Microscopy Core Facility

The Multi-User Microscopy Facility provides a broad spectrum of equipment for advanced microscopy studies and is open to all researchers at the Charité.

The Multi-User Microscopy Facility comprises an experimental setup for in  vivo experiments (two photon), a confocal setup for multicolor-imaging of fixated preparations and a normal fluorescence microscope.

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NeuroCure Multiuser Systems

Within the framework of NeuroCure, several shared microscope facilities are available for research use, and these setups are described below.

Multi-Photon in vivo set up

Optimized for in vivo preparations


Excitation
Two photon IR (690-1040nm)
Detection
Non-descanned-PMT green (480-580nm)
Non-descanned-PMT red (590-710nm)
Equipment
Upright microscope (Olympus BX 61)
10x and 40x water immersion objectives
L&N table and micromanipulators

 

Multi-Photon in vitro set up
Optimized for in vitro preparations

Excitation
Two photon IR (710-950nm) and
Two photon IR (720-950nm)
Detection
Non-descanned-PMT green (480-580nm)
Non-descanned-PMT red (590-710nm)
Equipment
Upright microscope (Olympus BX 61)
10x and 40x water immersion objectives
L&N table and micromanipulators
Confocal set up
Optimized for fixated preparation and cell culture

Excitation
Epifluorescence blue (450-490nnm)
Epifluorescence green (515-560nm)
Single-photon Ar (458, 476, 488,496, 514nm),
Solid state (568nm), HeNe (633nm)
Detection
3 PMT 400 - 800nm spectral descanned
Equipment
Invers microscope (Leica DMI 6000)
10x dry, 20x imm,  63x oil immersion objectives
Condensor dry NA 0.9
z-galvo table, mechanical xy-table (both Leica)
Gated STED Set-Up
High resolution Microscope optimized for fixated preparations

Excitation
Epifluorescence blue (450-490nm)
Epifluorescence green (515-560nm)
Single photon Ar (458, 476, 488, 496, 514nm)
Solid state (568nm), NeNe laser (633nm)
CW gated STED laser (592nm)
White light laser (450-650nm)
Detection
2 PMT 400-800nm spectral descanned
2 HYD (high sensitive) 400-800nm spectral descanned
Equipment
inverse microscope (Leica DMI 6000)
10x dry, 20x imm, 63x oil Immersion objectives
z galvo table, mechanical xy-table

Charité Multiuser Systems

The Multi-User Microscopy Facility provides a broad spectrum of equipment for advanced microscopy studies and is open to all researchers at the Charité.

The Multi-User Microscopy Facility comprises an experimental setup for in  vivo experiments (two photon), a confocal setup for multicolor-imaging of fixated preparations and a normal fluorescence microscope.

Multi-Photon set-up

Optimized for in vitro preparations and fixated preparations


Excitation
Epifluorescence blue (450-490nm)
Epifluorescence green (515-560nm)
Single-photon Ar (458, 476, 488, 514nm)
Two-Photon R (720-860nm)
Detection
2 PMT 400-700nm special descanned
non-descanned-PMT green (500-550nm)
non-descanned-PMT red (570-650nm)
Equipment
Upright microscope (Leica LFSA)
4x dry, 20x, 40x, 63x water immersion objectives
Confocal set-up

Optimized for fixated preparations


Excitation
Epifluorescence uv (340-380nm)
Epifluorescence blue (450-490nm)
Epifluorescence green (515-560nm)
Single-photon
Solid-state Laser (405, 488, 532, 635nm)
Detection
1 PMT 430 - 750nm spectral descanned
Equipment
Upright microscope (Leica DM 2500)
10x dry, 20x, 40x, 63x oil immersion objectives
Condensor dry NA 0.9
z-galvo table, mechanical xy-table (both Leica)
Fluorescence set-up

Optimized for fixated preparations


Excitation
Epifluorescence uv (360-370nm)
Epifluorescence blue (470-495nnm)
Epifluorescence green (540-550nm)
Detection
CCD Camera monochrome or color
Emission blue (420-460nm)
Emission green (510-550nm)
Emission red (575-625nm)
Equipment
Upright microscope (Olympus BX 61)
2.5x, 10x, 20x, 40x, dry 60x, 100x, oil immersion objectives
Neurolucida set-up

Optimized for fixated preparations

 
Excitation
Epifluorescence uv (360-370nm)
Epifluorescence blue (470-495nnm)
Epifluorescence green (540-550nm)
Detection
CCD Camera color
Emission blue (420-460nm)
Emission green (510-550nm)
Emission red (600-660nm)
Equipment
Upright microscope (Olympus BX61)
2.5x, 10x, 20x, 40x, dry 60x, 100x,
oil immersion objectives

General Considerations

Please find information regarding microscopy below.

Laser safety

The Tsunami (of Spectra-Physics) and Chameleon Ultra (of Coherent) lasers are 4 class high power lasers, whose beams can cause badly eye or skin damage.
In our facility the laser beam is completely enclosed and shielded up to the microscope objectives.


To prevent exposure to both direct and reflected beams please:

  • don’t enter the room, if there are warning notices at the door
  • use only what you are introduced in
  • consider the effects before each action
  • don’t take any covers from the equipment
  • don’t install or uninstall optical elements (objectives excepted)
  • don’t put mirrors or other reflective things under the microscope objectives
Software

Software to download

 

Please select the free software

LCS Lite Download

LASAF Lite Download

CellSens Free Download

 

Resolution
Notes on confocal resolution

Lateral resolution:

FWHM = 0.4 * λ / NA

Axial resolution:

FWHM = 0.45 * λ / n (1-cos α)

NA = n*sinα

FWHM:
full width half maximum (or spatial resolution)

NA:
numeral aperture of the chosen objective

n:
refraction index of the sample medium (for air: n = 1, for immersion oil: n = 1.5)

λ:
laser wavelength

The improvement in spatial resolution corresponds lateral to 1.4x and axial to 6x compaired with the conventional fluorescence microscopy. You can adjust the spatial resolution in the LCS software using “Zoom” and “Format”.
As a simple rule you can use:

Lateral: resolution/3 = optimal size of the voxel
Axial: resolution/3 = optimal choice of the z-scan

Please note: the spatial resolution depends on the used wavelength.