Multiphoton and Deep Imaging

Multiphoton Excitation Objectives

Engineered to achieve optimum performance with multiphoton excitation imaging (MPE), A Line MPE objectives enable high-precision imaging of biological specimens to a depth of up to 8 mm for in vivo and transparent samples.

This example shows a Z–stack image of an in vivo mouse under anesthesia from the brain surface to the radiate layer of the hippocampus (CA1) Sample: Thy1–YFP H line 8 week old male Excitation wavelength: 960 nm	Deep Mouse Brain Imaging Deep in vivo, multiphoton brain imaging and optogenetics at high resolution require objectives that have high transmission of infrared (IR) light, a high numerical aperture (NA), and the ability to correct for the depth and scattering of tissue. This objective delivers ultra-broad IR transmission, enabling optogenetic stimulation with visible light down to 400 nm and IR imaging or stimulation beyond 1600 nm. The correction collar reduces the excitation volume, enabling stimulation of single cells or dendritic spines. Combined with the powerful and precise scanning capabilities of the FVMPE–RS microscope, the XLPLN25XWMP2 objective is the right tool for high-precision multiphoton imaging. Image data courtesy of Katsuya Ozawa and Hajime Hirase, Neuron–Glia Circuitry, RIKEN Brain Science Institute, Japan

Image data courtesy of Katsuya Ozawa and Hajime Hirase, Neuron–Glia Circuitry, RIKEN Brain Science Institute, Japan

This example shows a Z–stack image of an in vivo mouse under anesthesia from the brain surface to the radiate layer of the hippocampus (CA1)
Sample: Thy1–YFP H line 8 week old male Excitation wavelength: 960 nm

Deep Mouse Brain Imaging

Deep in vivo, multiphoton brain imaging and optogenetics at high resolution require objectives that have high transmission of infrared (IR) light, a high numerical aperture (NA), and the ability to correct for the depth and scattering of tissue. This objective delivers ultra-broad IR transmission, enabling optogenetic stimulation with visible light down to 400 nm and IR imaging or stimulation beyond 1600 nm. The correction collar reduces the excitation volume, enabling stimulation of single cells or dendritic spines. Combined with the powerful and precise scanning capabilities of the FVMPE–RS microscope, the XLPLN25XWMP2 objective is the right tool for high-precision multiphoton imaging.

Image data courtesy of Katsuya Ozawa and Hajime Hirase, Neuron–Glia Circuitry, RIKEN Brain Science Institute, Japan

Deep Observation of Fixed Transparent Specimens with Multiphoton Objectives to a Depth of 8 mm

Our multiphoton objectives help facilitate breakthrough research on brain function and other vital organs by enabling researchers to see as deep as 8 mm without slicing. The XLPLN25XSVMP2 and XLSLPLN25XSVMP2 objectives support many clearing reagents.

Whole Mouse Brain Imaging (XLPLN10XSVMP)

The objectives offer a wide field of view with 10X magnification, single-cell resolution with an NA of 1.0, and observations down to 8 mm. The objectives match a wide range of clearing reagent refractive indices (ne: 1.33 to 1.52).

Image data courtesy of Hiroshi Hama, Atsushi Miyawaki, RIKEN Brain Science Institute Laboratory for Cell Function Dynamics

	High-Resolution Deep Brain Imaging of a ScaleS-Treated Mouse Brain (XLSLPLN25XGMP) With a numerical aperture of 1.0 and an 8 mm working distance, the objectives enable deep, high-resolution imaging with a refractive index that matches that of many clearing reagents (ne: 1.41 to 1.52). A maximum intensity projection image (top). Six XY images at different Z positions (bottom). WM: white matter; GCL: granule cell layer, Hil: hilus, LHb: lateral habenular nucleus, MDC: mediodorsal thalamic nucleusImage data courtesy of Hiroshi Hama, Atsushi Miyawaki, RIKEN Brain Science Institute Laboratory for Cell Function Dynamics Reference: Nat Neurosci. 2015 Oct; 18 (10): 1518–29. doi: 10.1038/nn.4107. Epub 2015 Sep 14.

