Responsable: Pablo González-Melendi de León - Profesor/a Titular de Universidad

pablo.melendi@upm.es 910679169 Ext:79169 / 910679107 Ext:79107 ( Lab Microscopía )

 

Personal:

González Carneros, Carlos - Técnico/a


Box mail of complaints, suggestions and congratulations


Charter of services


 
Service member of:

Spanish Network of Advanced Light Microscopy (REMOA)

Platform of Microscopy for Biosciences. Network of Laboratories and Infrastructures of the Community of Madrid

 

 

 

Leica TCS-SP8 confocal microscope

 

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Super-resolution confocal microscope Zeiss LSM 880 with fast Airyscan

 

Available equipment

 

Microtomes and sample preparation
  1. Microtome Leica HistoCore NANOCUT R
  2. Vibratome Leica VT1200 S
  3. Paraffin-wax dispenser Leica EG 1110
  4. Hybridisation oven Amersham
Microscopes: 
  1. Zeiss Axiophot vertical microscope [Images]
  2. Leica DM IRB inverted microscope
  3. Leica DM2000 vertical microscope
  4. Stereomicroscope Leica MZ10F
  5. Stereomicroscope Olympus SZX9
  6. Stereomicroscope Leica MZ9
Confocal microscope: 
  1. Spectral confocal microscope Leica TCS-SP8 [Images]
  2. Super-resolution confocal microscope Zeiss LSM 880 with fast Airyscan
CCD camera 
  1. CCD camera NightOwl LB 983 NC100. Berthold
Equipment for high sensitivity image acquisition (FLumaZone) 
  1. Stereomicroscope LEICA M205FA for fluorescence and bioluminiscence
  2. High sensitivity Electron Multiplying Hamamatsu ImagEM X2 EM-CCD camera
Fluorescence motorized microscope with a high sensitive camera 
  1. Inverted microscope Leica Dmi8
  2. Digital camera Hamamatsu ORCA Flash 4.0 LT

 

Click hereto download the Fiji software for confocal imaging management.

 


Microtome Leica HistoCore NANOCUT R
  1. Automatic or manual working mode.
  2. Ergonomic and independent control panel.
  3. Holders for paraffin-wax and resin embedded specimens.
  4. Holders for steel and diamond knifes.
  5. Minimum sectioning thickness 0,5 µm.
  6. Integrated stereomicroscope.

 

 

 

 

Vibratome Leica VT1200 S

  1. Thick sections from non-embedded specimens by blade (Guillete) vibration.
  2. Automatic or manual working mode.
  3. Ergonomic and independent control panel.
  4. Integrated stereomicroscope.

Paraffin-wax dispenser Leica EG 1110

Hybridisation oven Amersham

 

 

  • Fluorescence cubes:

Excitation light

Excitation filter

Dichroic mirror

Emission filter

UV

BP 365

395

LP 397

blue

450-490

510

LP 520

green

BP 546

580

LP 590

 

  • Objectives:

Type

Magnification/numerical aperture

Technique

Immersion

Plan-NEOFLUAR

2.5x/0.075

 

 

Plan-NEOFLUAR

5x/0.15

 

 

Plan-NEOFLUAR

10x/0.3

 

 

Plan-NEOFLUAR

20x/0.5

DIC*

 

Plan-NEOFLUAR

40x/0.75

DIC*

 

Plan-NEOFLUAR

40x/1.3

phase contrast

oil

Plan-NEOFLUAR

63x/1.25

 

oil

Plan-NEOFLUAR

100x/1.3

 

oil

 

*DIC = differential interference contrast

 

Coupled to colour CCD camera Leica DFC 300FX (1,4 Mpixels)

 

  • Fluorescence cubes:

Type

Excitation light

Excitation filter

Dichroic mirror

Emission filter

A

UV

BP 340-380

400

LP 425

I3

blue

BP 450-490

510

LP 515

N2.1

green

BP 515-560

580

LP 590

 

  • Objectives:

Type

Magnification/numerical aperture

Technique

Immersion

PL FLUOTAR

1.6x/0.05

 

 

C PLAN

4x/0.10

 

 

C PLAN

10x/0.22

phase contrast

 

N PLAN

20x/0.40

 

 

N PLAN

40x/0.55

phase contrast

 

N PLAN

100x/1.25

phase contrast

oil

 

  • Coupled to colour CMOS camera DC.20000i euromex (20 Mpixels).

