Instrument


Leica upright confocal microscope TCS SP8   

Room:1016、1017

Introduction

The laser confocal scanning microscopy (LCSM) is an optical imaging technique to increase optical resolution and contrast of a microscopy by means of using laser as excitation light, combining with a spatial pinhole in front of the detector to block out-of-focus light in image formation. Compared with the traditional fluorescence microscope, LCSM has better horizontal and axial resolution, contrast and imaging depth.

Application:

Both of Leica SP8 confocal microscope in the center for optical imaging are upright fluorescence microscope, which are suitable for observation of brain tissue sections, cell smears and so on, while they are not suitable for observation of cell culture. Equipped with multi-module stages and a 20x water objective lens with long working distance, making SP8 powerful to observe the tissues of anesthetized animals. In addition, SP8 can be used to observe transparent samples with a 25x objective.

Technical indicators:

Laser:405nm(50mW)、488nm(20mW)、552nm(20mW)、638nm(30mW),fast AOTF with the switching time<5ms.

Imaging model: transmitted light brightfield imaging and fluorescence imaging. The fluorescence imaging adopts prism splitting, and all detection channels can be selected during the wavelength of 400~800 nm freely with the spectral precision of 1 nm. Both the detection wavelength range and the center are continuously adjustable.

Filter Parameters: DAPI Ex: BP 360/40   Em: BP 470/40

                           GFP  Ex: BP 470/40    Em:BP 525/50

                           YFP  Ex:BP 500/20     Em: BP 535/30

                                 RHOD Ex:BP 546/10   Em: BP 585/40 

Detector: two PMTs (Photomultiplier detector) and two HyDs (Ultra-high sensitivity phosphorous gallium arsenide-avalanche diode hybrid GaAsP detector, Quantum detection efficiency reaches 45% at 500nm)

Objectives: 

5x (BABB, NA=0.15, DLS, WD=4.8mm) HC PL FLUOTAR;

10x (Air, NA=0.40, WD=2.74mm) HC PL APO CS2;

20x (Air, NA=0.75, WD=0.62mm) HC PL APO CS2;

L20x (Immersion, NA=0.95,WD=1.95mm) HCX APO IMM;

25x (Immersion, NA=1.0,WD=6mm,mot), CLARITY HC FLUOTAR IMM;

40x (Oil, NA=1.3, WD=0.24mm) HC PL APO CS2;


Resolution: the resolution of XY≥140nm, the resolution of Z reaches 350nm.

Scan resolution: 4×1~8192×8192. The scan resolution of all channels can reach 8192×8192 and 16 bits grayscale depth.

Scan mode: high resolution scan mode (Galvano-mirror) and high speed scan mode (Resonant-scanner). The speed of high resolution scan mode is 7fps@512×512 with the scan area 22mm. The speed of high speed scan mode is 28fps@512×512.

Scan zoom: the continuous range of scan zoom is 0.75x ~48x.

Image processing software: LAS X, including functions of 3D visualization, intracellular structure measurement,cross-color separation, full specimen navigation and stitches.

 


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