DLM Confocal Microscope™


Innovate...

Aeon Imaging’s Digital Light Microscope (DLM) is a cost-effective and fully customizable confocal microscopy
platform built using Edmund Optics® or Thorlabs® components.

Innovate with the tools you need:

  • Control the microscope and stream image data using Firefox, or Chrome browsers over your local area network
  • Explore new imaging frontiers by learning the ins and outs of the DLM hardware through our white pages and datasheets
  • Push the limits by purchasing Aeon’s software development kit, which provides complete access to the source code and software documentation

Features

Using Aeon’s DLM, customized and modular imaging systems can be easily built for bright-field, dark-field, differential contrast, and fluorescence imaging. Get the DLM essentials at an outstanding price of $4,295. Then pick from thousands of compatible building-blocks offered by Edmund Optics® and Thorlabs® and build the latest imaging technology into your microscope.

Not ready to go it alone? Aeon Imaging has developed starter microscope kits to get you up and imaging. Customize your microscope, receive the parts, and assemble using our complete step-by-step instructions.

How Does it Work?

Our patented DLM technology uses a digital light projector for target illumination instead of the more traditional laser source and scanning element(s). The projector rapidly illuminates the target with a sequence of lines that is directly imaged (i.e. without descanning) onto a CMOS sensor with rolling shutter detection. By spatially and temporally synchronizing the illumination lines to the CMOS rolling shutter, the rolling shutter acts as a flexible electronic confocal aperture that can be adjusted in width and position with pixel-level accuracy from frame-to-frame.

Check out all the details in our Technology section on this page.

Explore…

Popular applications of the DLM microscope:

Aeon Confocal Microscope from the topFluorescence Microscopy
Aeon’s DLM seamlessly switches between red, green, or blue illumination
LEDs. With the addition of excitation and emission filters, sample fluorescence
can be readily measured.
 
 
 
 
Images taken by the Aeon Confocal MicroscopeImages taken with the Aeon Confocal MicroscopeDifferential Contrast Microscopy
Achieved by alternating the confocal aperture width every other frame through software. By subtracting a narrow aperture image from a wide aperture one, image edges are emphasized, creating an effect similar to differential interference contrast microscopy.
 
 
 
 
Images taken by the Aeon Confocal MicroscopeImages taken with the Aeon Confocal MicroscopeMagnification with 3rd party objectives
Users may insert their own microscope objectives when imaging in epi-illumination mode to achieve high magnification.
 
 
 
 
 
Fibroadenoma of the BreastFibroadenoma of the BreastFibroadenoma of the BreastFibroadenoma of the BreastBiomedical Applications
Images of Fibroadenoma of the breast taken using a 1X objective.

 
 

More images taken with the DLM

Discover… Novel DLM Technology:

Aeon Imaging has developed a unique approach to imaging that uses a digital light projector as an illumination source and CMOS rolling shutter detection. The projector rapidly illuminates the target with a sequence of lines that is directly imaged (i.e. without descanning) onto a CMOS rolling shutter sensor. By spatially and temporally synchronizing the projected lines to the position of the rolling shutter, spatial filtering is obtained.

Aeon’s DLM technology combines some of the best features present in Confocal Scanning Laser and Structured Illumination Microscopy:

  • Cost-Effective:
  • A digital light projector is used instead of a laser source and scanning mirrors.
  • Flexible:
  • Precise confocal aperture adjustments can be made electronically on a frame-to-frame basis. Custom illumination patterns can be easily uploaded to the projector.
  • Fast:
  • Real-time frame rates without the need to acquire and process 3 or more structured illumination frames to achieve optical depth sectioning
  • Easy to Align:
  • The DLM performs fine optical alignment automatically by adjusting the sensor’s region of interest
  • Modular:
  • Add and remove standard Edmund Optics or Thorlabs optical components in minutes.
  • Versatile:
  • Adjust the rolling shutter in real-time to enable bright field, dark field, differential contrast and enhanced axial resolution imaging. Image with red, green, or blue illumination.

How does it work? First, consider CMOS Rolling Shutter Detection

sensor

Unlike global shutter CCD sensors, which integrate charge evenly across the entire active sensor area during the exposure time, the rolling shutter method of detection, commonly found in CMOS sensor chips, progressively integrates light over only one or more pixel rows at a time, spanning a “shutter width.” Light incident on the pixel array outside the active rolling shutter read area is not captured, and therefore cannot reduce contrast. Unlike a mechanical pinhole, the width and position of the rolling shutter can be electronically adjusted in real-time at pixel-level accuracy. Electronic aperture control is robust, fast and accurate, which greatly enhances the ability to switch between bright and dark field imaging modes, and to optimize the trade-off between the amount of spatial filtering and light detected.

