Bergamoイメージングシステムを使用することで、優れた試料イメージ(「イメージングギャラリ」タブ参照)を得ることができ、それらは数多くの出版物に掲載されています(「関連出版物」タブ参照)。 右下の「共同研究で生まれたイノベーション」の動画でご覧いただけるように、当社のライフサイエンス部門はUniversity College LondonのDr. Michael Hausserの研究グループと連携し、彼らのニーズに合ったカスタム仕様のBergamo顕微鏡システムを設計してきました。当社のBergamoイメージングシステムについてのご質問は、ボックス内のContact USボタンをクリックして当社までお問合せください。
1回のベッセルビームによる走査画像(左)は、ガウシアンビームによる45枚の光学画像の断片を積み重ねて得られた体積走査画像(右)と同じ構造情報を捉えており、これは全走査時間を45分の1に短縮できることを示しています。これらの画像は脳切片の300 μm x 300 μmの範囲を走査したものです。ガウシアンビームによる画像を積み重ねた画像の深さはスケールバーで示されています。試料ご提供:Qinrong Zhang, PhD and Matthew Jacobs; the Ji Lab, Department of Physics, University of California, Berkeley.
Bergamo IIシリーズ顕微鏡システムは、Ø25 mm蛍光フィルタならびに25 mm x 36 mmダイクロイックミラーが入っている業界標準の蛍光フィルターセットに対応します。当社の検出モジュールは他社設計とは異なり磁石固定式のホルダとなっているため、素早く簡単にフィルタを交換して異なる測定が行えます。
当社では広域用Ø32 mm蛍光フィルタならびに32 mm x 42 mmダイクロイックミラー用の検出モジュールもご用意しており、より大きい集光角度で多くの信号検出が可能です。
Bergamo IIシリーズ顕微鏡は、M34 x 1.0、M32 x 0.75、M25 x 0.75ならびにRMSネジ付きの無限遠補正対物レンズに対応します。これにより多光子顕微鏡に使用される多くの低倍率、高NA対物レンズを装置に取り付けることが可能になります。当社の走査用光学素子は視野数が20と大きく、これらの多光子イメージング用対物レンズの光学設計を利用することで、同じ対物レンズを使用した他社製顕微鏡と比べても集光性能が高くなっています。
1.2 mm In Vivo Deep Brain 3D Image Stack, Courtesy of Dr. Hajime Hirase, Katsuya Ozawa,RIKEN Brain Science Institute, Wako, Japan
Application Summary
Scientists investigate the structure of the brain to understand functions of neuronal proteins as well as the causes of neurological diseases. Due to the difficulty of imaging through brain tissue caused by light scattering, this study often requires a multi-modality configuration allowing for a range of experimental conditions using any combination of multiphoton, confocal, and epi-fluorescence imaging. The three example multiphoton microscope configurations in the table below are designed to accommodate the needs of Structural Biology. Each configuration features fast-Z power ramping to accomplish high-resolution imaging deep within a sample. Our Two- and Three-Photon Imaging configuration uses both galvo-resonant and galvo-galvo scanners and infrared wavelength scanning optics to image second- and third-harmonic generation (SHG and THG). Alternatively, our Dual-Path Multiphoton Microscope with Confocal Imaging is outfitted with a confocal path that accommodates up to 4 laser lines and a 4-channel PMT detection module. The addition of a six-cube epi-illuminator module and sCMOS Quanatlux camera allows this system to perform epi-fluorescence imaging. Our Simple XYZ Imaging configuration is well-balanced for both in vitro and fixed stage in vivo microscopy research. With a removable transmitted illumination module, this versatile system can support a wide variety of experimental techniques, imaging modalities, and sample subjects.
