ABSOLUTE COUNTING OF PROTEINS SUSPENDED IN A THIN CELL USING HIGH RESOLUTION IMAGING TECHNOLOGY
The Occhio FC200SHR is part of a family of highly sophisticated imaging systems, that are byproducts of a series of collaborative studies in light, imaging, and sample transport, in connection with the University De Liege in Belgium.
The FC200SHR IPAC, in particular, was developed to solve some of the current technological hurdles with protein aggregate counting.
LIMITATIONS OF EXISTING TECHNOLOGY
SUB MICRON DETECTION OF AGGREGATES
LIQUID SUB SAMPLING
AGGREGATE DIFFERENTIAL SPEEDS
HIGH RESOLUTION IMAGING TECHNOLOGY
This high resolution imaging technology, is accomplished in part, by using special backlighting techniques focused from behind the sample. An adjustable range, monochromatic, blue light source has been shown to provide optimal contrast conditions for the particles, and virtually eliminate diffraction. The light is collimated, and acts as a high definition caliper. (Fig. 1.2) In the case of liquids and suspensions, the light flashes simultaneously with a high speed, high definition camera, for extremely robust particle definitions.
COMPARISON SLIDES OF OPTICAL MICROSCOPY AND NEWLY DEVELOPED DIGITAL IMAGING TECHNIQUES
SUB MICRON DETECTION OF AGGREGATES-In order to improve the quality of the images, the Occhio FC200S HR uses a higher resolution camera, with an optical calibration of .38 microns per pixel, compared with an estimated 1.4 microns per pixel, on a Brightwell DPA 4200. The flow cell on an Occhio FC200S HR, is also half the width at 50 microns.
COMPARISON USING LATEX SPHERES
SPECIAL SYRINGE PUMP FOR PRECISE SAMPLING.
SOLUTIONS TO TECHNOLOGY LIMITATIONS
SUB-MICRONIC DETECTION OF AGGREGATES
DECREASE THE FLOWCELL THICKNESS TO 50 MICRONS (HALF OF STANDARD FLOW CELL THICKNESS)
INCREASE OPTICAL CALIBRATION TO 38 MICRONS PER PIXEL
ENABLE GHOST (TRANSPARENT) PARTICLE DETECTION
LIQUID SUB SAMPLING
USE AN ABSOLUTE SPACER
HIGH SPEED CAMERA
PRECISE PUMPING WITH NEW SYRINGE PUIMP
AGGREGATE DIFFERENTIAL SPEED
AGGREGATE TRACKING ALGORITHM
OCCHIO FC200SHR PARTICLE CHARACTERISTIC ANALYZER FOR PROTEIN AGGREGATES
SUB VISIBLE PARTICLE AND AGGREGATE DETECTION TO 200 nm
OCCHIO FLOWCELL TYPE PARTICLE ANALYZER FOR LIQUIDS AND SUSPENSIONS WITH SYRINGE PUMP AND HIGH RESOLUTION CAMERA OPTION FC200S HR
COLLIMATED LIGHT SOURCE ACTS LIKE A CALIPER TO PROVIDE SUPERIOR PARTICLE DEFINITION IN REAL TIME
IMAGE MAGNIFICATION IS INDEPENDENT OF THE OBJECTS DISTANCE OR POSITION IN THE FIELD OF VIEW
OCCHIO DIGITAL IMAGE ANALYZERS HAVE THE ABILITY TO DETECT AND QUANTIFY BOTH PARTICLE SIZE AND PARTICLE SHAPE, PAVING THE WAY FOR THE ADVANCED ANALYSIS OF PARTICLES IN BOTH LIQUID AND SOLID FORMS.
CORRELATIONS TO OTHER PARTICLE SIZE ANALYSIS METHODS, LIKE SIEVE ANALYSIS AND LASER DIFFRACTION, ARE SEAMLESSLY EMPLOYED. EVEN SHAPE PARAMETERS LIKE LENGTH, FLATNESS, AND ANGULARITY IN AGGREGATES, OR THE LENGTH OF STRINGY PARTICLES LIKE SHREDDED CHEESE, OR TOBACCO, ARE POSSIBLE.
ONCE A BASIC CORRELATION HAS BEEN ESTABLISHED, THE RESULTS ARE FINE TUNED WITH THE OCCHIO CORRELATION PROGRAM, ALLOWING FOR TURNKEY SOLUTIONS TO PARTICLE CHARACTERIZATION.