High-Resolution Deep Brain Imaging of a ScaleS-Treated Mouse Brain (XLSLPLN25XGMP)

With a numerical aperture of 1.0 and an 8 mm working distance, the objectives enable deep, high-resolution imaging with a refractive index that matches that of many clearing reagents (ne: 1.41 to 1.52).

A maximum intensity projection image (top). Six XY images at different Z positions (bottom). WM: white matter; GCL: granule cell layer, Hil: hilus, LHb: lateral habenular nucleus, MDC: mediodorsal thalamic nucleusImage data courtesy of Hiroshi Hama, Atsushi Miyawaki, RIKEN Brain Science Institute Laboratory for Cell Function Dynamics
Reference: Nat Neurosci. 2015 Oct; 18 (10): 1518–29. doi: 10.1038/nn.4107. Epub 2015 Sep 14.

A Line MPE Objectives Selection Guide

	Working Distance (mm)	Magnification	Objective Field Number*	Numerical Aperture	Immersion	Sample	Purpose
XLPLN10XSVMP	8	10X	18	0.60	Water to oil (ne: 1.33 to 1.52)	In vivo and cleared samples	Wide field of view observation
XLSLPLN25XGMP	8	25X	18	1.00	Silicone oil to oil (ne: 1.41 to 1.52)	Cleared samples	High–resolution observation
XLSLPLN25XSVMP2	8	25X	18	0.95	Water to silicone oil (ne: 1.33 to 1.41)	In vivo and cleared samples
XLPLN25XSVMP2	4	25X	18	1.00	Water to silicone oil (ne: 1.33 to 1.41)	In vivo and cleared samples
XLPLN25XWMP2	2	25X	18	1.05	Water	In vivo

*Maximum field number observable through eyepiece.

MPE Objectives for Inverted Microscopes

	Working Distance (mm)	Magnification	Objective Field Number^*	Numerical Aperture	Immersion	Applications
UPLSAPO30XSIR	0.8	30X	22	1.05	Silicone oil	MPE imaging in deep tissue with a wider field of view

*Maximum field number observable through eyepiece.

Maximize Resolution in Deep Imaging

Our TruResolution objectives maximize resolution and contrast for 3D imaging deep within thick specimens. The objectives are equipped with a motorized correction collar that can automatically and dynamically compensate for spherical aberration while maintaining the focus position.

Learn More About TruResolution Objectives

TruResolution Objectives
MPE Dedicated Objectives Equipped with an Automated Spherical Aberration Compensation Function

	Working Distance (mm)	Magnification	Objective Field Number*	Numerical Aperture	Immersion	Sample	Purpose
FV30AC10SV	8	10X	18	0.60	Water to oil (ne:1.33 to 1.52)	In vivo and cleared samples	Wide field of view observation
FV30AC25W	2	25X	18	1.05	Water	In vivo	High-resolution observation

*Maximum field number observable through eyepiece.

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FVMPE-RS

Maximized resolution and contrast with TruResolution objectives
High-speed imaging with a resonant scanner
Extended IR multiphoton excitation up to 1300 nm
Triple scanner option for multiphoton and visible light laser stimulation
High sensitivity with a 1600 coating on 25XMPE objectives and the scanning unit
4-axis auto-alignment of IR laser beams

Learn More

*Banner Image: 20 week old YFP–H mouse brain treated with ScaleS
By courtesy of Hiroshi Hama, Atsushi Miyawaki, RIKEN Brain Science Institute Laboratory for Cell Function Dynamics

Multiphoton Excitation Objectives

Deep Mouse Brain Imaging

Deep Observation of Fixed Transparent Specimens with Multiphoton Objectives to a Depth of 8 mm

Whole Mouse Brain Imaging (XLPLN10XSVMP)

High-Resolution Deep Brain Imaging of a ScaleS-Treated Mouse Brain (XLSLPLN25XGMP)

A Line MPE Objectives Selection Guide

MPE Objectives for Inverted Microscopes

Maximize Resolution in Deep Imaging

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TruResolution Objectives MPE Dedicated Objectives Equipped with an Automated Spherical Aberration Compensation Function

Related Products

FVMPE-RS

Redirecting

TruResolution Objectives
MPE Dedicated Objectives Equipped with an Automated Spherical Aberration Compensation Function