 

  1. diascopic and episcopic illumination
  2. plan-apochromatic objective 1.0x/0.125
  3. zoom 0.8x – 8x
  4. fluorescence cubes

Type

Excitation light

Excitation filter

Dichroic mirror

UV

UV

360/40

420

GFP Plus

blue

460-500

510

GREEN

green

546/10

590

 
Coupled to colour CCD camera Leica DFC 400C (5 Mpixels).

 

  • Fluorescence cubes:

Type

Excitation light

Excitation filter

Dichroic mirror

Emission filter

A

UV

BP 340-380

400

LP 425

L5

blue

BP 480/40

505

BP 527/30

N2.1

green

BP 515-560

580

LP 590

 

  • Objectives:

Type

Magnification/numerical aperture

Technique

Immersion

C PLAN

10x/0.22

 

 

C PLAN

20x/0.4

phase contrast

 

C PLAN

40x/0.65

phase contrast

 

HI PLAN

63x/0.75

 

 

C PLAN

100x/1.25

 

oil

 
Coupled to colour CCD camera Leica DFC 300FX (1,4 Mpixels).
  1. episcopic illumination
  2. objectives 0.5x y 1.0x
  3. zoom 0.63x – 5.7x
  4. fluorescence cubes

 

Type

Excitation light

Excitation filter

Dichroic mirror

UV

UV

400-440

475

GFP

blue

460-490

510

  1. episcopic illumination
  2. objective PLAN 1.0x
  3. zoom 0.63x – 6x

 

Inverted microscope Leica DMI6000CS 

 

  1. touchscreen STP6000

  2. smart move (moves the stage in XY and Z) 

  3. metal halide bulb EL6000 (120 W)

  4. Fluorescence clubes:

Type

Excitation light

Excitation filter

Dichroic mirror

Emission filter

A4

UV

BP 360/40

400

BP 470/40

CFP

violet/blue

BP 436/20

455

BP 480/40

GFP

blue

BP 470/40

500

BP 525/50

I3

blue

BP 450-490

510

LP 515

N2.1

green

BP 515-560

580

LP 590

 

  • objectives:

Type

Magnification/numerical aperture

Technique*

Immersion

PL APO CS

10x/0.4

BF,FLUO

 

PL APO CS

20x/0.70

BF,FLUO

 

PL APO CS2

20X/0.75

BF,DIC,FLUO

water, glycerol, oil

PL APO CS2

40x/1.1

BF,DIC,FLUO

water

PL APO CS2

63x/1.4

BF,DIC,FLUO

oil

 

*BF = bright field, DIC = differential interference contrast, FLUO = fluorescence

 

Confocal module

 

FOV scanner: 
  1. speed up to 1.800 Hz (3.600 Hz in bidirectional mode)
  2. up to 7 fps @ 512 x 512 px
  3. image format up to 8.192 x 8.192 px
Lasers: 
  1. CW (405 nm). 50 mW
  2. Ar (458, 488 y 514 nm). 100 mW
  3. DPSS (561 nm). 20 mW
  4. He-Ne (633 nm). 10 mW
Detectors: 
  1. 4 spectral for fluorescence and reflection (2 PMT and 2 HyD)
  2. 1 (non-confocal) for transmitted light
LIAchroic beam splitter, which includes the dichroic mirrors: 
  1. substrate (405)
  2. 488/561/633
  3. 458/514/561
  4. 476/488
  5. RT 15/85 (reflection)
  6. 488/561
Galvanometric Z-stage: 
  1. min. step 20 nm
  2. range 1500 µm
Motorized XY scanning stage 
  1. speed: 10 mm/s
  2. resolution 0,02-0,04 µm
  3. reproducibility

Software 

LAS X with wizard for FRET & FRAP analysis and 3D viewer

 

Bioluminiscence, chemiluminiscence and fluorescence (GFP)
  1. resolution 1024 x 1024 px
  2. sensitivity 300 to 1050 nm
  3. quantum yield (QY) 90% @ 620 nm
  4. dynamic range 82 dB

 

 

 

 

 

 

 

 

 

 

  1. external control panel for motorized functions
  2. episcopic illumination by upright semirigid LED lamps
  3. transmitted light base with three types of illuminations: bright field, Rotermann contrast and dark field
  4. apochromat objective 1X: NA 0,17 y WD 61,5 mm
  5. apochromat objective 2X: NA 0,35 y WD 20,1 mm
  6. motorized zoom, magnification between 0,78X and 16X (20,5 range)
  7. motorized fluorescence route
  8. Fluorescence cubes:

 

