Twophoton and Wide Field Microscope

Applications

• In vivo two-photon imaging
• Electrophysiological recording and imaging (culture, large in vivo preparations, etc.)
• Non-horizontal surface microscopy
• Simultaneous retinal stimulation and two photon microscopy
  (see  Euler et al., "Eyecup scope-optical recordings of light stimulus-evoked fluorescence signals in the retina", Pflügers Arch., 2008.)
• Whole animal imaging
• Immunology
• Embryology
• Patch-slice, fixed-stage microscopy
• Non-horizontal surface microscopy

Features

• Objective moves 22 mm in X, Y and Z
• Objective rotates about optical axis for imaging of non-horizontal surfaces and volumes
• Customizable open platform design
• Cambridge Technology XY scanners
• Two or four channel detector system with Hamamatsu PMTs and preamplifiers
• Sutter PS-2 / PS-2 LV dual channel PMT power supply
• National Instruments based data acquisition system

The Movable Objective Microscope (MOM) is a two-photon microscope capable of imaging deep within living specimens when combined with a Ti:Sapphire laser. The MOM design is unique in providing 3-dimensional objective movement and rotation allowing the specimen to remain stationary. Many highly regarded imaging laboratories around the world use MOM microscopes and we constantly work with our customers to adapt the design for their changing needs.

Starting in 2011, Sutter has begun offering the MOM Computer System with software (MCS). The MCS is designed to take advantage of the MOM. As a result, when combined with MCS, the MOM becomes a powerful tool capable of combining electrophysiology with large scale imaging experiments in both two and three dimensions.

The MOM Opto-mechanical Design

The MOM consists of two independent microscopes. The widefield half of the microscope consists of an Olympus vertical illuminator, Sutter Xenon arc lamp and camera mount to provide standard epifluorescence. The two-photon side of the microscope provides the optical pathway for guiding the excitation laser light from the table up into the scanning galvanometric mirrors and then expanding the beam through the scan lens and directed into the back of the objective. Following two-photon excitation, the emitted photons are directed by a dichroic mirror immediately above the objective into the detection pathway. The main body of the microscope translates on a rail system to allow easy access to the specimen prior to imaging.

The objective translates in X, Y and Z as well as rotates around the X axis. Two moving mirrors allow the microscope to maintain efficient delivery of the excitation light to the back aperture of the objective regardless of movement or orientation. The X, Y and Z movements used are the same as that in Sutters MP-285 micromanipulator so you know the movements are smooth, fine in scale, drift-free and highly reproducible. These movements permit Z-stacks and mosaic images of large regions of tissue to be recorded without the need for a moving stage.

The horizontal light path allows for rotation of the objective away from the standard vertical position. As a result of this rotation, the MOM can easily be converted from an upright to an inverted microscope and the objective positioned from 0 to 180 degrees. This positional freedom permits the imaging of nonhorizontal surfaces and volumes.

The MOM is both an inverted and upright microscope with 0 to 180° rotation

MOM Scanning Systems

During the last 10 years, scanning technology has changed dramatically: objectives changed requiring larger beam sizes and scanner technology advanced providing dependable resonant scanners. Unlike other two photon microscope designs, the MOM has lived through and adapted to changes in scanning technology. Throughout this development, Sutter has maintained two principles. First, existing scopes can be upgraded to new technology as it becomes available. Many original scopes with 3mm galvo scanners have been upgraded to either 6mm galvo scanners or resonant/galvo scanners. Second, Sutter continues to supply the original designs if they are needed for current research. We can provide a 3mm or 6mm conventional scanning MOM or a resonant/galvo scanning MOM all at competitive prices.

Imaging Software

Starting in 2011, Sutter began offering the MOM Computer System and Software (MCS). Before this software package was developed, most users relied on ScanImage or MPScope to generate scanned images. Customers valued the fact that the MOM would operate with open source freewares, however, there seemed to also be a market for a commercial package. Sutter MScan offered a number of functions that were not available in the existing freeware packages including photostimulation and the ability to combine imaging with electrophysiological recording and photostimulation. As resonant scanning became popular, and none of the freewares supported resonant scanning on the MOM, Sutter and MScan took the initiative. The latest version, MScan 2.0, coupled with a faster data acquisition system, allowed the MOM to generate fast, resonant images. To this day, the Sutter MOM with MScan2.0 remains the one platform available that can switch back and forth between conventional and resonant scanning.

