Fluorescence in situ hybridization (FISH) is a modern molecular biology technique used for the detection of genetic abnormalities in terms of the number and structure of chromosomes and genes. The FISH technique is typically employed for prenatal diagnosis of congenital dementia in the Obstetrics and Genecology department. It is also routinely used to pick up qualifying breast cancer patients that are known to be highly curable by the prescription of Her2 targeted therapy. During the microscopic observation phase, the technician needs to count typically green probe dots and red probe dots contained in a single nucleus and calculate their ratio. This procedure need to be done to over hundreds of nuclei. Successful implementation of FISH tests critically depends on a suitable fluorescent microscope which is primarily imported from overseas due to the complexity of such a system beyond the maturity of the domestic optoelectrical industry. In this paper, the typical requirements of a fluorescent microscope that is suitable for FISH applications are first reviewed. The focus of this paper is on the system design and computational methods of an automatic florescent microscopy with high magnification APO objectives, a fast spinning automatic filter wheel, an automatic shutter, a cooled CCD camera used as a photo-detector, and a software platform for image acquisition, registration, pseudo-color generation, multi-channel fusing and multi-focus fusion. Preliminary results from FISH experiments indicate that this system satisfies routine FISH microscopic observation tasks.
It is widely believed that by using a digital mirror device (DMD) as the spatial light modulator (SLM) of a programmable array microcopy (PAM), it is possible to achieve a cost-effective alternative to expensive confocal imaging devices. During the past decade, the design of such a DMD based PAM instrument has been frequently reported to enhance resolution and contrast, convincing images with improved quality are rare to be seen. The concrete implementation of a DMD based PAM instrument needs to successfully resolve multiple issues such as the adverse effects caused by the tilt angle of the micro-mirrors from the base board, the registration between a micro mirror of the DMD and the image pixel of the photo-detector and so on. In this paper, we report the design of a middle body consisting of a DMD as an independent attachment to a conventional microscope to convert the latter into a confocal imaging system, in a similar way as a filter turret that is placed below the head and the objectives of a regular microscopy to convert it into a fluorescent microscopy. Images of real objects with improved contrast are provided to demonstrate the effectiveness of using a DMD as SLM to improve the contrast of a PAM instrument. Such a PAM instrument has many advantages compared to conventional laser-scanning confocal systems including lower costs and higher imaging speeds. In addition, it allows convenient dynamic adjustments between imaging quality and imaging speed.
Multispectral imaging is a powerful tool in remote sensing applications. Recently， a micro-arrayed narrow-band optical
mosaic filter was invented and successfully fabricated to reduce the size and cost of multispectral imaging devices in
order to meet the requirements for low- or mid- altitude remote sensing. Such a filter with four narrow bands is
integrated with an off-shelf CCD camera, resulting in an economic and light-weight multispectral imaging camera with
the capacity of producing multiple images at different center wavelengths with a single shot. The multispectral imaging
camera is then integrated with a wireless transmitter and battery to produce a remote sensing multispectral imaging
system. The design and some preliminary results of a prototyped multispectral imaging system with the potential for
remote sensing applications with a weight of only 200 grams are reported. The prototyped multispectral imaging system
eliminates the image registration procedure required by traditional multispectral imaging technologies. In addition, it has
other advantages such as low cost, being light weight and compact in design.
Multispectral imaging is becoming a powerful tool in a wide range of biological and clinical studies by adding spectral,
spatial and temporal dimensions to visualize tissue abnormity and the underlying biological processes. A conventional
spectral imaging system includes two physically separated major components: a band-passing selection device (such as
liquid crystal tunable filter and diffraction grating) and a scientific-grade monochromatic camera, and is expensive and
bulky. Recently micro-arrayed narrow-band optical mosaic filter was invented and successfully fabricated to reduce the
size and cost of multispectral imaging devices in order to meet the clinical requirement for medical diagnostic imaging
applications. However the challenging issue of how to integrate and place the micro filter mosaic chip to the targeting
focal plane, i.e., the imaging sensor, of an off-shelf CMOS/CCD camera is not reported anywhere. This paper presents
the methods and results of integrating such a miniaturized filter with off-shelf CMOS imaging sensors to produce
handheld real-time multispectral imaging devices for the application of early stage pressure ulcer (ESPU) detection.