Stitched Confocal Fluorescence Image of Rat Retina Stained with DAPI, Alexa Fluor® 555 and Alexa Fluor® 633, Courtesy of Dr. Jennifer Kielczewski, National Eye Institute, National Institutes of Health, Bethesda, MD
Application Summary
Researching neurological disorders involves measuring neural function using two-photon calcium imaging. This research requires fast image acquisition and photostimulation. We recommend three of our multiphoton microscope configurations for this application (see the table below). Each configuration offers a large working space and rotating microscope body, making it ideal for in vivo animal studies. Our Multi-Target Photoactivation configuration features a spatial light modulator (SLM), which allows the activation of groups of neurons at varying depths within a single field of view. For increased penetration of samples with scattering tissues, the three-photon capability of our Two- and Three-Photon Imaging configuration is recommended. Our Random-Access Scanning configuration uses a resonant-galvo-galvo scanner to take multiple high-resolution images within a single field of view. This scanner provides all the speed of a resonant-galvo scanner, while enabling multiple user-defined fluorescence activation regions for correlating neural responses in multiple regions of the brain.
Two-Photon Image of Neurons Expressing Thy1-YFP in a Cleared Region of the Hippocampus, Courtesy of the 2017 Imaging Structure and Function in the Nervous System Course at Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Application Summary
Scientists interested in this application focus on tracing neuronal pathways and researching dendritic spine plasticity. Imaging systems used in this type of research are capable of two-photon calcium imaging and/or confocal fluorescence imaging. High-resolution and high-sensitivity are crucial features for these systems. We offer three multiphoton configurations that are ideal for this application (see the table below). Each configuration may be used in photon-limited environments as they feature sensitive detection modules, in addition to our 10° or 14° full field-of-view collection optics with ultrasensitive GaAsP PMTs. Our Dual-Path with Confocal Imaging configuration allows for a range of experimental conditions to be observed using any combination of multiphoton, confocal, and epi-fluorescence imaging, along with photoactivation. Alternatively, the Video and High-Speed Imaging and Simple Z-Axis Imaging configurations provide a small footprint and large throat depth for in vivo two-photon imaging.
Laser-Scanned Two-Photon SHG+Dodt Gradient Contrast of Zebrafish Embryo
Application Summary
Neurogenetic studies require a wide variety of experimental setups for in vivo research. Multiphoton imaging is ideal for studying live organisms, especially embryos, as it reduces the occurrence of photobleaching and phototoxicity that is common with other light microscopy techniques. There are three multiphoton configurations we suggest for Neurogenetic applications (see the table below). The small footprint and large throat depth of each system provide ample room for sample mounts and experimental apparatus, such as the large setup used for Drosophilia studies by Chen et al. (click here for supplementary videos). Additionally, these systems provide video-rate, sequential two-photon imaging to study fast dynamic biological and chemical processes in vivo without damaging the sample.
Imaging Visually Evoked Synaptic Calcium Transient In Vivo, Using Dendrite Labels with GCAMP6, 200 µm Deep in Layer 2/3 Visual Cortex, Courtesy of David Fitzpatrick, Max Plank Institute for Neurobiology, Jupiter, FL, USA
Application Summary
In vivo functional imaging of organisms and the individual neurons linked to specific organism behaviors requires high-speed and high-sensitivity imaging. We offer six configurations for this application (see the table below). These configurations are equipped with an 8 or 12 kHz galvo-resonant imaging scanner and our 10° or 14° wide-angle collection optics to enable fast, high-resolution imaging. Each microscope system features a large throat depth, 5” of vertical travel, and smooth movement along the Z-axis to create a large working space ideal for in vivo volume imaging deep into highly scattering samples of neural tissue. For an improved range of movement around a sample, we recommend a configuration with a rotating body.
