TriM Scope II 2-Photon Microscopy

LaVision BioTec GmbH, Astastraße 14, 33617 Bielefeld, Germany Fon: +49 / 521 / 91 51 39-0, Fax: +49 / 521 / 91 51 39-10 Email: info@lavisionbiotec.com, Internet: www.lavisionbiotec.com

Cloud Scanner Brighter images and 20 times less bleaching

The Cloud Scanner is a software controlled beam splitter that delivers 8 individual and simultaneously scanned beams for fast imaging. The foci pattern can be adapted to the application - it can be 8 foci in a single spot, in a line, or in an individual shaped cloud of foci. A delay between subsequent beams reduces photo damage. Scanning and data acquisition methods remain identical to a standard single beam laser scanning microscope that utilizes sensitive non-descanned PMTs - but the Cloud Scanner gains brightness. Since speed depends on brightness, the maximum frame rate can be increased substantially.

Targeted Path Scanning Package Multipoint timing

The Targeted Path Scanning Module measures the accurate angle of the xy galvo scanners and overlays the exact scan path and the image. Therefore, the laser focus can be precisely positioned for fast line imaging and laser treatment applications. In addition, it allows fast bidirectional scanning. Multipoint timing with: - up to 500 points - up to 2000 Hz - trigger out for patch clamp

Resonant Scanner Fast 2-photon imaging

The TriM Scope II scan head can be equipped with a resonant scanner that delivers more than 30 frames @ 512x512 pixel resolution. A combination of LaVision BioTec's patent pending Cloud Scanner technology and the resonant scanner delivers unmatched signal to noise ratio in high speed in vivo imaging applications.

Only a section of each pixel is covered with a conventional scanner

The Cloud Scanner beam array illuminates the entire pixel

EPI Ports are:PMT / TCSPC Port Ultrasensitive Port Ultrasensitive Port with swiveling objective mount

Detectors are:Standard Bialkali PMTs GaAsP PMTs GaAs PMTs GaAsP Hybride PMTs FLIM X16 TCSPC

Adapters are:2x, 3x, 4x, and 6x with fast mechanical shutter

Cloud Scanner

light source for transmitted light

Intravital Stage

TriM Scope II

CCD / sCMOS cameraSystem Diagram

Trinocular

Filter Wheel

EPI Ports Adapters Detectors

Transmission Port

The TriM Scope II is a modular 2-photon microscope platform that can be adapted to most applications. It comes with an upright or inverted stand - or both - and allows simultaneous scanning with up to two Ti:Sa lasers, one or two OPOs as well as various visible laser lines. To increase the imaging speed it can be equipped with LaVision BioTec's 64 Beam Splitter, LaVision BioTec's Cloud Scanner and a resonant scanner.

In addition to its imaging capabilities the TriM Scope II provides various line scanning and photo treatment modes that allow fast data acquisition, photo stimulation, uncaging and bleaching. Finally the TriM scope can be combined with a variety of detectors: · up to 2 imaging cameras for 64 beam mode, · up to 8 NDD PMT detectors, · up to 3 descanned PMT detectors, · and LaVision BioTec's FLIM X16 TCSPC detector.

FRAP, photoactivation, photostimulation, uncaging, and line scanning with Cloud Scanner

Fast intravital imaging with patent pending Cloud Scanner and resonant scanner

SHG and THG imaging with OPO and Ti:Sa laser

Integrated FLIM imaging with LaVision BioTec's FLIM X16 TCSPC detector

Ultrafast 64 beam imaging with up to 100 frames/s @ 2560x2160 pixel

OPO Package Deep NIR imaging

The first commercial OPO Package for NIR imaging (>1000 nm) has been implemented in 2005 by LaVision BioTec. It allows deep imaging and simultaneous excitation of blue/yellow/green and red dyes/fluorescent proteins. Custom-made optics ensure that OPO and Ti:Sa focus match exactly in the object plane.

FLIM Detector 78 MHz fast FLIM system

The FLIM Package is a fully integrated solution and implements LaVision BioTec's FLIM X16 TCSPC detector into the TriM Scope scanning microscope without limiting its imaging capabilities. During data acquisition the FLIM X16 detector is controlled by the standard ImSpector imaging software package and does not require any additional computer or software equipment. As the FLIM X16 detector allows 78 MHz count rate it does not compromise the scan rate.

Power Modulation Package FRAP and photo-treatment

The Power Modulation Package is designed to serve applications such as FRAP, photostimulation, photo-activation, and uncaging. Those applications require precise focus positioning and laser power adaptation in addition to the need to switch between laser treatment mode and imaging mode within milliseconds. The Power Modulation Package provides this functionality. It allows imaging or treating combinations of individual points, lines, and regions of interest in the field of view. In conjunction with AOMs the laser power can be adapted and switched between imaging and treatment within microseconds.

Fluorescence, second, and third harmonic imaging [courtesy of Peter Friedl]

LaVision BioTec intravital stage with conventional working distance and with 200 mm extended working distance

LaVision BioTec chimera system with one TriM Scope scan head serving an upright and an inverted microscope

64 Beam Option Fast in vivo imaging

The 64 Beam Option is the best choice for high frame rate applications like Ca2+ imaging in brain slices. LaVision BioTec's monolithic beam splitter divides the one laser beam into 64 individual beamlets that will be scanned simultaneously. This option requests a sCMOS CCD or EM-CCD camera as detector. Depending on the camera, the 64 beam option delivers up to 100 frames/s @ 2560x2160 pixel resolution.

