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Fluorescence In Vivo Endomicroscope 2nd generation

The flflexible hand-held probe of the FIVE2

images can be captured at any angle with sub-micron resolution

Image resolution is 0.55?m lateral and 5.1?m Axial

vary image depth in the Z-Axis:up to 400 microns(tissue dependent)

Thin optical sections enable reconstruction of 3D tissue structure in remarkable detail



Detail

Overall Introduction

The hand-held OptiScan FIVE2 system is the most advanced miniaturised confocal endomicroscope in the world.The FIVE2 enables you to push the boundaries of your research with maximum ?exibility, viewing tissue at any angle with sub-micron resolution.


FIVE 2-3.png



Product Principle

Optiscan technology is easy to use. Following application of a fluorescent dye, simply touch the probe against the tissue of interest and an image will be displayed on the screen in front of you.


A laser light passes through an optical fibre to a hand-held probe. Optics in the probe focus the laser to a diffraction limited spot in the tissue. Fluorescence is excited at this spot and the fluorescent light is captured into the optical fibre and passed to a detector. An image is generated by scanning the focussed spot through an image plane, then compiling the point intensity measurements. The image plane can be translated in depth, allowing generation of 3D image stacks.


Optiscan 原理.png


Product Features

1.Slide-free and biopsy free:users can generate images at the sub-cellular level,in real-time.

2.Viewing live biological systems in-vivo at the cellular level in their entirety:not achievable through other imaging techniques.

3.Non-destructive:image creation is non-destructive to the tissue of interest.

4.Portable:easy to work on live tissues.

5.Image depth variation:unique ability to vary image depth in the Z-Axis:up to 400 microns(tissue dependent).

6.Zero cost image acquisition:just staining cost required.

7.Post-processing versatility:images can be easily post-processed to produce a range of presentations(incl 3D)


Why Choose ViewnVivo??

  1. Superb resolution:Using a distal end single scanning fibre rather than a proximally placed scanning fibre bundle provides unmatched lateral and axial resolution. The single fibre eliminates dead spaces and broken fibres, delivering high sensitivity and long lifespan.

  2. Never miss an image:The ViewnVivo? imaging software includes an Image Rollback function. Users can select and save images from up to 60 memory held frames. Individual images or image selections can be saved or made into movie files.

  3. Longitudinal studies:High level disinfection and sterilisation options provide a cost-effective, practical way to enable longitudinal studies on the same subject, as well as the ability to image cell cultures and other samples where sterility is required.

  4. Long-life probes:ViewnVivo? probes are not a consumable item and will retain their imaging quality through years of operation and hundreds of procedures

  5. Precise control in the Z-dimension:ViewnVivo? reports the position of the focal plane with micron accuracy. The imaging software provides an intuitive interface to set and collect Z-Stacks. This data can be turned into movies or ported into Fiji for processing

  6. Image from any angle:The handheld flexible probe allows the user access to previously inaccessible samples.




Wide Range of Applications

Cancer research

?Tumour development/regression 

?Metastasis 

?Tumour detection/diagnosis 

?Border demarcation


Tissue regeneration

?Stem cell research 

?Orthopaedic tissue engineering 

?Scaffold  imaging 

?Tissue rejection


Cell tracking

?Stem cell research 

?Inflammatory responses 

?Cell-cell  & cell-tissue interactions


Photodynamic Therapy (PDT)

?Photosensitiser distribution 

?PDT response 

?Lesion detection & identification (PDD)


Infection & treatment

?Tissue microflora &microbicide response


Pharmacology

?Drug delivery & pharmacokinetics 

?Tissue response


Pathology

?Acute lung injury & animal models 

?Liver disease models 

?Morphological changes in disease


Microvascular research

?Dynamics and structure 

?Angiogenesis/anti-angiogenesis 

?Thrombosis


Animal models

?Tissue injury & disease


Molecular Imaging

?Receptor labelling 

?Protein expression profiling 

?Transgenic animals (eg GFP/YFP)