Next, consider state-of-the-art DLP® technology

Near Infrared digital light projectorDigital micromirror devices (DMD) comprise an array of individually addressable MEMs mirrors that can each be set to one of two angular positions. When interfaced with high power light sources and projection optics, and driven using a standard video input, these devices are commonly known as digital light projectors (or digital light processors). The DLP LightCrafter®, provided by Texas Instruments®, is a handheld and versatile example of modern integrated DLP® technology. The LightCrafter can operate as a standard video projector, with 8-bit depth RGB color output at 60Hz. Each frame is displayed by rapidly projecting its composite "bit-planes" in sequential time slots. A bit-plane exists for each digital bit of resolution for each of the red, green, and blue illumination channels (i.e. there are 24 bit-planes displayed per 60Hz frame). Unique to Texas Instruments' DLP technology, the projector can be operated in a "structured light mode", whereby the number of colors and bit depth can be reduced to achieve a higher frame rate. For example, the LightCrafter can accept 3-color, 8-bit 60Hz video input and display monochrome, single bit-plane output patterns at 1,440Hz. To achieve up to 4kHz pattern display rates, up to 96 bit-plane patterns can be pre-loaded into the LightCrafter.

A complete description of Texas Instruments’ DLP LightCrafter and its capabilities can be found here.
 

Aeon Imaging’s DLM technology

Aeon Essentials Aeon Imaging’s DLM Essentials Kit includes:

  • DLP LightCrafter pre-configured for structured light mode using stored line patterns
  • The necessary optics to collimate and focus the projected light patterns onto a target
  • Single-board computer with a 1GHz ARM processor
  • CMOS rolling shutter sensor
  • The line patterns and projector collimation optics match the illumination and detection fields of view, while the electronics provide temporal synchronization. Confocal imaging has never been so easy! Customize your microscope by relaying the imaging plane to the target and sensor, add microscope objective, polarizers, or wavelength filters as-needed from Edmund Optics’ or Thorlabs’ extensive catalogues, and align your system through a standard internet browser in minutes.

    Envision...

    The many possible configurations and designs.

    10x light Reflectance DLM with Thorlabs® Component
    Aeon Confocal Microscope from the top Side view of the DLM-1x Confocal Microscope Aeon Confocal Microscope from the top Aeon Confocal Microscope from the top

     

    10x light transmission DLM with Thorlabs® Component
    Aeon Confocal Microscope from the top Side view of the DLM-1x Confocal Microscope Aeon Confocal Microscope from the top

     

    1x light transmission DLM with Edmund Optics® components
    Aeon Confocal Microscope Side Photo of the DLM-1x Confocal Microscope Aeon Confocal Microscope from the top

     

    1x light transmission DLM with Thorlabs® Components
    Aeon Confocal Microscope from the top Side Photo of the DLM-1x Confocal Microscope

     

    Aeon Essentials Kit
    Aeon Essentials

    Perform ... Axial Depth Sectioning

    Aeon Imaging’s DLM Essentials Kit is pre-configured for confocal imaging using a sequence of single line patterns that collectively illuminate the field of view. By narrowing the shutter width, defocused light from outside the focal volume is removed from the image.

    Brightfield View

    Brightfield Image of Plant Leaf

    Minus sign

    Darkfield Image

    Darkfield Image of Plant Leaf

    Equals sign

    Subtracted Image

    Subtracted Image of Plant Leaf

    Rather than image with very narrow aperture widths (e.g. a shutter width of less than 10 pixels), which can cause ‘light-starved’ conditions, similar axial depth sectioning can be achieved using dual line illumination patterns. This configuration, which can be easily set in the Advanced Settings of the DLM user interface, loads a sequence of line-pairs into the DLP LightCrafter. Once the dual-line patterns are loaded, the camera is set up to acquire a pair of images and compute their difference.

    The primary image has the rolling shutter synchronized with one of the projected lines, as in standard imaging. The secondary image positions the rolling shutter between the projected line pairs, with a shutter width that minimizes the light return from an in-focus target. When the difference between primary and secondary images is computed, the light returning from in-focus regions of the target will be minimally affected, while undesired light from outside the focal volume is rejected.

