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Neuronal Imaging MiCAM05

Overview

MiCAM05 is a high-speed imaging system that captures and visualizes small changes in fluorescence intensity from biological samples stained with fluorescent probes, such as voltage sensitive dyes and calcium dyes. MiCAM05 is designed to be the upgraded version of the previous MiCAM ULTIMA model, which has been used by neuroscience and cardiac research labs internationally, and which has been cited in many scientific research journals.

While inheriting the basic performance features of the MiCAM Ultima system, such as high-speed imaging, high S/N ratios, and intuitive operation, new features have been added in the MiCAM05 system. New features include higher spatial resolution using a newly developed CMOS camera head, long-term recording ability using high speed data transfer, synchronized imaging of up to 4 cameras, expanded functionality, and improved compatibility with external equipment.


Camera Head for MiCAM05

N256 Camera HeadN256 Camera Head

N256 Camera (New)

N256 Camera head is equipped with a new CMOS image sensor designed and developed by Brainvision.


Features

Large sensor and large pixel

N256 Image Sensor

Active Area size: 17.6mmx17.6mm
Pixel size:69μmx69μm

Applications such as membrane potential imaging require both very high speed, and the ability to detect small changes in fluorescence, such as 0.5-1%. In order to optimize S/N ratios, it is important to increase the amount of light captured by one pixel. It is easy to collect light and capture small changes using a sensor having large pixels.


Balance of high spatial resolution, high frame rates, and high S / N ratios

Number of Pixels: 256x256 pixels
Maximum Frame Rate: 1,818 fps (at 256x256 pixels)
Dynamic range: 71dB

The overall performance has been improved in a balanced manner. This includes maximum frame rate, spatial resolution, and S/N ratio, all of which are important factors for high-speed membrane potential imaging.


Wide dynamic range mode and high sensitivity mode by adjustable well depth

Wide dynamic range mode (D-mode): 3,000,000 electrons
High sensitivity mode (S-mode):    600,000 electrons

It is now possible to easily switch the full well capacity of the camera in the acquisition software. Two modes are available: wide dynamic range mode (D-mode), and high sensitivity mode (S-mode). D-mode is used for tissue samples with high fluorescence light intensity, and S-mode is used for cultured cell monolayers or preps with low fluorescence light intensity. This adjustable well depth makes it possible to image a variety of biological samples using a single camera.


ROI readout for higher frame rates

Preset partial readout setting and multi ROI setting can be selected, and as a result it becomes possible to achieve higher frame rates. For example, extremely high frame rates such 20,000 fps @ 32x32 pixels is possible.

      All pixels readout (256x256 pixels)       multiple ROI readout (32x32 pixels x 9 area)







New Features in MiCAM05 Processor

Direct data saving and long-term data acquisition with USB3.0 high-speed data transfer

New USB3.0 interface allows for significantly faster data transfer from processor to PC. It is possible to save data directly to HDD or SSD, and long-term recording of several minutes up to a few hours can be achieved, regardless of the RAM capacity (Note that sampling rate, number of pixels, number of cameras used and PC specifications will affect the total recording time).


Approximate number of recordable frames and recordable time
(Acquisition with single camera and long time recording mode)
Disk Capacity Pixels Frame Rate
(fps)
Number of
recordable frames
Recordable time
(min)
SSD 1TB 256x256 500 6,840,000 228.0
1,000 6,840,000 114.0
1,818 6,840,000 62.7
128x128 500 27,360,000 912.0
1,000 27,360,000 456.0
5,263 27,360,000 86.6

(*Approximate number of recordable frames and recordable time shown above are averaged values that were measured using the PC configuration which is included with MiCAM05. There is no guarantee that the same results can be in all circumstances.)


Up to four cameras can easily be connected and used in a completely synchronized multi-camera system

Up to two camera interface boards can be connected to the MiCAM05 processor. Each interface board is equipped with two camera ports, therefore up to four of the same type of camera head can be connected at any time. This allows for simultaneous imaging of multiple fluorescent wavelengths as well as three-dimensional imaging from different angles.