Unlike conventional multispectral imaging devices which are bulky and expensive, the resulting handheld real-time
multispectral ESPU detector can produce multiple images at different center wavelengths with a single shot, therefore
eliminates the image registration procedure required by traditional multispectral imaging technologies.
Multispectral imaging is becoming a new powerful tool in a wide range of biological studies by adding spectral, spatial, and temporal dimensions to tissue abnormity and the underlying biological processes. Conventional spectral imaging systems are bulky, expensive, require multiple exposures, or extensive post-processing to align multiple images of pure spectral components. Recently, a break-through technology has emerged to instrument multispectral imaging technology into handheld real-time devices using miniaturized filter mosaic containing densely patterned micro-arrayed multiple channel bandpass optical filters. The filter mosaic can be directly placed near the focal plane, immediately in front of the imaging sensor of an off-shelf charged-coupled device/complementary metal-oxide-semiconductor camera, with the potential of one element of such a micro-arrayed filter to cover one pixel of the imaging sensor. This paper reveals the technical details of how such a micro-arrayed multichannel optical filter is fabricated using traditional multifilm vacuum deposition and the modern microlithography technologies. The selection of different coating materials, their structures, and effects to the adhesive forces between film and substrate, the spatial resolution, width of passing band, and the transmittance of the resulting miniaturized optical filter are discussed.
Multispectral imaging is becoming a new powerful tool in a wide range of biological studies by adding spectral, spatial
and temporal dimensions to tissue abnormity and the underlying biological processes. A standard spectral imaging setup
includes two major components, a band pass selection device (such as liquid crystal tunable filter and diffraction grating)
and a scientific-grade monochrome camera. Contemporary multispectral imaging technologies typically use traditional
optical filters e.g., filter wheels, a generalized Lyot filter, an electrically tunable filter, multiple-band pass filters or the
methods of dispersing light, e.g., optic-acoustic crystals. The instrumented systems are bulky, expensive, require
multiple exposures or extensive post-processing to align up multiple images of pure spectral components.
Recently a break-through technology has emerged to instrument multispectral imaging technology into handheld real-time
devices using miniaturized filter mosaic containing micro-arrayed multiple channel band-pass optical filters. The
filter mosaic can be directly placed near the focal plane immediately in front of the imaging sensor of an off-shelf
CCD/CMOS camera, with potentially one such micro-filter covers one pixel of the imaging sensor. This paper reveals
the technical details of how such a micro-arrayed multi channel optical filter is fabricated using traditional multi-film
vacuum deposition and the modern micro-lithography technologies. The selection of different coating materials, their
structures and effects to the adhesive forces between film and substrate, the spatial resolution, width of passing band, and
the transmittance of the resulting miniaturized optical filter is discussed.
Pressure ulcers have been identified as a public health concern by the US government through the Healthy People 2010 initiative and the National Quality Forum (NQF). Currently, no tools are available to assist clinicians in erythema, i.e. the early stage pressure ulcer detection. The results from our previous research (supported by NIH grant) indicate that erythema in different skin tones can be identified using a set of wavelengths 540, 577, 650 and 970nm. This paper will report our recent work which is developing a handheld, point-of-care, clinicallyviable and affordable, real time multispectral imager to detect erythema in persons with darkly pigmented skin. Instead of using traditional filters, e.g. filter wheels, generalized Lyot filter, electrical tunable filter or the methods of dispersing light, e.g. optic-acoustic crystal, a novel custom filter mosaic has been successfully designed and fabricated using lithography and vacuum multi layer film technologies. The filter has been integrated with CMOS and CCD sensors. The filter incorporates four or more different wavelengths within the visual to nearinfrared range each having a narrow bandwidth of 30nm or less. Single wavelength area is chosen as 20.8μx 20.8μ. The filter can be deposited on regular optical glass as substrate or directly on a CMOS and CCD imaging sensor. This design permits a multi-spectral image to be acquired in a single exposure, thereby providing overwhelming convenience in multi spectral imaging acquisition.
We are developing a handheld multispectral imaging device to non-invasively inspect stage I
pressure ulcers in dark pigmented skins without the need of touching the patient's skin. This
paper reports some preliminary test results of using a
proof-of-concept prototype. It also talks
about the innovation's impact to traditional multispectral imaging technologies and the fields that
will potentially benefit from it.
