indicates the concave side of the mirror, allowing intuitive operation at any time without eye contact.

The angle of light incidence in the specimen plane changes depending on the tilt and position of the mirror. As a result, switching between transmitted light bright field, oblique illumination and dark field-like illumination is possible.

Functions of the rotary knob

The rotary knob (1) fulfills the following tasks:

ρρ Turning the deflection mirror from the flat to the concave side

ρρ Slight tilting to guide the light beam through the specimen plane at a steeper or flatter angle

ρρ Moving the deflection mirror (forwards/ back)

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Leica TL RCI™: Color Intensity and Temperature

12

1Potentiometer for controlling the CCIC™ (Constant Color Intensity Control) illumination intensity

2Potentiometer for controlling the color temperature

The transmitted-light base TL RCI™ has two electronic potentiometers that control the color intensity (1) and color temperature (2).

The controller for the color temperature simultaneously serves as an electronic shutter:

ρρ

To interrupt your work, click the potentiom-

 

eter (2).

ρρ

Click again to switch on the illumination.

 

The electronics returns the color tempera-

 

ture to the previously configured settings.

Using a USB mouse (only TL RCI™)

The Leica USB mouse controls the CCIC™ and dimming function of the TL RCI™ base. Connect the mouse to the corresponding USB port of the base.

ρρ

The scroll wheel of the mouse is occupied

 

by default with the CCIC™ control system

 

and is used to control the illumination

 

intensity.

ρρ

To switch the illumination on or off, briefly

 

click the scroll wheel.

ρρ

To enter or exit dimming mode, click and

 

hold down the scroll wheel for longer than

 

2 seconds.

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Leica TL RC™ / TL RCI™: Operation

Setting the relief contrast

The two switches on the left side of the trans- mitted-light base TL RC™/TL RCI™ activate two built-in flaps. The outer switch (1) controls the inverted relief contrast, and the inner switch (2) adjusts the positive relief contrast.

Depending on the flap position, a part of the opening of the built-in Fresnel lenses are covered, which results in the different contrast effects. Phase structures typically act as spatial, relief-type images—in the positive relief contrast like hills, in the inverted relief contrast like valleys.

Increased contrast without relief is attained if both diaphragms are set to 45°. A gap-like illuminated area is created. By tilting the deflection mirror slightly, you can move the gap over the entire field of view and quickly toggle between positive and negative relief images. The dynamic effect makes it easy to distinguish phase structures from amplitude structures.

1

2

3

1Switch for adjusting the inverted relief contrast

2Switch for adjusting the positive relief contrast

3Deflection mirror

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Leica TL RCI™: Methods in Transmitted Light

Vertical bright-field illumination

Suitable for stained amplitude specimens with sufficient contrast.

The light beams are deflected vertically through the specimen. This results in an accurate bright field with maximum brightness.

Inclined transmitted light

Suitable for semitransparent, opaque specimens such as foraminifera and fish eggs. Move the deflection mirror until the desired data are visible.

Single-sided dark field

Suitable for fixed specimens and fine structures.

The flatter the angle at which the light beams are deflected into the specimen plane, the darker the substrate appears. A dark field-like transmitted light is created. Outlines, fine edges and structures are bright, in contrast with the dark background, through diffraction of the light beams on the dark background.

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Leica TL RCI™: Relief Images

Starting position

1.Push the deflection mirror all the way back towards the column.

2.Turn the deflection mirror into the notch position at an angle of 45°.

Positive relief contrast

Suitable for semitransparent and transparent specimens. The phase structures look like hills.

The effect can be strengthened or weakened by gently tilting the deflection mirror.

Negative relief contrast

Suitable for semitransparent and transparent specimens. These settings result in a negative relief contrast. Phase structures look like valleys.

The effect can be strengthened or weakened by gently tilting the deflection mirror.

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