Camera Port

Real time light intensity monitor/output is available as a standard function

The real time light intensity monitor/output function, available as an optional component of MiCAM Ultima (DAC/ADC expanding unit), is available as standard equipment with MiCAM05. This function is designed to output the light intensity changes of up to eight user selected pixels via eight BNC connections. Therefore, waveforms of fluorescence change can be monitored and output before, during, and after data acquisition. It is also possible to monitor the change in light intensity in real time, using optional software. Real time light intensity monitorLight intensity outputs from eight BNC connections      Monitor with an oscilloscope
Real time light intensity monitorWave monitor on software



Various I/O connectors, and improved compatibility with external devices

MiCAM05 has multiple input and output connections available, in comparison to previous MiCAM systems. These universally standard I/O connections can be synchronized with image acquisition and external devices. An additional slot has been added to MiCAM05 so that customization is also available, depending on the research application.

Panel

Automatic light control and adjustment of light intensity

In addition to the “Shutter” output terminal for controlling the timing of illumination in synchronization with image acquisition, there is a “LightCtl” signal output terminal for automatically adjusting the intensity of the excitation light. In this way, light intensity or saturation percentage of the image captured by the camera, can be adjusted using the software. As the background light intensity becomes similar throughout an experiment and even for different biological preps, the experimental repeatability can be improved.

light control


Other Features

Turnkey system ready for immediate use

The system consists of hardware/software specifically designed for high-speed fluorescence imaging, and the PC and monitor are included as standard equipment. Before use, calibration/setting/tuning is not necessary. The hardware connection is straightforward between components using cables. It is possible to acquire data immediately after setup.

Diagram

Pre/Post trigger mode

MiCAM05 can record activity which occurs before and after a trigger signal is generated. When an external trigger occurs such as a spontaneous biological signal, or a trigger pulse from an external stimulator, etc..., a specified amount of "pre-trigger" data can be saved. The "post-trigger" data is also collected in the same recording. As a result, both pre-trigger data and post-trigger data can be saved within one acquisition period.


Time lapse imaging

MiCAM05 can capture phenomenon that changes in a few minutes to several hours, intermittently. Because illumination light is turned on only at the time of image acquisition, it is expected that photodamage to sample is minimized, and a stable baseline of acquired data can be obtained.Time LapseExample of Time-lapse imaging




High resolution camera control function (optional)

It is possible to control a high resolution camera with 1920x1200 pixels (BV-XB1 camera). High resolution imaging can be applied to various applications including detection of intracellular Ca 2+ dynamics, flavoprotein fluorescence, various FRET imaging, and also imaging of animal behaviors. High resolution imaging with BV-XB1 can also be synchronized with in-vivo high-speed imaging using MiCAM05.

High resolution camera    Example of flavoprotein imaging

   BV-XB1                      Example of flavoprotein imaging


Main Features of Software

BV Workbench

BV Workbench is the image acquisition software that controls MiCAM05 imaging system. In addition to image acquisition and control of inputs and outputs of the imaging system, analysis functions including movie playback, wave display, filter functions, etc…are also included. BV Workbench has an intuitive user interface that is easily operated with the use of a mouse.

BV WorkbenchBV WorkbenchAcquisition Setting                                         Monitor windows and wave display

BV Workbench    BV WorkbenchData Analysis                                                    2 image overlay

Features
  • Synchronized control of multiple imaging systems and electrical stimulator
  • Image acquisition mode: Single acquisition (Save to PC memory or direct recording to SSD/HDD), Averaging, Time Lapse Imaging
  • Image overlay, Image Flip/Rotation
  • Wave display of light intensity change of pixel (real-time or after image acquisition)
  • Light intensity histogram
  • Filtering (Gaussian, Mean, Median, Inverse polarity)
  • Tools (alignment correction、illumination correction, binning, crop, time range)
  • Graphic Diagram of simulation pulse pattern


BV_Ana

BV_Ana

BV_Ana is the software used to analyze optical imaging data. It was developed mainly for analysis of neuronal activity and cardiac activity using voltage sensitive dyes.

