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頻譜螢光影像染色體比對系統 - SKYView EXPO

甚麼是頻譜影像技術 ( Spectral Imaging ) ?
頻譜影像技術是最先進的結合 影像 ( Imaging ) 與 頻譜 ( Spectral ) 的技術, 此技術將擷取的影像, 通過 Interferometer 以傅立葉轉換處理, 讀出影像中每一組成像素的頻譜, 經此運算處理, 因為是讀取分析頻譜, 所以, 可以用頻譜方式分辨出一般攝影機或眼睛無法分辨的影像顏色, 可以分辨出影像中任何細微差異的組成.

 

SpectraCube 安裝在顯微鏡, 

在 CDM Case Data Manager 環境下,

 使用 SKYView 分析軟體, 

包括 SKY, FISH, G-Banding 

比對分析軟體.

甚麼是 SKY ( Spectral Karyotyping ) ?

每一染色體經由染色後, 經由頻譜影像技術處理後, 皆可得到其特定的頻譜. 利用 5 組種染劑 ( FITC, Rhodamine, Texas Red, Cy5, Cy5.5 ) 依不同比率做混合雜染 ( Hrbridization ) , 我們可將人類所有染色體 ( 22 + X, Y ) 做全部的配對染色. 然而, 在一般顯微鏡下的螢光影像, 經肉眼觀察, 或用 CCD 擷取影像, 皆無法精確的分辨出每一對染色體, 因為顏色太過於接近, 容意導致誤判.

使用頻譜技術卻得到決然不同的結果, 因為是讀取每一染色體的頻譜 ( 不是螢光呈像顏色 ), 所以, 縱使肉眼觀察顏色都一樣的染色體, 我們也能精確分辨出每一染色體的詫異不同處, 連染色體異位都可精確的標定出來. 正因如此, 我們可以將頻譜的不同, 以不同的顏色指定標示, 及創造出彩色的染色體比對方法. ( SKY ).

 

 

步驟 一 步驟 二 步驟 三

上圖的染色體 ( 原始螢光影像 ) 有任何區別嗎 ? 你能精確與快速的做比對分類嗎 ?

上圖是 ' 三對 ' 不一樣的染色體.

通過頻譜影像分析, 能精確的讀出每一染色體的頻譜, 縱使原始螢光影像的顯示顏色一樣, 或者, 影像不清析, 頻譜影像皆能讀出個別的差異. 不同的頻譜曲線分佈, 可以標定出不同的顏色, 以示區別 ( 比對分析 ). 應用此技術, 可以用頻譜方式分辨出一般攝影機或眼睛無法分辨的影像顏色, 可以分辨出影像中任何細微差異的組成.
下圖為頻譜影像的 螢光雜染影像分析 ( Spectral FISH )
原始 Bright-Field 影像 一般冷卻 CCD 的攝取螢光影像. 影像的頻譜分析 通過頻譜影像分析, 能精確讀出個別的差異. 
 
     使用 SKY 發表文獻 ( 部分文獻索引 )
 
SKY 技術, 應品製倍與雜染流程, 簡易上手.

頻譜影像分析及比對, 只要經過專業的教用訓練, 都可以輕輕鬆鬆的操作此系統.

結果的精確, 值得您信賴.

SkyPaint 螢光雜染探針試劑, 目前有人類及大小白鼠所用.

必須冷藏保存.

 
 
 
 

 
SKY ( SKYView ) 內建 G-Banding 比對分析功能
Spectral FISH
Spectral FISH

SKYView 系統, 包括選配 Spectral FISH 模組

In this case, a marker was found but not identified.
Sky identified that the origin of the marker is chromosome (18).
This marker has high risk for fetal abnormalities.
Based on the SKY analysis the pregnancy was terminated.