Simultaneous Photostimulation of 100 Cells Co-Expressing GCaMP6f (Green) and C1V1 (Red), Courtesy of Lloyd Russell, Dr. Adam Packer, and Professor Michael Häusser, University College London, United Kingdom
Application Summary
By studying synapses and circuits, researchers are able to understand neuronal activity. Imaging synapses and circuits often requires simultaneous stimulation of populations of neurons. To achieve this, we offer three configurations of our multiphoton microscope capable of fast image acquisition of multiple regions within a single field of view (see table below). Our Multi-Target Photoactivation configuration features a spatial light modulator (SLM), which allows multiple sites in a sample to be photoexcited simultaneously. With the SLM, each beamlet can be shaped to improve the efficacy of photoactivation, a crucial feature for activating neural populations at varying depths within a single field of view (FOV). Our High-Speed, Random-Access Scanning configuration uses a resonant-galvo-galvo scanner to take multiple high-resolution images within a single field of view. This scanner provides all the speed of a resonant-galvo scanner, while enabling multiple user-defined photoactivation regions. Lastly, our In Vivo Two-Photon Imaging configuration uses a galvo-resonant scanner for high-speed imaging.
Cochlear Organotypic Culture Loaded with Fluo4-AM and DM-Nitrophen AM, Calcium Release Triggered Using Galvo-Galvo Uncaging Pathway in Outer Hair Cell Indicated by Red Box After ~13 Seconds, Courtesy of Federico Ceriani and Walter Marcotti, University of Sheffield
Application Summary
In this application, scientists are interested in neural connections and intercellular movement. With multiphoton imaging, they are able to trace the direction and speed of ions moving through channel membrane proteins or neurotransmitters moving from one neuron to another. This research requires a microscopy system that has high-resolution and high-speed imaging of multiple fields of view (FOVs). We recommend six of our multiphoton configurations for this application (see the table below). This area of research often requires both in vivo and in vitro imaging within the same study, so within each configuration, our transmitted light modules can be installed or removed by the user in just a few minutes, making it exceptionally easy to switch between the two imaging modalities. These configurations are capable of high-frame-rate imaging and targeted laser activation for photostimulation, making them ideal for correlating neural responses in multiple regions of the brain.
Top: Astrocytes are labelled with SR101 (red). Arrows point to astrocytes that had Ca2+ elevations during tDCS. The numbers correspond to the cells and neurogliopil regions plotted in the graphs below. Bottom: Fluorescent intensity (ΔF/F) traces of astrocytes (orange), neurons (green) and neurogliopil (brown). Figure Courtesy of Monai H. et al. (See Below)
Application Summary
The study of cell biology, muscles, and glia often involves imaging in vitro or in vivo samples tagged with multiple fluorophores. Through multiphoton imaging with these fluorescent markers, researchers are able to observe gene expression related to neural function in different areas of the brain. We recommend two of our configurations for this application, see the table below. With two to four channel detection modules and fast sequential imaging using our Tiberius® Tunable fs laser, both of these configurations offer the flexibility necessary for experiments in this field. In addition, each configuration has a small footprint and a large throat depth to provide ample room for numerous sample mounting options, including in vivo imaging of mammalian brains via transcranial windows.
Drug discovery research is rapidly expanding and often requires a wide variety of experimental setups and imaging techniques. Two-photon imaging is frequently used to measure the characteristics of drug applications, including depth of drug penetration and the area of its spread throughout the cortex. We offer two configurations that are suitable for this application, see the table below. These configurations are are well-balanced for both in vitro and fixed stage in vivo microscopy research. The modularity of the Bergamo systems' removable trans-illumination module provides versatility in regard to experimental techniques, imaging modalities, and sample subjects. The Gibraltar breadboard platform line is ideal for mounting samples and supplementary equipment within these configurations.
