Leica M205 FA User Manual

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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