 

Detection of a partial trisomy 21 by SKY, in a baby

with dysmorphic features and a cytogenetically normal 46,XY karyotype

 

臨床染色體比對案例 :

Human chromosome

Bone marrow

Acute Myelotic Leukemia

SKY

Translocation +(11;22)

Neuroblastoma patient

SKY 結合 Laser Micro-Dissection.
SKY

murine model system of Breast cancer

A mouse knock-out cell line for the
breast cancer susceptibility gene Brca2
was subjected to Mytomycin C treatment,
a DNA cross linking reagent. It could be demonstrated that Brca2 plays a role in the homologous recombination DNA repair mechanism, since increased non-homologous  recom-bination structures can be observed
using SKY. An important finding to understand the basic principles of neoplastic transformation in (breast) cancer

 

SKY 案例

Mouse WMP-II cell line 

 
 
SKY 比對分析圖例 :

Mouse cell line 470

Mouse chromosome SKY
Rat ( 老鼠 ) 專用 SKY-painting probes
單一染色體螢光標定 ( Human, Mouse, Rat ), Green 及 Red 兩種顏色.

ASI Dual Labeled Kit ( 標定特定染色體的 Centrometric region )

Probe : X ( Red )

Probe : Y ( Green )

SKYView EXPO 性能說明 :

Features

  • Automatic, robust color karyotyping of all sample types and kit batches.

  • Automatic accurate identification of translocations and chromosomal origins of marker chromosomes.

  • Automatic and precise classification of many FISH signals in one measurement.

  • Unsurpassed per pixel classification results.

  • G-, R-, Q- or DAPI-band images integrated with SKY images.

  • Intuitive image enhancement tools.

  • Highest quality band image enhancement.

  • Powerful spectral image analysis tools.

  • Intuitive graphic user interface.

  • Easy annotation and report tools.

  •  Images saved in common graphic formats.

 

Karyotyping Software

High resolution megapixel digital CCD camera, 1280 x 1024 spatial resolution, with 12-bit dynamic range (4096 gray scales)

Image Capture with automatic or manual exposure and contrast controls.

Image Enhancement (Contrast, Sharpening, Straightening, Rotation, etc.)

Automatic Separation of overlapping Chromosomes

Automated Counting of Chromosomes

Ability to document XY coordinates and slide label information within Acquisition Mode without having to switch back to Case Data Manager

Active, fulltime viewing of chromosomes while being repositioned within the karyotype table.

Chromosome Annotation Capabilities including text and graphics, with reference to ISCN Nomenclature of Ideograms

Annotation capabilities provided within Microsoft Paint (included), or the ability to link annotation to other programs (i.e. Photoshop, Powerpoint, etc.) of the user’s choice

Report Generation, with saving of files as *.snp filetypes for shareability.

Image Database with automated chromosome classifier G, R, Q, and DAPI Banding Patterns

Network Functionality with autonomous capability

Interface to Shire Genetics Software System: Dual direction

Compatibility to Safely Transfer Case Images between offsite computer stations with similar capability

Ability to clean and re-initialize terminals independently without powering down the network

Network capability of 25+ stations

Ability to create and print graphical statistics for department administration, including workflow, sample distribution, case distribution per technician, etc.

Description of activity history log capabilities of software:

o        Case Data Manager database maintains identification of technician or cytogeneticist working on case. 

o        Karyotype results, as well as counting results, are updated automatically in the database. Any change of the karyotyping results will open a new line entry in the results fields and documented automatically. Hence, histories of previous analysis are kept.

o        Operations done in the metaphase and karyotype are saved in the *.bnd file. Original data is always maintained! When a previous analyzed cell is loaded, previous user operations are done automatically on the original data to bring it back to its latest status.

o        History log is under consideration for development, and will be implemented if required by NYGH.

 

BandView Features

High resolution images with 12-bit digital CCD-camera (1280 x 1024 pixels)

Wide field-of-view (2/3”) for capturing widely spread metaphases in one image acquisition

Ability to join numerous frames, connecting even very sparse metaphases, including gallery of the individual frame to validate full coverage

Extremely low dark noise allowing for long exposure times

Automatically adapted to both brightfield, phase contrast and epi-fluorescence — no need to adjust the microscope illumination during the image acquisition (1 mSec to 10 Sec)

Automatically adapted to any objective magnification

Live view of image with optimal contrast is displayed throughout image acquisition

Powerful user-interface for quick, simple image acquisition

Background displayed as either white or gray values, allowing the ability to validate that no part of the image, such as  faint telomeric regions, has been missed

Inclusion of chromosome parts, such as markers, at any point in the analysis process

Availability of all tools, such as shape editing, contrast and sharpness changes, at any stage of the analysis process