Stands & Stages Up to 200 mm working distance

The TriM Scope II scan head can be combined with upright and inverted stands. LaVision BioTec provides its own upright Intravital Stand, Nikon FN1, Nikon Ti-U, Olympus BX 51WI, Zeiss Axio Examiner, and Zeiss Axio Observer. Most stands can be combined with LaVision BioTec's Intravital Stage or 3rd party stages for patch clamp applications.

Confocal Package 1-photon imaging

The TriM Scope can be equipped with up to 3 confocal detectors and LaVision BioTec's visible laser module that includes up to 5 different laser lines.

>> The modular multiphoton microscopy platform for functional and intravital imaging <<

TriM Scope II2-Photon Microscopy

OPO Package Deep NIR imaging

The first commercial OPO Package for NIR imaging (>1000 nm) has been implemented in 2005 by LaVision BioTec. It allows deep imaging and simultaneous excitation of blue/yellow/green and red dyes/fluorescent proteins. Custom-made optics ensure that OPO and Ti:Sa focus match exactly in the object plane.

FLIM Detector 78 MHz fast FLIM system

The FLIM Package is a fully integrated solution and implements LaVision BioTec's FLIM X16 TCSPC detector into the TriM Scope scanning microscope without limiting its imaging capabilities. During data acquisition the FLIM X16 detector is controlled by the standard ImSpector imaging software package and does not require any additional computer or software equipment. As the FLIM X16 detector allows 78 MHz count rate it does not compromise the scan rate.

software package and does not radditional computer or software As the FLIM X16 detector allows rate it does not compromise the

Power Modulation Package FRAP and photo-treatment

The Power Modulation Package is designed to serve applications such as FRAP, photostimulation, photo-activation, and uncaging. Those applications require precise focus positioning and laser power adaptation in addition to the need to switch between laser treatment mode and imaging mode within milliseconds. The Power Modulation Package provides this functionality. It allows imaging or treating combinations of individual points, lines, and regions of interest in the field of view. In conjunction with AOMs the laser power can be adapted and switched between imaging and treatment within microseconds.

Fluorescence, second, and third harmonic imaging [courtesy of Peter Friedl]

LaVision BioTec intravital stage with conventional working distance and with 200 mm extended working distance

LaVision BioTec chimera system with one TriM Scope scan head serving an upright and an inverted microscope

64 Beam Option Fast in vivo imaging

The 64 Beam Option is the best choice for high frame rate applications like Ca2+ imaging in brain slices. LaVision BioTec's monolithic beam splitter divides the one laser beam into 64 individual beamlets that will be scanned simultaneously. This option requests a sCMOS CCD or EM-CCD camera as detector. Depending on the camera, the 64 beam option delivers up to 100 frames/s @ 2560x2160 pixel resolution.

Stands & Stages Up to 200 mm working distance

The TriM Scope II scan head can be combined with upright and inverted stands. LaVision BioTec provides its own upright Intravital Stand, Nikon FN1, Nikon Ti-U, Olympus BX 51WI, Zeiss Axio Examiner, and Zeiss Axio Observer. Most stands can be combined with LaVision BioTec's Intravital Stage or 3rd party stages for patch clamp applications.

Confocal Package 1-photon imaging

The TriM Scope can be equipped with up to 3 confocal detectors and LaVision BioTec's visible laser module that includes up to 5 different laser lines.

Cloud Scanner Brighter images and 20 times less bleaching

The Cloud Scanner is a software controlled beam splitter that delivers 8 individual and simultaneously scanned beams for fast imaging. The foci pattern can be adapted to the application - it can be 8 foci in a single spot, in a line, or in an individual shaped cloud of foci. A delay between subsequent beams reduces photo damage. Scanning and data acquisition methods remain identical to a standard single beam laser scanning microscope that utilizes sensitive non-descanned PMTs - but the Cloud Scanner gains brightness. Since speed depends on brightness, the maximum frame rate can be increased substantially.

Targeted Path Scanning Package Multipoint timing

The Targeted Path Scanning Module measures the accurate angle of the xy galvo scanners and overlays the exact scan path and the image. Therefore, the laser focus can be precisely positioned for fast line imaging and laser treatment applications. In addition, it allows fast bidirectional scanning. Multipoint timing with: - up to 500 points - up to 2000 Hz - trigger out for patch clamp

Resonant Scanner Fast 2-photon imaging

The TriM Scope II scan head can be equipped with a resonant scanner that delivers more than 30 frames @ 512x512 pixel resolution. A combination of LaVision BioTec's patent pending Cloud Scanner technology and the resonant scanner delivers unmatched signal to noise ratio in high speed in vivo imaging applications.

Only a section of each pixel is covered with a conventional scanner

The Cloud Scanner beam array illuminates the entire pixel

LaVision BioTec GmbH, Astastraße 14, 33617 Bielefeld, Germany Fon: +49 / 521 / 91 51 39-0, Fax: +49 / 521 / 91 51 39-10 Email: info@lavisionbiotec.com, Internet: www.lavisionbiotec.com

EPI Ports are:PMT / TCSPC Port Ultrasensitive Port Ultrasensitive Port with swiveling objective mount

Detectors are:Standard Bialkali PMTs GaAsP PMTs GaAs PMTs GaAsP Hybride PMTs FLIM X16 TCSPC

Adapters are:2x, 3x, 4x, and 6x with fast mechanical shutter

Cloud Scanner

light source for transmitted light

Intravital Stage

TriM Scope II

CCD / sCMOS cameraSystem Diagram

Trinocular

Filter Wheel

EPI Ports Adapters Detectors

Transmission Port