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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 (New)

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

N256 Camera HeadN256 Camera Head


Features of N256 Camera

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.


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 ratios, 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 multiple ROI setting can be selected, and as a result it is possible to achieve higher frame rates. For example, ultra high frame rates such 20,000 fps @ 32x32 pixels is possible.

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




HR Camera

HR camera, which is the camera head for MiCAM02-HR, can be used with MiCAM05 processor (analog interface board is required).

HR Camera Head


Features of HR Camera

Higher spatial resolution

Spatial resolution of HR camera is varied at 92x60 - 380x244 pixels. 380x244 pixels is the highest spatial resolution among camera heads for MiCAM05.


Powerful in dark light levels

The "H-bin mode" function reduces dark noise, and it is comparable to that of a cooled-CCD camera. This function is very effective for situations of dark light or fluorescence.


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 now 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 using long term 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 results will be the same 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

Continuous output of light intensity is available as a standard function

The light intensity monitor/continuous 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, making it 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, 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 up to several hours, intermittently. Because illumination light is turned on only at the time of image acquisition, phototoxic effects to sample is minimized, and a stable baseline can be acquired.
Time LapseExample of Time-lapse imaging




High resolution camera control function (optional)

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

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 straightforward 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 from 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



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