The MOM® has always been compatible with ScanImage freeware, the two-photon imaging software developed by Karel Svoboda and collaborators. One of the reasons the MOM platform exists in its present form is the strong support from the ScanImage community. In 2014, Vidrio became the principle vehicle for support and new development of ScanImage. Sutter is happy to make Vidrio ScanImage Premium available to customers who wish premium support and the latest features. ScanImage freeware is also still available as SI5. Sutter provides packages that include the necessary data acquisition hardware to couple the MOM and other scanning microscopes to ScanImage Premium or SI5.

The MOM is available with four different detector path designs. The original 2 channel pentagon, which can become a four channel detector path. The Short path and the Wide path are two designs that increase the chances of capturing ballistic photons by getting the first collection lens either closer to the back aperture of the objective (Short path) or by using a larger aperture collection lens and dichroic (Wide path).

Sutter MOM packages include all of the equipment (less the Ti:Sapphire laser and objective) needed for a complete imaging system.

Cambridge Technology XY galvanometric and resonant scanners
(conventional with 3 or 6 mm mirrors or resonant with 5 mm mirrors)

Hamamatsu photomultiplier tubes (PMTs):
R6357 multialkali or H10770PA-40 (GaAsP) products
(Sutter is an authorized reseller for Hamamatsu)

Power supplies for PMTs:
Either a Sutter PS-2 (dual channel high-voltage power supply for R6357 PMTs) or
Sutter PS-2LV (dual channel low-voltage power supply for H10770PA-40 (GaAsP) PMTs) can be ordered

Hamamatsu, Sigmann or FEMTO pre-amplifiers, selection varies with software and type of scan

Data acquisition: National Instruments and Measurement Computing systems

TECHNICAL SPECIFICATIONS

Travel
22 mm on all three axes

Resolution
MP-285 controller
Low: 0.2 µm/step
High: 0.04 µm/step

MPC-200 controller
0.0625 µm/step

Maximum Speed
MP-285 controller
2.9 mm/sec

MPC-200 controller
5.0 mm/sec

Long Term Stability
1-2 µm/hour

Drive Mechanism
Precision worm gear capstan drive

Communication
MP-285: RS-232 Serial
MPC-200: USB

Electrical
115/230 V,  50/60 Hz power line

LIGHT SOURCES

LAMBDA LS    Lambda LS with 300 W  xenon arc lamp
     Lamp Life  1,000 h (500 h warranty), Longer life depends on application

LS-OF30     Lambda LS with 300 Watt ozone free lamp

HPX-L5     Lambda HPX-L5 High powered LED  light source

LB-421     Lambda 421 Optical Beam Combiner and controller

LB-OBC     Lambda OBC Optical Beam Combiner

FLED      Lambda FLED

FLED-DC   Dual Channel Lambda FLED

PS-2/PS-2LV PMT POWER SUPPLY
Electrical: 115/230 V, 50/60 Hz power line

MDR-3/MDR-6/MDR-R SCAN DRIVE CONTROLLER
Electrical:  115/230 V,  50/60 Hz power line

2 channel

4 channel

Short path (2 channel)

Ordering information

MOM BASIC SYSTEM FOR 2-PHOTON MICROSCOPY
Includes Moving Objective Microscope, 2 channel detector with PMTs, preamps and PS-2 Power supply, XY scanners with servo drive, wide field fluorescence unit
including vertical illuminator, Lambda LS 300 Watt Xenon Arc lamp, LLG and light guide adapter, C-mount for wide field camera, data acquisition system

MOM-3MM    MOM System with 3 mm XY scanners
MOM-6MM     MOM System with 6 mm XY scanners
MOM-RES-MCS     MOM System with Resonant scanners and MCS 2.0
MOM-RES-SIP     MOM System with Resonant scanners and ScanImage Premium

ACCESSORIES
MOM-SETUPKIT-M     Basic table optics for laser routing
MOM-ALIGNTOOL     MOM alignment tool (useful tool for aligning the laser in MOM scopes, especially those with resonant scanners).

Final pricing depends on detector path selected and does not include several devices necessary for a complete 2-photon microscope (i.e. Ti:Sapphire laser, objective, camera, trinocular head, table mount optics).

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