By superimposing the change in light intensity of each pixel onto the background image, the location of light intensity changes as well as the percentage of light intensity change can be easily analyzed. It is also very easy to create presentation materials in the form of images and movies.

 Tile Image DisplayTime Differential Processing4 image simultaneous display Tile Image Display                 Time Differential Processing                       4 image simultaneous display Isochronous maps    Isochronous maps    Multi Wave Display Isochronous maps          Isochronous maps             Multi Wave Display

Features
  • Innovative software capable of intuitive operations
  • Bleaching/Drift curve correction functions
  • Enhanced processing functions such as high-pass/low-pass filters, fast fourier transform function
  • Ability to import 16bit gray-scale multi-TIFF format data other imaging systems
  • Export data such as images and movies, which can be used as presentation materials



Applications

  • High speed membrane potential imaging using voltage-sensitive dyes
  • High speed calcium imaging
  • Intrinsic fluorescence imaging based on flavoproteins and hemoglobin
  • Imaging FRET, GCaMP, and various fluorescent proteins such as VSFP
  • Ratiometric fluorescence imaging with multiple camera heads
  • 3D or panoramic imaging with multiple camera heads
  • Detection of small changes in light intensity at high speeds

Sample Data



Specifications

System
Name Multifunctional High Speed Imaging System
MiCAM05-N256
Model MC05-N256
Standard System
Configuration
Camera Head (1-4), Processor, PC, Monitor,
Acquisition software, Analysis Software
Supported OS Windows 7/10 64bit
Default/Optional Function
Single Trial Acquisition Default
Long Time Acquisition
(direct saving to disk)
Default
Averaging Acquisition Default
Multiple ROI Readout for
Higher Frame Rate
Default
Time-Lapse Acquisition Default
High Resolution Camera
Control
Option (sold separately)
Acquisition software Default
Analysis software Default
Camera Head
Name N256 camera
Model N256CM
Image Sensor Original CMOS
Active Pixels (H x V) 256 x 256
Active Image Area (H x V) 17.6mm x 17.6mm
Pixels Size (H x V) 69μm x 69μm
Shutter Mode Global Shutter
Maximum Frame Rate and
Pixels (for 1 camera)
1,923fps@256x256 pixels
3,125fps@192x192 pixels
5,556fps@128x128 pixels
20,000fps@32x32 pixels
Quantum Efficiency 55%@550nm / 50%@600nm
Well-Depth (switching) 3,000,000 e- / 600,000 e-
Dark noise 500 e- / 130 e-
Dynamic range 71dB
Lens mount M42 mount (Option: C or F mount)
Interface Special Cable
Dimension (WxHxD) 80mm x 80mm x 50mm
Weight 474g
Processor
Name MiCAM05 processor
Model MC05PRC
Camera Interface 1 (up to 2)
Camera Port 2 (up to 4)
Slot for Optional Unit 1
Buffer Memory
Inside Processor
4GB (up to 8GB)
Analog Inputs for
synchronous recording
4ch
Output for Stimulation Pulse 2ch
Input for external trigger signal 1ch
Output for Lighting Control 1ch
Input/Output for Frame Timing 1ch
Output for Real Time
Light Intensity Monitor
8ch
Output for Light Intensity Control 2ch
Current Output 2ch
Output for Acquisition Status 1ch
Output for Trigger Wait Status 1ch
Digital I/O port 1 (25 pins)
Trigger Mode Software, TTL External Input,
Analog External Input, Usage of Optional Unit
Data Transfer Mode USB3.0 (for basic functions)
USB2.0 (for Real Time Light Intensity Monitor)
Recording Mode Temporary storage to PC RAM /
Direct data saving to PC drive (SSD/HDD)
Maximum Recordable
Frame Number
Dependent on camera used and image acquisition settings
Power Supply AC100V-230V、50Hz/60Hz 150W
Fuse T2A L 250V
Dimension 430mm (W) x 350mm(D) x 75mm(H)
Weight 6.2kg

* Specifications and appearance are subject to change without prior notice due to continuous developments and improvements.






