Leica TCS SPE Manual de usuario

MICROSYSTEMS

Published

by:

Leica

Microsystems

CMS

GmbH

Friedensplatz

D-68165

Mannheim

(Germany)

http://www.leica-microsystems.com

http://www.confocal-microscopy.com/

Responsible

for

contents:

Leica

Microsystems

CMS

GmbH

Leica

Microsystems

CMS

GmbH.

All

rights

reserved.

Copyrights

The

instructions

contained

the

following

documentation

reflect

state-of-the-art

technology

and

knowledge

standards.

have

compiled

the

texts

and

illustrations

accurately

possible.

Nevertheless,

liability

any

kind

may

assumed

for

the

accuracy

this

manual's

contents.

you

have

•5

any

comments

these

operating

instructions

any

our

other

documentation

would

pleased

receive

them.

The

information

this

manual

subject

modification

any

time

and

without

notification.

All

rights

this

documentation

are

held

Leica

Microsystems

CMS

GmbH.

Adaptation,

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text

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(in

whole

part)

print,

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(including

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Leica

Microsystems

CMS

GmbH.

Programs

such

LAS

and

LAS

are

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laws.

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rights

reserved.

Reproduction,

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translation

these

programs

prohibited

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prior

written

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Leica

Microsystems

CMS

GmbH.

■■9

The

term

"Windows"

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used

the

following

text

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trademark

the

Microsoft

Corporation.

Otherwise,

inference

with

regard

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free

usability

product

ñames

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ñames

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ñames

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trademarks

the

respective

manufacturers.

Leica

Microsystems

CMS

GmbH

The

product

¡s

protected

under

the

following

patents:

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6,222,961;

6,285,019;

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6,444,971;

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Further

patents

are

pending.

Contents

Important

information

about

the

operating

instructions

1.1

Intended

use

The

Leica

TCS

SPE

system

Safety

instructions

and

their

meaning

Safety

instructions

)

4.1

General

safety

¡nstructions

4.2

Láser

safety

4.3

Electrical

safety

4.3.1

General

specifications

4.4

Standards

4.5

Instructions

for

handling

glycerin solution

Láser

safety

5.1

Láser

and

láser

class

5.2

Safety

labels

the

Leica

TCS

SPE

system

5.2.1

Microscope

DMI4000

5.2.2

Microscope

DM2500

5.2.3

Microscope

DM5500

5.2.4

Sean

head

5.2.5

Supply

unit

5.2.6

Mirror

housing

Safety

devices

the

microscope

6.1

Transmitted

light

lamp

housing

the

inverted

microscope 37

6.2

Transmitted

light

lamp

housing

the

upright

microscope

6.3

Mirror

housing

the

upright

microscope

6-4

Safety

beam

guide

and

beam

collector

6.5

Láser

keyswitch

and

láser

emission

display

6.6

Safety

switches

6.7

Interlock

6.8

Remote

interlock

Work

safety

7.1.1

Specimen

área

7.1.2

Changing

the

specimen

7.1.3

Changing

theobjective

7.1.4

Changing

filter

cube/beam

splitter

7.2

Disconnecting

the

power

supply

System

overview

8.1

System

components

and

peripheral

devices

8.1.1

Microscope

8.1.2

Supply

unit

with

workstation

integrated

cft

8.1.2.1

Dongle

"'".!

8.1.3

Scanhead

Installation

location

9.1

Ambient

conditions

9.1.1

Air-conditioning

the

installation

location

9.1.2

Power

supply

9.2

Installation

the

system

Starting

10.1

Switching

the

system 65

10.2

Switching

off

the

system

LAS

11.1

Calling

the

online

help

11.2

Full-text

with

logically

connected

terms

11.3

General

structure

the

user

interface

Introduction

confocal

work

12.1

Preparation

12.1.1

Theobjective

12.1.2

Conventional

microscopy

12.1.3

Why

sean?

".".".'"I"

12.1.4

How

optical

section

created?

•

12.2

Acquiring

optical

sections

12.2.1

Data

acquisition

12.2.2

Illumination '"'

12.2.3

Beam

splitting

12.2.4

Emission

bands

12.2.5

The

pinhole

and

its

effeets

12.2.6

Image

detail

and

ráster

settings

12.2.7

Signal

and

noise

12.2.8

Profilecuts

"'.".'.".".'"".".'.'

12.3

ultipara

meter

fluorescence

12.3.1

Illumination

\ZZZZZ'".'.'.".

12.3.2

Emission

bands

(«

12.3.3

Sequential

capture

12.4

series

""".'.".'.".'"'".'.'."