Application

1、Oral Cancer

Left: Normal

Middle: Pre-cancer

Right: Cancer

Optiscan technology captures sub cellular detail of oral mucosa in real time without having to do a biopsy. Scale bar =100um


1. 口腔癌.jpg

2、Cervical Cancer

Left: Healthy squamous epithelium

Right: Cervical intraepithelial neoplasia

0.05%% acriflavine solution applied topically. Scale bar = 100um

Left: healthy colon

Right: Adenocarcinoma

Contrast agent: IV fluorescein sodium


3. 宫颈癌.jpg


3、Tumour angiogenesis

Left: Normal dermal microvasculature

Right: Melanoma-affected dermal microvasculature

In vivo CLE imaging shows changes in melanoma-associated vasculature in and around an implanted melanoma tumourof an athymic mouse. lntravenous administration of (0.3 ml, 10 mg per ml) FlTC-dextran was used as contrast agentscale bar = 100um


6. 肿瘤血管生成.jpg


4、Autofluorescence

Left: orderly collagen fibres in ACL

Right: disordered collagen fibres after repeat loading of ACLAutofluorescence imaging of the anterior cruciate ligament (ACL). This study looks at ACL Failure from Fatigue RelatedMicrodamage. A collaborative project between Michigan University, Monash University and Optiscan Pty Ltd.Courtesy of Professor Mark Banaszak Holl - Engineering Department, Monash University

Scale bar = 100um


7. 自体荧光成像.jpg


5、Lung endothelium triplel abelling

Contrast agent:

Lung endothelial cell nuclei: acridine orange, 0.05% i.v. (single arrow)Plasma: FITC-Dextran 70kDa, i.v. (double arrow)Distal epithelial membranes: FITC-R.Communis lectin (triple arrow)Scale bar = 100pm


9. 肺内皮三重标记.jpg


6、Gastrointestinal microflora

Helicobacter in dog stomach.


11. 胃肠道微生物成像.jpg


7、Zebrafish eye

Left: 180um Z-stack (Video) of Zebrafish eye expressing expressing endothelium specific gfp expression. Acriflavine wasused for background contrast.

Right: Maximum brightness projection of the Z-stack. Depth-colour coded in lmage ] Scale bar = 100um



左:成像深度:180μm,表达GFP的斑马鱼血管内皮成像,吖啶黄用作背景造影;

右:最明亮的一张,在Image J里深度颜色编码。Scale bar=100μm。



8、Brain cancer

Tumour island surrounded by leaky blood vessels in live rat brain. The tumour is a Glioblastoma which is highlyinfiltrative and causes hemorrhage. The contrast agent used here is intravenous fluorescein.Courtesy of researchers in Barrow Neurological institute, Phoenix, Arizona, USAScale bar = 100um



Video

Left: 400 um z stack with 4um step size Right: Maximum brightness projection Human Dermal Fibroblasts in collagen gel Alexa 488 Phalloidin (F-actin stain for cell cytoskeleton). Scale bar = 100um
Left: 250um Z-stack of zebrafish blood vessel endothelium expressing GFP Right: Maximum brightness projectionscale bar = 100um
Left: 180um Z-stack (Video) of Zebrafish eye expressing expressing endothelium specific gfp expression. Acriflavine wasused for background contrast.
Tumour island surrounded by leaky blood vessels in live rat brain. The tumour is a Glioblastoma which is highlyinfiltrative and causes hemorrhage. The contrast agent used here is intravenous fluorescein.Courtesy of researchers in Barrow Neurological institute, Phoenix, Arizona, USAScale bar = 100um
Live calcium imaging on mouse intestine with calcium sensitive stain Fluo-8. calcium level changes in isolated mouse 'Gut Wall' after nerve stimulation. lmages courtesy of Daniel Poole Monash Institute of Pharmaceutical Sciences scale bar = 100um

Tissue culture

GFP endothelium

Zebrafish eye

Brain cancer

Calcium imaging

Case

Message

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