Selectable Number of Pixels and Maximum Frame Rate (1 camera)
Selectable Number of Pixels (HxV) Maximum Frame Rate (fps)
NR Mode OFF NR Mode ON
2 Times Averaging
NR Mode ON
4 Times Averaging
256x256 Full Resolution 1,818 1,923 1,031
192x192 Square ROI 2,941 3,125 1,695
128x128 5,263 5,556 3,333
64x64 12,500 12,500 8,333
256x128 Horizontally Long
Rectangle ROI
3,333 3,448 1,887
256x64 5,882 5,555 3,226
256x32 9,091 8,333 5,000
128x256 Vertically Long
Rectangle ROI
3,030 3,333 1,852
64x256 4,348 5,000 3,125
32x256 5,556 7,143 4,762
32x32 16 blocks 2,381 2,632 1,724
15 blocks 2,500 2,778 1,818
14 blocks 2,703 2,941 1,961
13 blocks 2,857 3,125 2,083
12 blocks 3,125 3,448 2,273
11 blocks 3,333 3,704 2,439
10 blocks 3,704 4,000 2,703
 9 blocks 4,000 4,348 2,941
 8 blocks 4,545 5,000 3,226
 7 blocks 5,000 5,556 3,704
 6 blocks 5,556 6,250 4,167
 5 blocks 6,667 7,143 5,000
 4 blocks 7,692 8,333 5,882
 3 blocks 10,000 11,111 7,692
 2 blocks 12,500 14,286 10,000
 1 blocks 20,000 20,000 16,667


Selectable Number of Pixels and Maximum Frame Rate (2 camera)
Selectable Number of Pixels (HxV) Maximum Frame Rate (fps)
NR Mode OFF NR Mode ON
2 Times Averaging
NR Mode ON
4 Times Averaging
256x256 Full Resolution 1,087 1,064 1,031
192x192 Square ROI 1,818 1,754 1,695
128x128 3,704 3,571 3,333
64x64 9,091 8,333 8,333
256x128 Horizontally Long
Rectangle ROI
2,041 1,961 1,887
256x64 3,571 3,226 3,125
256x32 5,882 5,000 5,000
128x256 Vertically Long
Rectangle ROI
2,000 1,961 1,852
64x256 3,571 3,448 3,125
32x256 5,556 5,556 4,762
32x32 16 blocks 1,887 1,613 1,613
15 blocks 2,041 1,724 1,724
14 blocks 2,174 1,852 1,852
13 blocks 2,326 1,961 1,961
12 blocks 2,500 2,128 2,128
11 blocks 2,703 2,326 2,326
10 blocks 2,941 2,500 2,500
 9 blocks 3,226 2,778 2,778
 8 blocks 3,571 3,125 3,125
 7 blocks 4,000 3,448 3,448
 6 blocks 4,762 4,000 4,000
 5 blocks 5,556 4,762 4,762
 4 blocks 6,667 5,556 5,556
 3 blocks 8,333 7,143 7,143
 2 blocks 11,111 10,000 10,000
 1 blocks 16,667 16,667 16,667


Discontinued Models
System
Name MiCAM05-Ultima MiCAM05-D225
Model MC05-UL MC05-D225
Camera Head
Name Ultima camera D225 camera
Model CAM-ULT D225
Image Sensor Original CMOS Original CMOS
Active Pixels (HxV) 100x100 (fixed) 240x256 (fixed)
Active Image Area (HxV) 10.0mmx10.0mm 7.2mmx7.7mm
Pixel Size (HxV) 100μmx100μm 30μmx30μm
Shutter Mode Rolling Shutter Global Shutter
Maximum Frame Rate
(for 1 camera)
10,000fps 1,000fps
Well-Depth 1,500,000 e- 500,000 e-
Lens Mount C mount C mount
Dimension (WxHxD) 80mmx65mmx50mm 75mmx65mmx50mm
Weight 270g 246g
Processor
(Same as MiCAM05-N256)

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