Karyotyping at any stage in the analysis process, even before segmentation has been finalized

Automatic background correction, which accurately resolves highly non-uniform microscope illumination to allow for optimized segmentation

Analysis with all objective magnifications, without the need to change the C-mount or recalibrate the system parameters when the objective is changed

Superb automatic contrast to minimize effect of small debris in the metaphase

Customized settings, such as the Band Enhancement level, can be saved and applied automatically on each new image

Automatic separation of touching

Highly robust and efficient tools to correct chromosome contours and to resolve highly complex chromosome spreads

Definable “Undo” for removal of up to 10 steps in the analysis process.

10-level enhancement of fine band detail in brightfield and fluorescence

Enhance individual chromosomes, including linear and non-linear brightness and contrast controls

Restore original contrast and brightness in one click

Advanced color control for precise brightness and contrast adjustments

All original image data always maintained

Easy separation of touching and overlapping chromosomes

Automated classification of brightfield or fluorescently-labeled metaphases

Proprietary background subtraction enables automatic chromosome segmentation on sub‑optimal metaphases

Chromosome classifier libraries to improve original chromosome placement

400-, 550-, 850- band level ideograms based on ISCN‑1995 nomenclature

Robust comparison tool to simultaneously compare multiple chromosome classes

Dynamic karyotype table to fit any number of chromosomes

Multiple drag and drop for transferring and replacing chromosomes

Intuitive user interface, such as seeing the chromosomes as they are dragged in the karyotype table.

Generation of  Reports

Flexible report generation to include any combination of data, including metaphase, karyotype and case data information, physician reports and analyzed cells report

Customizable design of Report Templates to accommodate user’s requirements

Graphical Reports of Statistics: distribution of cases analyzed over time, per Technician, Physician, Specimen Type, etc.

Comparison reports of multiple cells from the same or different cases, optionally combining results of G-Band, FISH and SKY in the same report.

Flexible page layout

Easy-to-use annotation tools

Compatibility with any printer with a Windows XP driver

Customizable  workspace

Flexible use with any computer monitor —including laptops— at any screen resolution

Easy arrangement of screen windows to fit your personal work style

Practically unlimited zoom levels for any screen window

Flexible karyotype table stretches automatically to fit any arrangement, including unlimited number of chromosomes in a class

Customized karyotype view with the ability to show any or all of the following side-by-side in the karyotype: original chromosome, straightened chromosome, ideogram, banding profile, and breakpoint locations

Management of  image data

Robust image database for all ASI applications allowing viewing of all analyses of a particular case, including SKY™ and FISH.

Powerful search utility that can filter by any field, including text strings

Convenient image gallery to spot cases by metaphase or karyotype

Immediate access to all patient data in one mouse click

Direct archiving to any storage media

Extensive network capabilities for multiple systems and workstations

Upgrade to SKY™ and FISH

Seamless upgrade to SKY™ with the simple addition of an optical head — no need to purchase an additional camera

Software upgrade to FISHView™, ASI’s FISH analysis system

 

 

 

 

Applied Spectral Imaging ( ASI - 以色列 ) 產品一覽表

 

CytoLabView

微電腦控制, 數位頻譜影像技術

    請點選下列 ASI 產品資料說明 .....

BandView EXPO

G, R, Q, DAPI-Banding 染色體比對 

FishView EXPO

染色體, 細胞基因 之 FISH 影像擷取分析 

SKYView EXPO

染色體 SKY ( Spectral karyotyping ) 彩色比對, 計數及影像及資料管理中心

 

CGHView EXPO

染色體 CGH 分析

SpectralView EXPO

頻譜影像系統

Case Data Manager EXPO

影像及病例資料管理中心

 

SkyPaint probes

染色體雜染探針試劑

Single paint probes

單一特定染色體探針試劑

SpectraCube

頻譜影像擷取處理器

美嘉儀器股份有限公司 Major Instruments Co., Ltd.
台北總公司 : TEL : 02-2808-1452  FAX : 02-2808-2354  維修中心 : 02-2808-2353   高雄辦事處 - TEL : (07) 725-8177  FAX : (07) 725-8204

E-mail : major@major.com.tw    http://www.major.com.tw