Type

Excitation light

Excitation filter

Dichroic mirror

DAPI

UV

350/50

460/50

GFP

blue

470/40

525/50

mCherry

green

560/14

630/75

 

Features:

  • Electron Multiplying Back Thinned Transfer CCD technique
  • high frame rate format with 512 x 512 pixels of 16x16 μm
  • readout speed of the full image 70 fps
  • binning 1x1, 2x2 and 4x4
  • EM (electron multiplying) gain between 1 - 1200x
  • Quantum Yield (QY) > 90%
  • analog gain between 0,5x and 1x for EM mode and between 1x and 5x for non-EM mode
  • air (-65ºC) and water (-80ºC) cooling
  • 16 bits imaging

 

 

 

  1. Integrated touch screen to control all functions

  2. CoolLed pE300 ULTRA illumination

  3. Autofocus Leica AFC (Adaptive Focus Control)

  4. Fluorescence cubes:

     

Type

Dye

Excitation filter

Dichro

Emission filter

LED 405

DAPI, BFP

405/60

455

470/40

FITC/TXR

CFP, GFP

mCherry

455/50

560/40

495

595

520/40

635/65

 

  • Objectives:

Type

Magnification/N.A.

Immersion

Coverslip (mm)

Techniques *

HC PL Fluotar

5x/0.15

 

 

BF, Pol, DIC, FLUO

HC PL Fluotar

10x/0.32

 

0.17

BF, Pol, DIC, FLUO

HC PL Fluotar

20X/0.55

 

0.17

BF, Pol, DIC, FLUO

HC PL Fluotar

40x/0.80

 

0.17

BF, Pol, DIC, FLUO

HC PL APO

63X/1,40

oil

0.17

BF, FLUO


*BF = bright field, DIC = differential interference contrast, Pol = polarisation, FLUO = fluorescence

 

XY motorized stage:

  1. max. speed: 120 mm/s

  2. range: 130 x 85 mm

  3. resolution: 0,05 µm

Z motorized stage:
  1. step: 10 nm
  2. range: 12 mm
  3. four speeds: 2 coarse, 2 fine

Features:

  1. CMOS sensor FL-400
  2. effective number of pixels: 2048 x 2048
  3. effective area: 13,312 mm x 13,312 mm
  4. frame rate: 30 frames/s a máxima resolución
  5. quantum yield (QY): over 70% at 600 nm
  6. digital binning 2x2 / 4x4

 

  1. touch screen to control all functions

  2. metal halide lamp (120 W)

 

Fluorescence cubes:

Type

Excitation light

Excitation filter

Emission filter

DAPI

UV

356

BP 455/50

GFP

blue

BP 470/40

BP 525/50

DsRed

green

BP 530-585

LP 615

 

 
Objectives:

Type

Magnification/numerical aperture

Technique*

Inmersión

Plan-Apochromat

10x/0.45

BF,FLUO

 

Plan-Apochromat

25x/0.80

BF,DIC,FLUO

water, glycerol, oil

W N-Achroplan

40x/0.75

BF, FLUO

water

Plan-Apochromat

40X/1.2

BF,DIC,FLUO

water, glycerol, oil

Plan-Apochromat

63x/1.4

BF,DIC,FLUO

oil

 

*BF = bright field, DIC = differential interference contrast, FLUO = fluorescence

 

CCD camera Axiocam 503 colour
  1. pixels: 1936 x 1460
  2. pixel size: 4.54 μm x 4.54 μm
Confocal module

 

Lasers

  1. 405, 488, 458, 514, 561 and 633 nm

Detectors

  1. 3 spectral PMTs
  2. Airyscan detector for super-resolution
  3. 1 PMT for transmitted light

Motorized XY scanning stage

  1. speed: 100 mm/s
  2. resolution: 0,1 µm
  3. reproducibility

Motorized Z stage

  1. range: 500 μm
  2. speed: 5 mm/sec
  3. reproducibility: +/- 5 nm

Software

  1. ZEN black: image acquisition
  2. ZEN blue: image processing (3D, colocalisation)


Luminometer to perform screenings of immunofluorescence experiments (single or double labels), in order to select fluorescence-positive samples for subsequent observation and analysis on the confocal microscope.


Specifications:

 

  1. Fluorescence and Luminescence..
  2. FRET, TR-FRET and BRET.
  3. Excitation length range: 200-1000 nm
  4. Emission length range: 270-840 nm
  5. Microplates from 6 to 384 wells.
  6. Orbital shaking.
  7. Temperature control from 4 to 45°C.