Leica DM 6000B

Widefield Upright Microscope

 

Transmitted Light
Phase Contrast; Bright Field; Dark Field

 

Fluorescence Filters For
DAPI; CFP; GFP; FITC; YFP; CFP-YFP; CY3; TX2; mCherry

 

Advanced Techniques
Fluorescence recovery after photobleaching (FRAP)
Förster resonance energy transfer (FRET)

 

Training is required before use
An equipment guide is available in our Guides section
Contact BIC for training
System Booking
Location: 5.25A

 

 leica_dm_6000b.png

 

Warnings

  • Before you get started for the first time in this microscope consult the responsible people for proper training
  • ONLY turn ON the lamp if it has been off for more than one hour
  • Do not turn OFF the lamp if someone else will be using the microscope in the next few hours. To avoid unnecessary working hours or usage constraints, please contact the next user after using the microscope
  • Turn ON THE LAMP FIRST and then the remaining devices
  • THE LAMP must be the LAST TO TURN OFF
  • When closing the MetaMorph software, WAIT for the warning message and let the camera cooling down. In other words, close MetaMorph and DO NOT close any other window that will pop-up until the software closes by itself
  • Never LOOK DIRECTLY into the beam paths as they are capable of permanently damaging the human eye
  • DO NOT FORGET to clean the oil immersion objectives. Use ONLY the specific tissues (Kimtech®) for this purpose

 

Brief Description

A fully automated upright microscope equipped with motorized z-focus, coded 7x nosepiece, transmitted light axis and incident light axis.  The Leica DM6000 B provides the main transmitted light contrast methods and a user-friendly interface for fluorescence microscopy. Furthermore, the microscope is equipped with a laser unit emitting UV radiation which allows performing the FRAP and FRET techniques.

 

Suggestion for “Materials and Methods”

Images were acquired on a Leica DM 6000B upright microscope equipped with an Andor iXon 885 EMCCD camera and controlled with the MetaMorph V5.8 software, using the 100x 1.4 NA oil immersion objective plus a 1.6x optvar, the fluorescence filter sets GFP + TX2 and Contrast Phase optics.

 

Suggestion for “Acknowledgements”

This work was partially supported by PPBI - Portuguese Platform of BioImaging (PPBI-POCI-01-0145-FEDER-022122) co-funded by national funds from OE - "Orçamento de Estado" and by european funds from FEDER - "Fundo Europeu de Desenvolvimento Regional".

 

Filter Sets

Filter Cube

Excitation Filter

Dichromatic Mirror

Emission Filter

Excitation Colour

Emission Colour

A4; DAPI1;2

BP 360/40

400

BP 470/40

UV

Blue

CFP

BP 436/20

455

BP 480/40

Violet/Blue

Blue/Cyan

L5; GFP1; FITC1;2

BP 480/40

505

BP 527/30

Blue/Cyan

Green

YFP

BP 500/20

515

BP 535/30

Green

Green/Yellow

CFP-YFP3

BP 436/12 - 500/20

445 - 515

BP 467/37 - 545/45

Violet/Green - Green

Blue/Cyan – Green/Yellow

CY3; TRITC1

BP 545/30

565

BP 610/75

Green/Yellow

Orange/Red

TX2; FM 4-641; mCherry1

BP 560/40

595

BP 645/75

Green/Yellow

Red

1 Almost equivalent to this filter cube

2The filter cube may appear entitled to such name in spite of the less known corresponding filter set

3Filter set for the FRET technique

BP stands for a bandpass filter

 

Objectives

Magnification1;2

10x

40x

63x

100x

Leica System

HC

HCX

HCX

HCX

Class

PL APO (Apochromats)

PL Fluotar (Semi -Apochromats)

PL APO (Apochromats)

PL APO (Apochromats)

Aperture (A)

0.4

0.75

1.4

1.4

Immersion

Air

Air

Oil

Oil

Free Working Distance (mm)

2.2

0.4

0.1

0.09

Cover Glass (mm)

0.17

0.17

0.17

0.17

Contrasting Methods

Phase Contrast (PH1)

Phase Contrast (PH2)

Phase Contrast (PH3)

Phase Contrast (PH3)

Pixel size w/ 1.6x optovar (µm)3

 0.5 0.125 0.08 0.05

Pixel size w/ 1.25x optovar (µm)3

 0.64 0.16 0.1 0.064

Pixel size without  optovar (µm)3

 0.8 0.2 0.126 0.08

1Notice that the microscope is equipped with two Eyepieces: HC Plan S 10x/25 Br. M

2Notice that the microscope is equipped with two optvar options: 1.25x; 1.6x

3Pixel sizes were calculated for an Andor IXon EM 885 EMCCD camera

 

Camera

  • Brand: Andor Technology®
  • Model: iXon EM+ 885 EMCCD
  • Reference: DU 885 K-CSO-#VP
  • EMCCD Technology
  • 1004 x 1002 active pixels
  • 4 Frame Rate @ full resolution (frames per sec)
  • 8 x 8-pixel size (WxH; µm)