ít

12.4.1

z-stack

12.4.2

About

section

thickness

12.4.3

...and

spacing

12.4.4

Data

volumes

12.4.5

Depictions

12.4.5.1

Gallery

12.4.5.2

Movie

12.4.5.3

Orthogonal

projections

12.5

Time

series

100

12.5.1

Sean

speed

100

12.5.2

Unes

100

12.5.3

Planes

100

12.5.4

Spaces

(time-space)

100

12.6

Spectral

series

100

12.6.1

Data

acquisition

and

utilization

101

12.7

Combinatorial

analysis

101

Care

and

maintenance

103

13.1

Filter

104

13.1.1

Changing

the

filter

104

13.2

Moisture

105

13.3

Dust

cover

105

13.4

Clinging

dirt

106

13.5

Cleaning

the

specimen

stage

106

13.6

Cleaning

glass

surfaces

106

13.7

Cleaning

the

objective

107

13.8

Removing

immersion

oil

107

j»

13.9

Handling

acids

and

bases

107

13.10

Cleaning

the

supply

unit

108

13.11

Cleaning

the

sean

head

108

13.12

Cleaning

the

monitor

108

13.13

Cleaning

the

keyboard

108

Disposal

109

Contact

índex

113

Important

information

about

the

operating

instructions

These

operating

instructions

are

essential

component

the

Leica

TCS SPE

system

and

must

read

carefully

before

the

microscope

¡s

put

into

operation

used.

The

emphasis

the

safety

instructions,

which

must

strictly

observed

during

all

work

with

the

Leica

TCS

SPE

system.

Keep

these

instructions

convenient,

safe

place.

Please

read

and

observe

the

instructions

provided

for

the

individual

components

and

peripheral

devices

and

keep

them

safe

place.

These

operating

¡nstructions

present

the

first

steps

for

activating

the

system

and

describe

the

basic

functions

the

Leica

Application

Suite

Advanced

Fluorescence

(LAS

AF)

software.

Detailed

descriptions

the

software

functions

have

not

been

included

these

instructions.

Instead,

please

refer

the

online

help

the

LAS

AF,

which

presents

the

most

up-to-date

explanations

and

instructions

the

corresponding

software

functions.

Please

read

Chapter

LAS

familiarize

yourself

with

the

structure

and

use

the

online

help.

1.1

Intended

use

The

system

was

designed

for

confocal

recording

(láser

scanning

images)

fluorescence-

marked

living

and

fixed

specimens

well

for

quantitative

measurements

the

área

life

science.

This

system

intended

solely

for

laboratory

use.

The

system

not

intended

for

use

for

vitro

diagnostics,

defined

the

Germán

medical

devices law

(Medizinproduktgesetz).

The

user

this

product

responsible

for

proper

and

safe

operation

and

safe

maintenance

the

system

well

for

following

all

applicable

safety

regulations.

The

user

fully

Hable

for

all

consequences

resulting

from

the

use

the

system

for

any

other

purposes

than

those

listed

the

Operating

Instructions

the

online

help.

The

manufacturer

assumes

responsibility

liability

for

any

use

outside

the

intended

use

outside

the

specifications

from

Leica

Microsystems

CMS

GmbH

any

risks

resulting

from

such

use.

such

cases,

the

Declaration

Conformity

shall

invalid.

Due

potential

hazards,

only

personnel

who

are

familiar

with

its

operation

and

who

nave

appropriate

training

should

assigned

responsibilities

with

the

Leica

TCS

SPE

system.

Such

training

can

obtained

from

the

manufacturer,

from

láser

protection

officer

from

recognized

outside

organizaron,

and

should

cover

least

the

following

points:

■

Familiarization

with

the operating

processes

the

system

(under

consideration

all

safety

instructions

provided

these

operating

instructions)

■

Correct

use

láser

protection

for

preventing

hazards,

use

warning

signs

etc.

■

Necessary

personal

protection

■

Biológica

effect

the

láser

eyes

and

skin

The

Leica

TCS

SPE

system

The

Leica

TCS

SPE

true

confocal

microscope

system

for

entry-level

áreas.

This

extremely

compact

system

particularly

easy

use

while

offering

all

the

advantages

confocal

microscope.

Ultrathin

z-sections

with

one-of-a-kind,

pinpoint

sharp

image

quality

are

recorded

conjunction

with

the

integrated,

high-precision

Galvo-z

stage

and

composed

into

three-

dimensional

visualizations

with

the

new

LAS

software.

The

system

achieves

máximum

signal

output

with

the

optimized

tuning

the

special,

highly

dynamic

photomultiplier,

which

comes

with

new

control

technology,

motorized

variable

pinhole

and

efficient

prisms.

The

spectral

detector

ensures

the

perfect

adjustment

the

emission

spectrum

today's

dyes

and

the

dyes

the

future.

The

TCS

SPE

can

equipped

with

four

lasers.

Long-lasting

solid-state

lasers

with

emission

wavelengths

488

nm,

532

and

635

are

included

with

the

standard

system.

optional

versión

with

additional

405

láser

also

available.

Compared

ó)

conventional

gas

lasers,

solid

state

lasers

provide

significantly

longer

service

life

and

excellent

stability.

Using

the

standard

AOTF,

possible

optimally

adjust

the

láser

intensity

infinitely

between

and 100

prevent

irradiation

and

unnecessary

bleaching.

Based

the

high-level

integration

the

supply

unit,

the

TCS

SPE

can

used

any

laboratory;

extensive

cooling

special

power

termináis

are

not

necessary.

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