ベッセルビームによるボリュームイメージング技術を用いた神経生物学でのin vivo 研究(オプション)
デュアル光路ランダムアクセス走査構成の仕様詳細
Configuration Specifications
Highlighted System Specifications
FOV
20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed
Resonant-Galvo-Galvo Scanner: 8 kHz: 30 fps at 512 x 512 Pixels or 12 kHz: 45 fps at 512 x 512 Pixels Galvo-Galvo Scanner: 3 fps at 512 x 512 Pixels
Scan Resolution
Bi-Directional: 8 kHz RGG and GG: Up to 2048 x 2048 Pixels 12 kHz RGG: Up to 1168 x 1168 Pixels Uni-Directional: 8 kHz RGG and GG: Up to 4096 x 4096 Pixels 12 kHz RGG: Up to 2336 x 2336 Pixels
Photoactivation
Targeted: Simultaneous IR, Sequential Visible Full Field: During Scanner Flyback
20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed
Galvo-Resonant Scanner: 8 kHz: 30 fps at 512 x 512 Pixels or 12 kHz: 45 fps at 512 x 512 Pixels Galvo-Galvo Scanner: 3 fps at 512 x 512 Pixels
Scan Resolution
Bi-Directional: 8 kHz Galvo-Resonant and Galvo-Galvo: Up to 2048 x 2048 Pixels 12 kHz Galvo-Resonant: Up to 1168 x 1168 Pixels Uni-Directional: 8 kHz Galvo-Resonant and Galvo-Galvo: Up to 4096 x 4096 Pixels 12 kHz Galvo-Resonant: Up to 2336 x 2336 Pixels
Photoactivation
Targeted: Simultaneous IR, Sequential Visible Full Field: During Scanner Flyback
Microscope Components
Microscope Body
XYZ Motion
Primary Scan Path (Multiphoton)
Galvo-Resonant (8 or 12 kHz)
Secondary Scan Path
Galvo-Galvo (Ø4 mm or Ø5 mm)
Scan Path Wavelength Range
450 - 1100 nm or 680 - 1600 nm on Primary and/or Secondary Path
Pockels Cell
Slow (250 kHz) (High-Speed and Wide-Wavelength-Range Options Available)
Variable Attenuator
Motorized
Multiphoton Detection
2 Non-Cooled GaAsP PMTs
Confocal Detection
4-Channel Multialkali PMTs with Variable-Size Pinhole Wheel
Collection Optics Module
14°
Objective Holder
Single
Objective
Nikon 25X (with Piezo Objective Scanner)
Sample Stage
Rigid Stand with Breadboard Insert
Widefield Imaging
Six-Filter-Turret Epi-Illuminator Module Broadband Light Source Quantalux™ sCMOS Scientific Camera
20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed
8 kHz Scanner: 30 fps at 512 x 512 Pixels or 12 kHz Scanner: 45 fps at 512 x 512 Pixels
Imaging Resolution
Bi-Directional: 8 kHz Scanner: Up to 2048 x 2048 Pixels 12 kHz Scanner: Up to 1168 x 1168 Pixels Uni-Directional: 8 kHz Scanner: Up to 4096 x 4096 Pixels 12 kHz Scanner: Up to 2336 x 2336 Pixels
Photoactivation
Full Field: 4-Channel
Microscope Components
Microscope Body
XYZ Motion
Imaging Scanners
Galvo-Resonant (8 or 12 kHz)
Scan Path Wavelength Range
450 - 1100 nm
Pockels Cell
Slow (250 kHz) (High-Speed and Wide-Wavelength-Range Options Available)
Transmitted Light Module with Laser Scanning Dodt Single-Cube Epi-Illuminator Module Four-Channel LED Source Quantalux™ sCMOS Scientific Camera
Laser
Tiberius® fs Tunable Laser
こちらの構成は in vitro ならびに固定式ステージのin vivo顕微鏡観察のどちらにも適しています。脱着式の透過光モジュール付きのシステムは、さまざまな実験手法、イメージング手法、そして対象物に対応する汎用性があります。
Click to Enlarge 顕微鏡下の広い空間により、補助機器を使用した大型のセットアップを対物レンズ下に設置しやすくなります。こちらでは、ショウジョウバエ用のVRシアターステージと2つの補助カメラを取り付けています。セットアップは、ThorSync(ThorImageLS)ソフトウェアのアドオン)を使用した画像取得機能と同期可能です。