We report the work of developing a hand-held (or miniaturized), low-cost, stand-alone, real-time-operation, narrow bandwidth multispectral imaging device for the detection of early stage pressure ulcers.
During investigations of potential child and elder abuse, clinicians and forensic practitioners are often
asked to offer opinions about the age of a bruise. A commonality between existing methods of bruise aging
is analysis of bruise color or estimation of chromophore concentration. Relative chromophore concentration
is an underlying factor that determines bruise color. We investigate a method of chromophore concentration
estimation that can be employed in a handheld imaging spectrometer with a small number of wavelengths.
The method, based on absorbance properties defined by Beer-Lambert's law, allows estimation of
differential chromophore concentration between bruised and normal skin. Absorption coefficient data for
each chromophore are required to make the estimation. Two different sources of this data are used in the
analysis- generated using Independent Component Analysis and taken from published values. Differential
concentration values over time, generated using both sources, show correlation to published models of
bruise color change over time and total chromophore concentration over time.
Visual inspection of intact skin is commonly used when assessing persons for pressure ulcers and bruises. Melanin
masks skin discoloration hindering visual inspection in people with darkly pigmented skin. The objective of the project
is to develop a point of care technology capable of detecting erythema and bruises in persons with darkly pigmented skin.
Two significant hardware components, a color filter array and illumination system have been developed and tested. The
color filter array targets four defined wavelengths and has been designed to fit onto a CMOS sensor. The crafting process
generates a multilayer film on a glass substrate using vacuum ion beam splitter and lithographic techniques. The
illumination system is based upon LEDs and targets these same pre-defined wavelengths. Together, these components
are being used to create a small, handheld multispectral imaging device. Compared to other multi spectral technologies
(multi prisms, optical-acoustic crystal and others), the design provides simple, low cost instrumentation that has many
potential multi spectral imaging applications which require a handheld detector.
The detection and aging of bruises is important within clinical and forensic environments. Traditionally, visual and
photographic assessment of bruise color is used to determine age, but this substantially subjective technique has been
shown to be inaccurate and unreliable. The purpose of this study was to develop a technique to spectrally-age bruises
using a reflective multi-spectral imaging system that minimizes the filtering and hardware requirements while achieving
acceptable accuracy. This approach will then be incorporated into a handheld, point-of-care technology that is
clinically-viable and affordable. Sixteen bruises from elder residents of a long term care facility were imaged over time.
A multi-spectral system collected images through eleven narrow band (~10 nm FWHM) filters having center
wavelengths ranging between 370-970 nm corresponding to specific skin and blood chromophores. Normalized bruise
reflectance (NBR)- defined as the ratio of optical reflectance coefficient of bruised skin over that of normal skin- was
calculated for all bruises at all wavelengths. The smallest mean NBR, regardless of bruise age, was found at wavelength
between 555 & 577nm suggesting that contrast in bruises are from the hemoglobin, and that they linger for a long
duration. A contrast metric, based on the NBR at 460nm and 650nm, was found to be sensitive to age and requires
further investigation. Overall, the study identified four key wavelengths that have promise to characterize bruise age.
However, the high variability across the bruises imaged in this study complicates the development of a handheld
detection system until additional data is available.
This paper introduces a two-phase algorithm to extract a center-adjusted, one-voxel-thick line representation of cerebral vascular trees from volume angiograms coded in gray-scale intensity. The first stage extracts and arranges the vessel system in the form of a directed graph whose nodes correspond to the cross sections of the vessels and whose node connectivity encodes their adjacency. The manual input reduces to the selection of two thresholds and the designation of a single initial point. In a second stage, each node is replaced by a centered voxel. The locations of the extracted centerlines are insensitive to noise and to the thresholds used. The overall computational cost is linear, of the order of the size of the input image. An example is provided which demonstrates the result of the algorithm applied to actual data. While being developed to reconstruct a line representation of a vessel network, the proposed algorithm can also be used to estimate quantitative features in any 2-D and/or 3-D intensity images. This technique is sufficiently fast to process large 3-D images at interactive rates using commodity computers.