    To illustrate the effectiveness of this approach, the point-spread function was measured using Aeon Imaging’s DLM kit, set up in reflectance mode with a 10X 0.25NA objective. The blue bright-field point-spread function shows the depth sectioning achieved with a 20 pixel shutter width. The green dark-field point spread function shows the intensity profile obtained when the shutter is placed between the projected line patterns. The red subtracted image substantially narrows the point-spread function, and enhances the optical slicing capability of the microscope.
    A “z-stack” series of bright-field and subtracted images of a leaf target is shown below. At a single depth position, the image of the leaf in bright-field (A), dark-field (B), and subtracted (C = A-B) is shown. As expected, the dark-field image is comprised of unwanted out-of-focus light that is removed during image subtraction.

    Focus...

    On the capability and versatility of the DLM.

    Aeon Imaging’s DLM Essentials Kit (photo) includes a Texas Instruments® DLP LightCrafter as an illumination source. Details about how we use this component can be found in our technology section. Full details about its capabilities and functionality can be found here.

    Illumination Source (DLP® LightCrafter™) Specifications
    Illumination Channels Red Green Blue
    Center Wavelength 630nm 540nm 468nm
    Approximate Bandwidth 20nm 80nm 25nm
    Native Resolution
    608x684 pixels
    Maximum Number of Pre-Loaded Monochrome 1-bit Patterns
    96

    IR Spectrum for a DLP® LightCrafter™ A representative spectrum of the DLP® LightCrafter™ output is shown (left). The spectrum was captured using an Ocean Optics® USB4000-VIS-NIR spectrometer.

     

     
    Aeon Imaging’s DLM Essentials Kit (photo) also includes a monochrome Aptina™ 5MP CMOS sensor with an electronic rolling shutter. Details about how we use this component can be found in our technology section.

    Sensor Specifications (Aptina™)
    Active Pixels 2592H x 1944V
    Pixel Size 2.2 x 2.2μm
    Maximum data rate / master clock 96 Mp/s at 96 MHz
    Responsivity 1.4 V/lux-sec (550nm)
    ADC resolution 12-bit, on-chip *
    Pixel dynamic range 70.1dB *
    SNRMAX 38.1dB *
    *Note: In order to maximize our streaming video rates, we use only the top 8 bits of digital resolution. Please contact us if you would like you would like a DLM kit configured for 12-bit digital resolution.

    Aeon Imaging’s DLM Essentials Kit (photo) provides both spatial and temporal synchronization between the illumination and detection fields of view. With the addition of magnification using commercial microscope objectives, the field of view can be reduced and higher optical resolution obtained.

    Native DLM Specifications (as shipped, without additional optics):
    Projector Pixels per Line Pattern 2
    Projector Monochrome 1-bit Pattern Rate 2.62kHz
    Projector Maximum Average Power at the Target* 95μW (red); 160μW (green); 130μW (blue)
    Field of View 4.51mm x 3.38mm (H x V)
    Detector Active Pixels 2048H x 1536V
    Binning 2X (streaming images have 1024 x 768 pixels)
    Sensor Frame Rate 27.3Hz
    Monochrome Streaming Video Rate (on a desktop connected using a cross-over Ethernet cable) 10Hz
    * Higher output power is possible with active cooling. Please contact us if your require specifications outside these ranges. We’ll be happy to work with you to try to find a solution.

    Optical components can be added to relay or magnify the field of view. The achievable resolution, and any aberrations or distortion, is highly dependent on the quality of the optics used and their alignment. To help new users explore the capabilities of the DLM, we provide the parts list and assembly instructions for a confocal microscope kit.

    DLM 1X Relay Lens Confocal Microscope Kit Specifications:
    1x Imaging Resolution on 1951 USAF Test Target 8.8μm
    Illumination Line Width at Target 35.2μm
    Minimum Shutter Width at Target 2.2μm (1 CMOS pixel)*
    Field of View 4.51mm x 3.38mm (H x V)
    * Imaging performance is best when the shutter width is at least 20 pixels wide.

    Representative 1951 USAF Resolution Test Chart Images:

    Figure 1.  
    Figure 1 1X Relay Lens Transmission DLM. Group 5, Element 6 bars are resolvable, which have a 8.77μm line spacing.
    Figure 2.  
    Figure 2 Reflectance-based DLM with a 10X 0.25NA Olympus® Plan N Achromat Objective. Group 9, Element 1 bars are resolvable, which have a 0.98μm line spacing.