20 mm Diagonal Square (Max) at the Intermediate Image Plane
Imaging Speed
8 kHz: 30 fps at 512 x 512 Pixels or 12 kHz: 45 fps at 512 x 512 Pixels
Imaging Resolution
Bi-Directional: 8 kHz Scanner: Up to 2048 x 2048 Pixels 12 kHz Scanner: Up to 1168 x 1168 Pixels Uni-Directional: 8 kHz Scanner: Up to 4096 x 4096 Pixels 12 kHz Scanner: Up to 2336 x 2336 Pixels
Photoactivation
Full Field: 4-Channel
Microscope Components
Microscope Body
Z-Axis Motion
Imaging Scanners
Galvo-Resonant (8 kHz or 12 kHz)
Scan Path Wavelength Range
450 - 1100 nm
Pockels Cell
Slow (250 kHz)
Variable Attenuator
Motorized
Detection
2 Multialkali PMTs (GaAsP PMTs and Cooled PMTs Available)
Collection Optics Module
8° with Shutter
Objective Holder
Single
Objective
Olympus 20X
Sample Stage
Rigid Stand Slide Holder with XY Translation Stage
Widefield Imaging
Single-Cube Epi-Illuminator Module Four-Channel LED Source Quantalux™ sCMOS Scientific Camera
Marvin JS, Scholl B, Wilson DE, Podgorski K, Kazemipour A, Müller JA, Schoch S, Quiroz FJU, Rebola N, Bao H, Little JP, Tkachuk AN, Cai E, Hantman AW, Wang SSH, DePiero VJ, Borghuis BG, Chapman ER, Dietrich D, DiGregorio DA, Fitzpatrick D, and Looger LL. "Stability, affinity, and chromatic variants of the glutamate sensor iGluSnFR." Nat Methods. 2018 Oct 30; 15: 936–939.
Moeyaert B, Holt G, Madangopal R, Perez-Alvarez A, Fearey BC, Trojanowski NF, Ledderose J, Zolnik TA, Das A, Patel D, Brown TA, Sachdev RNS, Eickholt BJ, Larkum ME, Turrigiano GG, Dana H, Gee CE, Oertner TG, Hope BT, and Schreiter ER. "Improved methods for marking active neuron populations." Nat Commun. 2018 Oct 25; 9: 1–12.
お持ちの顕微鏡に対応する最新のThorImageLSについては当社までお問い合わせください。ThorImageLS 4.0は、バージョン3. x 、2.xならびに1.xに新規機能を大幅に追加しており、旧モデルの顕微鏡に対応できない場合があります。当社では旧モデルをお持ちのお客様のために旧バージョンのソフトウェアのサポートを継続しております。
New Hardware Support
Added Support for Windows® 10 OS
Added Support for CS895MU and CS505MUMonochrome Cameras (Requires ThorCAM 3.2)
Allows for Hot Pixel Correction
Added Support for CSN210 Motorized Dual-Objective Nosepiece
Resonant-Galvo-Galvo Scanner, Galvo-Resonant Scanners, Galvo-Galvo Scanners, or Spatial Light Modulator; Single or Dual Scan Paths
Scan Speed
8 kHz Resonant-Galvo-Galvo or Galvo-Resonant
2 fps at 4096 x 4096 Pixels 30 fps at 512 x 512 Pixels 400 fps at 512 x 32 Pixels
12 kHz Resonant-Galvo-Galvo or Galvo-Resonant
45 fps at 512 x 512 Pixels 600 fps at 512 x 32 Pixels
Galvo-Galvo
3 fps at 512 x 512 Pixels 48 fps at 512 x 32 Pixels 70 fps at 32 x 32 Pixels Pixel Dwell Time: 0.4 to 20 µs
Galvo-Galvo Scan Modes
Imaging: Line, Polyline, Square, or Rectangle Non-Imaging: Circle, Ellipse, Polygon, or Point
Field of View
20 mm Diagonal Square (Max) at the Intermediate Image Plane [12 mm Diagonal Square (Max) for 12 kHz Scanner]
Scan Zoom
1X to 16X (Continuously Variable)
Scan Resolution
Up to 2048 x 2048 Pixels (Bi-Directional) [Up to 1168 x 1168 Pixels for 12 kHz Scanners] Up to 4096 x 4096 Pixels (Unidirectional) [Up to 2336 x 2336 Pixels for 12 kHz Scanners]
Compatible Objective Threadings
M34 x 1.0, M32 x 0.75, M25 x 0.75, and RMS
Multiphoton Signal Detection
Epi-Detection
Up to Four Ultrasensitive GaAsP PMTs, Cooled or Non-Cooled
Forward Direction
Two Ultrasensitive GaAsP PMTs
Maximum of Four PMTs Controlled by the Software at a Given Time
Collection Optics
8°, 10°, or 14° Collection Angle (Angles Quoted When Using an Objective with a 20 mm Entrance Pupil) Easy-to-Exchange Emission Filters and Dichroic Mirrors
Confocal Imaging
Motorized Pinhole Wheel with 16 Round Pinholes from Ø25 µm to Ø2 mm Two to Four Laser Lines (488 nm Standard; Other Options Range from 405 nm to 660 nm) Standard Multialkali or High-Sensitivity GaAsP PMTs Easy-to-Exchange Emission Filters and Dichroic Mirrors
Widefield Viewing
Manual or Motorized Switching Between Scanning and Widefield Modes Illumination Provided via LED or Liquid Light Guide C-Mount Threads for Scientific Cameras
Transmitted Light Imaging
Differential Interference Contrast (DIC) or Dodt Gradient Contrast Widefield or Laser Scanned Illumination Provided by Visible and/or NIR LEDs Compatible with Air or Oil Immersion Condensers
Three-Photon Imaging
Scan Optics for 900 - 1900 nm Range Achieve Reduced Background Scatter for Greater Sensitivity in Deep Tissue Imaging
Volume Imaging Using Bessel Beams
3D Volumetric Functional Imaging at Video Frame Rates Enhanced Temporal Resolution for Studying Internal Systems at Cellular Lateral Resolution In Vivo
Translation
Microscope Body Rotation (Rotating Bodies Only)
-5° to +95°, -50° to +50°, or -45° to +45° Around Objective Focus 0.1° Encoder Resolution
Coarse Elevator Base Z (Rotating Bodies Only)
5" (127 mm) Total Travel; 1 µm Encoder Resolution
Fine Microscope Body X and Y
2" (50.8 mm) Total Travel; 0.5 µm Encoder Resolution
Fine Microscope Arm Z
1" (25.4 mm) Total Travel; 0.1 µm Encoder Resolution
gaiqing Wang
 (posted 2020-05-07 11:13:33.577)
I am looking for a cheap way to do confocal imaging in vivo. Is this Bergamo II Series Multiphoton Microscope my best option? Can you send me a quote?
YLohia
 (posted 2020-05-07 09:45:11.0)
Thank you for contacting Thorlabs. We will reach out to you directly to discuss your requirements.
jfpena
 (posted 2016-12-19 18:15:55.003)
I am looking for a cheap way to do confocal imaging in vivo. Is this Bergamo II Series Multiphoton Microscope my best option? Can you send me a quote?
tfrisch
 (posted 2016-12-22 11:44:31.0)
Hello, thank you for contacting Thorlabs. A member of our Imaging Team will reach out to you directly to discuss this system and your application.
birech
 (posted 2016-11-17 06:33:49.463)
I asked for a price quote for this product, Bergamo II Series Multiphoton Microscopes three days ago. I am working at the University of Nairobi in Kenya and would wish to order one.
Regards,
Birech
tfrisch
 (posted 2016-11-17 06:56:23.0)
Hello, thank you for contacting Thorlabs. I have forwarded this request to our Imaging Sales Team. I apologize for the delay.